HERPES ZOSTER mRNA VACCINE, PREPARATION METHOD THEREFOR, AND USE THEREOF

Abstract

To provide herpes zoster mRNA vaccine, a preparation method therefor, and use thereof.SOLUTION: Provided is a herpes zoster mRNA vaccine, comprising an RNA encoding a varicella-zoster virus gE glycoprotein or a variant thereof, wherein the vaccine can prevent herpes zoster infection and its complications.SELECTED DRAWING: Figure 2

Description

[Technical Field]

[0001] The present invention relates to the technical field of mRNA vaccines, and in particular to varicella-zoster mRNA vaccines and their preparation methods and uses. [Background technology]

[0002] Shingles is an infectious skin disease caused by the reactivation of the Varicella-Zoster Virus (VZV), which has long been latent in the dorsal root ganglia or cranial ganglia of the spinal cord. Shingles is a common skin disease that often causes neuropathic pain in addition to skin damage. The disease is more likely to occur in elderly people and those with immunosuppression or deficiency, and occurs most frequently in the spring and fall. The incidence rate increases significantly with age.

[0003] After the human body recovers from chickenpox infection, VZV is not completely eliminated and remains latent in cranial ganglia, dorsal root ganglia, and autonomic ganglia throughout the nervous system, where it persists for life. After VZV infection and chickenpox development, the body develops specific antibodies against VZV and cell-mediated immunity mediated by T cells (e.g., CD4 + / CD8 + These two immune functions play an important role in maintaining latent VZV infection and preventing shingles. With age or external or internal factors that inhibit the body's immune system, VZV can spontaneously reactivate and cause shingles (herpes zoster, or HZ).

[0004] Common activating factors of shingles include age factors (e.g., aging), cellular immune deficiency, genetic susceptibility, trauma, systemic diseases (e.g., diabetes, kidney disease, etc.), excessive mental stress, and fatigue. Unlike chickenpox infection in children, shingles in adults (elderly people) is prone to leaving sequelae, the most common of which is postherpetic neuralgia. It is a condition called shingles (neuralgia, PHN), and the pain can last for 3 to 12 months after the shingles has healed.

[0005] Shingles and postherpetic neuralgia are both common and frequently occurring diseases. Chronic pain caused by shingles (postherpetic neuralgia) and other complications, commonly referred to as "dragon around the waist," "snake around the waist," or "raw snake," have a serious impact on patients' quality of life, and elderly people and those with weakened immune systems are often affected by these complications. Postherpetic neuralgia remains one of the most intractable pains due to the unclear mechanism of pain.

[0006] Shingles has two distinctive features: a high infection rate and the excruciating pain it requires patients to endure. According to European and American survey data, the annual incidence of shingles is 2-6‰. The incidence rate tends to increase with age. According to statistical surveys, the incidence rate for people aged 10-49 is approximately 4‰, while the incidence rate for people aged 75 and over is as high as 14‰. Furthermore, women are more susceptible to shingles than men. Currently, the incidence of shingles is on the rise worldwide. A survey from 1994 to 2018 showed that the annual incidence of shingles increased by 3.1% each year, with a particularly significant increase in the 20-49 age group.

[0007] China does not include shingles in the range of notifiable infectious diseases of Class A and Class B, and there are relatively few epidemiological studies targeting the entire population. According to statistics, approximately 99.5% of adults over the age of 50 in China have the VZV virus harbored in their bodies, of which approximately 1.56 million new cases are infected each year. The average annual incidence rate of shingles among people aged 50-60 in China is 2.66‰, and among people over 80, The average annual incidence rate is 8.55‰. The incidence of postherpetic neuralgia (PHN) also increases with age. Studies have shown that the incidence of PHN in elderly Chinese patients with shingles is 18.8%, and in patients over 75 years of age, it is 31.7%.

[0008] Vaccines are the most effective way to prevent shingles. The main technological routes for research and development of shingles vaccines include live attenuated vaccines, recombinant protein vaccines, adenovirus vector vaccines, and mRNA vaccines.

[0009] Currently, there are only four vaccines available worldwide for preventing shingles: Zostavax®, a live attenuated vaccine from Merck & Co., Inc. (preparation discontinued), Shingrix®, a recombinant vaccine from GlaxoSmithKline (GSK), SkyZoster®, a vaccine from SK Chemical Co., Ltd. (sold only in South Korea), and a live attenuated vaccine recently launched by China's Changchun BCHT Biotechnology Co., Ltd. Merck & Co., Inc.'s live attenuated vaccine, Zostavax®, was the world's first approved shingles vaccine and was released in the United States and Europe in 2006. However, due to its low preventive efficacy (effectively reducing herpes outbreaks by approximately 70%), it is not suitable for use in people with immunosuppression or immune system disorders, significantly limiting its applicability, and production was discontinued in 2018. GlaxoSmithKline (GSK)'s recombinant protein vaccine, Shingrix®, was launched in the United States in 2017, in Europe in 2018, and filed for listing in China in 2019. Global sales of this vaccine are expected to reach $2.4 billion in 2021 and $3.2 billion in 2022, making it one of the world's top 10 best-selling vaccines for several years in a row. However, this vaccine is a protein subunit vaccine and uses GSK's specialized adjuvant, making it expensive and difficult to scale up production. More importantly, this vaccine has serious side effects, with a significantly higher incidence than live-attenuated vaccines.

[0010] mRNA vaccines are a third-generation vaccine technology following traditional vaccines and protein subunit vaccines. After invading human cells via a specific delivery system, mRNA vaccines use the body's own cells to translate mRNA into proteins. These proteins are expressed as specific viral antigens, which are recognized as foreign antigens by antigen-presenting cells (APCs), promoting the maturation of dendritic cells (DCs). These then activate B and T cells to generate a strong immune response, triggering both humoral and cellular immune responses. mRNA vaccines break the traditional vaccine immune activation model and innovatively use the body's own cells to produce antigens, thereby activating bispecific immunity, forming immunological memory, and providing longer-lasting specific immunity. The greatest advantage of mRNA vaccines is that they can be developed rapidly once the pathogen's antigen gene sequence is known. Due to its technical advantages such as rapid design and construction, high adaptability to viral mutations, and efficient universal fully synthetic production process platform and ease of standardized production, mRNA vaccines have short production links and R&D cycles, and the process is relatively simple, allowing them to be mass-produced quickly.

[0011] Compared with conventional vaccines, shingles mRNA vaccines have the following advantages: 1) They are non-infectious and non-integrating reagents, eliminating the risk of infection and insertional mutation. 2) mRNA vaccines can activate the body's specific immunity by expressing antigens in the body, resulting in more durable and effective specific immunity. 3) mRNA vaccines can be stably delivered into cells and efficiently expressed in the body. 4) The preparation of mRNA vaccines can undergo strict quality control.

[0012] gE is the main target protein in research and development of varicella zoster vaccines, and is the most abundant protein in VZV. gE is one of the most abundant glycoproteins and plays an important role in viral replication and spread between ganglion cells. As a highly glycosylated type I membrane protein, gE is transported between the endoplasmic reticulum (ER), the trans-Golgi network (TGN), and the endoplast.

[0013] In light of the above issues, in order to further reduce the risk of developing shingles in people with weakened immune systems, it is urgent to provide a new mRNA vaccine for preventing VZV that can significantly improve the effectiveness of humoral and cellular immunity after vaccination. Summary of the Invention

[0014] The present invention provides a shingles mRNA vaccine that can safely guide the body's cellular machinery to produce virtually any protein of interest, from naturally occurring proteins to antibodies and other entirely novel proteins that may have protective activity both inside and outside of cells. The shingles mRNA vaccine of the present disclosure can be used to induce a balanced immune response to VZV, including cellular and humoral immunity, but without many of the risks associated with attenuated virus vaccination.

[0015] RNA (e.g., mRNA) vaccines can be used in a variety of settings, depending on the prevalence of infectious diseases or the extent or level of unmet medical need. RNA (e.g., mRNA) vaccines can be used to protect against various genotypes, strains, and isolates of VZV. The advantage of RNA (e.g., mRNA) vaccines is that they generate much greater antibody titers and generate a response more quickly than commercially available antiviral treatments. Without being bound by theory, it is believed that, like mRNA polynucleotides, RNA vaccines are better designed to produce the proper protein conformation through translation, as they direct the natural cellular machinery. Unlike traditional vaccines, which are prepared outside the body and can potentially trigger adverse cellular responses, RNA (e.g., mRNA) vaccines are delivered to cell lines in a more natural way.

[0016] Based on the important structure and biological functions of the VZV gE protein, the present invention innovatively designed a series of mRNA vaccine candidate antigens, and compared their immunogenicity with existing mRNA vaccines (e.g., antigen YK-VZV-007, see CN108472309A; and the commercially available recombinant subunit vaccine Shingrix®). Several varicella-zoster mRNA vaccine candidates with novel antigen structures and excellent immunogenicity were successfully screened, which laid the foundation for the successful development of a varicella-zoster mRNA vaccine.

[0017] The present application provides a novel VZV-preventing mRNA vaccine by combining wild-type VZV gE glycoprotein in a specific manner (including sequence truncation, site mutation, and / or sequence deletion) to obtain various ribonucleic acids encoding VZV gE glycoprotein variants, and accordingly provides a novel VZV-preventing mRNA vaccine. Experiments have demonstrated that after vaccination, the effects of humoral immunity and cellular immunity can be significantly improved, including an increase in the content of anti-VZV gE protein IgG antibodies, immune cell (IFN-γ) levels, and the like. + and / or IL-2 + CD4 secretes + and CD8 + Increased numbers of immune cell cytokines (including IFN-γ+ and IL-2 + The present invention provides a varicella zoster mRNA vaccine that has the effect of promoting the secretion of varicella zoster (including varicella zoster).

[0018] The mRNA vaccines provided herein may comprise an RNA polynucleotide of at least one VZV glycoprotein provided by the accompanying sequence listing, or a fragment, homologue (e.g., having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity), variant or derivative thereof.

[0019] In some embodiments, the antigen encodes a VZV gE polypeptide.

[0020] In some embodiments, the VZV gE polypeptide (full length) comprises amino acids 1-623 (e.g., SEQ ID NOs: 3, 51, 55, 171, 135, 139, 143, 147, 19, 23, 151, 27, 31, 35, 39, 43, 47, 163, 167).

[0021] In some embodiments, the VZV gE polypeptide is a mutant. In some embodiments, the VZV gE mutant is a truncated polypeptide that lacks the anchor domain (ER retention domain).

[0022] In some embodiments, the truncated VZV gE polypeptide comprises amino acids 1 to 539 (eg, SEQ ID NO: 7).

[0023] In some embodiments, the truncated VZV gE polypeptide comprises amino acids 1-573 (e.g., SEQ ID NOs: 15, 59, 63, 67, 71, 75, 79, 83, 87, 91).

[0024] In some embodiments, the truncated VZV gE polypeptide comprises amino acids 1-568 (eg, SEQ ID NO: 11).

[0025] In some embodiments, the truncated VZV gE polypeptide comprises amino acids 1-587 (e.g., SEQ ID NOs: 95, 99, 103, 107, 111, 115, 119, 123, 127, 131).

[0026] In some embodiments, the truncated VZV gE polypeptide comprises amino acids 1-601 (eg, SEQ ID NOs: 155, 159).

[0027] In some embodiments, the VZV gE mutant is a full-length or truncated polypeptide. It also includes mutations in one or more Golgi or trans-Golgi network (TGN) targeting motifs, which decrease VZV gE polypeptide targeting or localization to the Golgi or TGN. In particular, the inventors have identified motifs that direct gE protein localization to the Golgi or trans-Golgi network as "A 568 Y 569 R 570 V 571 " motif (specific mutant antigen sequences thereof include, for example, YK-VZV-011 (SEQ ID NO: 35) and, in particular, the A568D and / or Y569K mutations).

[0028] Additionally, in some embodiments, the VZV gE mutant may have a Y582A / G mutation, or a Y569K / A mutation, or a combination of Y582G and Y569A mutations. 593 EST 596 DT 598 (SEQ ID NO: 182) Also included are phosphorylated acidic motifs such as site mutations.

[0029] In some embodiments, the mutant VZV gE polypeptide is a full-length or truncated polypeptide, which contains one or more VZV gE internalization or endocytosis motifs Y, ... 582 AGL 585Also included is a mutation in (SEQ ID NO: 185), which reduces endocytosis of the VZV gE polypeptide.

[0030] In some embodiments, the VZV gE mutant is a VZV gE mutant having a Y569K mutation. long polypeptides (SEQ ID NOs: 27, 31, 35, 39, 43, 47, 143, 147).

[0031] In some embodiments, the VZV gE mutant is a full-length polypeptide having a Y582A mutation (SEQ ID NOs: 23, 27, 31, 43, 47, 135, 139, 171).

[0032] In some embodiments, the VZV gE mutant is a full-length polypeptide having Y582A and Y569K mutations (SEQ ID NOs: 27, 31, 43, 47).

[0033] In some embodiments, the VZV gE mutant is A 593 EA 595 A 596 DA 598 (SEQ ID NO: 183).

[0034] In some embodiments, the VZV gE mutant is A 593 EA 595 A 596 DA 598 (SEQ ID NO: 183) and a Y582G mutation, or a Y569K mutation, or a combined Y582G and Y569K mutation.

[0035] In some embodiments, the VZV gE mutant comprises one or more site mutations selected from the group consisting of A568D, Y569K, Y569A, R570E, V571K, Y582A, S593A, S595A, T596A, and T598A, preferably A568D, Y569K, R570E, and V571K.

[0036] In some embodiments, the site mutation is any one of the following groups: 1)A568D, Y569K, R570E, V571K, (YK-VZV-023, YK-VZV-011) 2)A568D, Y569A, R570E, V571K, (YK-VZV-024) 3)Y569K, R570E, V571K, (YK-VZV-025) 4)A568D, Y569K, R570E, V571K, Y582A, (YK-VZV-030, YK-VZV-013) 5)A568D, Y569K, R570E, V571K, Y582G, (YK-VZV-031, YK-VZV-038) 6)A568D, Y582A, (YK-VZV-032) 7)Y569K, Y582A, (YK-VZV-033, YK-VZV-009) 8)R570E, Y582A, (YK-VZV-034) 9)V571K, Y582A, (YK-VZV-035) 10)S593A, S595A, T596A, T598A, (YK-VZV-016, YK-VZV-044) 11)Y582A, S593A, S595A, T596A, T598A, (YK-VZV-037) 12)A568D, Y569K, R570E, V571K, Y582G, S593A, S595A, T596A, T598A, (YK-VZV-039) 13)A568D, Y569K, R570E, V571K, Y582A, S593A, S595A, T596A, T598A, (YK-VZV-014) 14)Y582G, S593A, S595A, T596A, T598A, (YK-VZV-040) 15)Y569K, Y582A, S593A, S595A, T596A, T598A, (YK-VZV-010) 16)A568D, Y569K, R570E, V571K, S593A, S595A, T596A, T598A, (YK-VZV-012) 17) Y582G, (YK-VZV-029) 18) A568D, (YK-VZV-018) 19) Y569K, (YK-VZV-020) 20) R570E, (YK-VZV-021) 21) V571K, (YK-VZV-022) 22)Y582A, (YK-VZV-028, YK-VZV-036)

[0037] In some embodiments, the VZV gE mutant further comprises a sequence deletion, wherein the sequence deletion is selected from any one or more of the following group: 1) A 568 , 2) Y 569 RVDKSPYNQS 579 , 3) Y 569 RVDKSPYNQSMYYAGLPV 587 .

[0038] The VZV RNA antigen of the present disclosure has multiple combination types of mutations, including C-terminal amino acid truncation of the gE antigen, and deletion or substitution of key amino acid sites, which will be further described below through specific examples.

[0039] The present invention relates to a method for producing a VZV vector comprising the steps of: providing a template capable of being transcribed into VZV RNA; Further provided is a method for preparing said composition, comprising the step of transcribing using said template under conditions suitable for transcription into said RNA.

[0040] In some embodiments, the preparation method comprises: gene synthesis of the DNA sequence of the VZV gE antigen followed by homologous recombination into the PVAX1 vector; Obtaining the plasmid after transformation and cultivation of E. coli; obtaining a linearized plasmid after enzymatic digestion of the plasmid; The method includes the steps of using T7 RNA polymerase to generate a linearized plasmid as a template, inputting NTP raw materials, co-transcription and capping, and then purifying the resulting RNA, preferably mRNA.

[0041] In some embodiments, the purification method comprises one or more of lithium chloride precipitation, affinity chromatography, solution exchange by ultrafiltration, and cellulose chromatography.

[0042] In some embodiments, the method for preparing the composition includes the steps of synthesizing a DNA sequence based on an antigen gene and inserting it into a plasmid DNA construct; cleaving the plasmid DNA with a DNA restriction endonuclease to form a linearized template; using the linearized plasmid as a template under the catalytic action of T7 RNA polymerase, adding key raw materials such as NTPs and cap analogs, and transcribing and preparing mRNA to modify the RNA; adding a cap to the 5' end; and removing contaminants, enzymes, free nucleotides, and other impurities from the modified mRNA by methods such as lithium chloride precipitation and affinity chromatography.

[0043] In a third aspect, the composition of the present invention is in the form of a vaccine, ie, a varicella zoster vaccine, which further comprises a pharmaceutically acceptable carrier in addition to the composition.

[0044] In some preferred embodiments, the pharmaceutically acceptable carrier comprises a lipid mixture, preferably a lipid nanoparticle LNP.

[0045] In some preferred embodiments, the lipid nanoparticles may comprise, for example, cationic lipids. , neutral lipids, structured lipids and polymer-conjugated lipids.

[0046] In a fourth aspect, the present invention further provides a method for preparing a vaccine composition, comprising mixing the VZV RNA with the pharmaceutically acceptable carrier, e.g., encapsulating at least a portion of the RNA in lipid nanoparticles. In some embodiments, the method comprises mixing RNA, preferably mRNA, with lipid nanoparticles, encapsulating them in lipid nanoparticles, and purifying to remove unencapsulated components.

[0047] In some more specific embodiments, the preparation method specifically includes the steps of incorporating the mRNA into the lipid nanoparticles (LNPs), i.e., mixing a mixture of purified mRNA and lipids in a microfluidic chip and allowing them to self-assemble to form lipid nanoparticles; encapsulating the mRNA in the lipid nanoparticles; dialyzing or filtering the lipid nanoparticle solution to remove unencapsulated mRNA, non-aqueous solvents, and bacteria; storing the filtered lipid nanoparticle solution and packaging it into sterile vials, thereby completing the preparation and production of the mRNA vaccine.

[0048] The present application first discovered the following:

[0049] The present application provides a novel mRNA vaccine for preventing VZV infection, which is based on wild-type VZV gE glycoprotein and is combined in a specific manner (including sequence truncation, site mutation, and / or sequence deletion) to obtain several ribonucleic acids encoding VZV gE glycoprotein variants. Experiments have demonstrated that after vaccination, the effects of humoral immunity and cellular immunity can be significantly improved, including an increase in the content of anti-VZV gE protein IgG antibodies, immune cell (IFN-γ) levels, and the like. + and / or IL-2 + CD4 secretes + and CD8 + Increased numbers of immune cell cytokines (including IFN-γ + and IL-2 +The present application provides a varicella zoster mRNA vaccine that has the effect of promoting the secretion of varicella zoster (including varicella zoster). Specifically, the present application includes the following discoveries and results.

[0050] 1. 43 VZVs designed Western blot detection was performed on the gE antigen variants, and the results showed that all 39 antigen sequences had significant protein expression, including YK-VZV-001, YK-VZV-003, YK-VZV-009, YK-VZV-010, YK-VZV-011, YK-VZV-012, YK-VZV-013, YK-VZV-014, YK-VZV-015, YK-VZV-016, YK-VZV-017, YK-VZV-018, YK-VZV-019, YK-VZV-020, YK-VZV-021, YK-VZV-022, YK-VZV-023, YK-VZV-024, YK-VZV-025, and YK-VZV-026. 26, YK-VZV-027, YK-VZV-028, YK-VZV-029, YK-VZV-030, YK-VZV-031, YK-VZV-032, Y K-VZV-033, YK-VZV-034, YK-VZV-035, YK-VZV-036, YK-VZV-037, YK-VZV-038, YK-VZ These include V-039, YK-VZV-040, YK-VZV-044, and YK-VZV-004 (Comparative Example 1), YK-VZV-006 (Comparative Example 2), YK-VZV-045 (Comparative Example 3), and YK-VZV-007 (Comparative Example 4), which can be used as effective candidate antigens for the next step of immunogenicity screening and evaluation.

[0051] 2. Based on the above 39 sequences, the gE-specific antibody IgG titer results on day 42 identified the following 14 variants: YK-VZV-009, YK-VZV-011, YK-VZV-013, YK-VZV-014, YK-VZV-010, YK-VZV-012, YK-VZV-018, YK-VZV-020, YK-VZV-030, YK-VZV-031, YK-VZV-038, YK-VZV-021, YK-VZV-024, YK-VZV-026, YK-VZV-028, YK-VZV-029, YK-VZV-030, YK-VZV-031, YK-VZV-038, YK-VZV-032, YK-VZV-033, YK-VZV-034, YK-VZV-035, YK-VZV-036, YK-VZV-037, YK-VZV-039, YK-VZV-040, YK-VZV-041, YK-VZV-042, YK-VZV-043, YK-VZV-044, YK-VZV-046, YK-VZV-047, YK-VZV-049, YK-VZV-050, YK-VZV-051, YK-VZV-052, YK-VZV-053, YK-VZV-054, YK-VZV-055, YK-VZV-056, YK-VZV-057, YK-VZV-058, Y Extract V-028 and four control sequences: YK-VZV-004 (Comparative Example 1), YK-VZV-006 (Comparative Example 2), YK-VZV-045 (Comparative Example 3), and YK-VZV-007 (Comparative Example 4).

[0052] The above sequences are 3.3 to 5.3 times more potent than the Shingrix® positive control and 1.8 to 2.8 times more potent than the YK-VZV-007 (Comparative Example 4) sequence. All of these sequences are superior to Shingrix®, YK-VZV-007 (Comparative Example 4), YK-VZV-004 (Comparative Example 1), YK-VZV-006 (Comparative Example 2), and YK-VZV-045 (Comparative Example 3).

[0053] Mouse spleen secretes IFN-γ, IL-2, and CD4 + T cells and CD8 + The results of measuring the proportion of T cells reveal the following:

[0054] (1) CD4 + According to the proportion of T cells, YK-VZV-010, YK-VZV-013, YK-VZV-011, YK-VZV-020, YK-VZV-012, YK-VZV-018, YK-VZV-028, YK-VZV-038, and YK-VZV-031 were screened, and the above nine sequences were IFN-γ + CD4 + T cells and IL-2 + CD4 + The total percentage of T cells was more than 2-fold higher than the Shingrix® positive control, up to 7-fold higher, and approximately 1.8-5.3-fold higher than YK-VZV-007 (Comparative Example 4). IFN-γ + CD4 + The percentage of T cells was 1.00 to 2.00%, which was more than three times higher than the Shingrix (registered trademark) positive control, up to 6.7 times higher, and approximately 2.2 to 4.4 times higher than YK-VZV-007 (Comparative Example 4). IL-2 + CD4 +The percentage of T cells exceeded 1% in both cases, which was 3.7 times higher than that of the Shingrix® positive control vaccine and approximately 2.2 times higher than that of YK-VZV-007.

[0055] (2) Furthermore, CD8 + Five sequences, YK-VZV-010, YK-VZV-013, YK-VZV-020, YK-VZV-018, and YK-VZV-011, were screened according to the proportion of T cells. IFN-γ + CD8 + T cells and IL-2 + CD8 + The total percentage of T cells was 4 to 7 times higher than that of the Shingrix® vaccine and approximately 3 to 5 times higher than that of YK-VZV-007 (Comparative Example 4). IFN-γ + CD8 + The proportion of T cells was optimal, being 5.0% or more in all cases, 4.5 to 7.0 times higher than that of the Shingrix® positive control vaccine, and approximately 2.5 to 4.5 times higher than that of YK-VZV-007. IL-2 + CD8 + The proportion of T cells was optimal, reaching 0.3-0.5%, which was 2.5-4.0 times higher than that of the Shingrix® positive control vaccine and approximately 1.5-3.0 times higher than that of YK-VZV-007. All of these were significantly higher than those of the comparative examples (YK-VZV-004, YK-VZV-006, YK-VZV-045, and YK-VZV-007).

[0056] 4. To verify the above results, the VZV-binding-gE antibody IgG titers in the serum of immunized mice on day 56 were further measured and verified based on the results measured on day 42.

[0057] The results showed that 56 days after immunization of mice with the YK-VZV-009, YK-VZV-010, YK-VZV-011, YK-VZV-013, YK-VZV-020, YK-VZV-014, YK-VZV-012, YK-VZV-018, YK-VZV-021, YK-VZV-024, YK-VZV-028, YK-VZV-030, YK-VZV-031, and YK-VZV-038 vaccines, the gE-specific antibody IgG of the above mRNA vaccines was significantly higher than that of the control group. All titers were superior to the Shingrix® positive control, and were approximately 1.8 to 5.0 times stronger than the Shingrix® vaccine.

[0058] Among them, the antibody GMTs of YK-VZV-011, YK-VZV-013, YK-VZV-020, YK-VZV-018, and YK-VZV-010 were 650 × 10 4 or more, which was 4.0 to 5.0 times or more that of the Shingrix (registered trademark) vaccine and more than twice that of YK-VZV-007 (Comparative Example 4). Furthermore, both were significantly superior to YK-VZV-004 (Comparative Example 1) and YK-VZV-045 (Comparative Example 3).

[0059] 5. Combining the results of IgG and multiparameter flow cytometry detection, five optimal antigen sequences (including YK-VZV-010, YK-VZV-013, YK-VZV-011, YK-VZV-020, and YK-VZV-018) were screened and further verified using the ELISpot method. The results are as follows:

[0060] (1) The amount of cytokine IFN-γ stimulated by the mRNA vaccines to secrete from T cells exceeded 500 SFU in all cases, with the highest reaching nearly 600 SFU, which was approximately 16 to 19 times that of the Shingrix® positive control and 2.5 to 3.0 times that of Comparative Example 4 (YK-VZV-007), and was significantly superior to the cytokine secretion stimulated by the corresponding mRNA vaccines in Comparative Example 1 (YK-VZV-004), Comparative Example 2 (YK-VZV-006), or Comparative Example 3 (YK-VZV-045).

[0061] Among them, the mRNA vaccine prepared with the YK-VZV-010 sequence reached a maximum of 598.8 SFU, 18.7 times higher than the Shingrix® positive control and nearly three times higher than the YK-VZV-007 control.

[0062] The IFN-γ secretion levels, from highest to lowest, were YK-VZV-010, YK-VZV-011, YK-VZV-018, YK-VZV-020, and YK-VZV-013, with no significant difference between the groups, and the immune effects were all good and significant.

[0063] (2) The amount of cytokine IL-2 secreted by T cells stimulated by the mRNA vaccines was 400 to 600 SFU or more in all cases, which was 10 to 16 times that of the Shingrix (registered trademark) positive control and 2.0 to 3.0 times that of the YK-VZV-007 control, and was significantly superior to the cytokine secretion stimulated by the mRNA vaccines corresponding to Comparative Example 1 (YK-VZV-004), Comparative Example 2 (YK-VZV-006), or Comparative Example 3 (YK-VZV-045).

[0064] Among them, the mRNA vaccine prepared with the YK-VZV-010 sequence reached a maximum of 614.5 SFU, 16.0 times that of the Shingrix® positive control and more than three times that of the YK-VZV-007 control.

[0065] The IL-2 secretion levels, from highest to lowest, were YK-VZV-010, YK-VZV-011, YK-VZV-018, YK-VZV-013, and YK-VZV-020, and the differences between the groups were not significant. The immune effects were all good and significant.

[0066] 6. Furthermore, the antigen localization results of the above five sequences indicate that they are localized in the Golgi apparatus, the cell membrane, or the cytoplasm. [Brief explanation of the drawings]

[0067] [Figure 1]It is a deletion mutant of the major functional domain and sequence of the VZV gE protein. [Figure 2] YK-VZV-001 to YK-VZV-045 antigen expression. [Figure 3] Cellular localization of VZV gE antigen in YK-VZV-010, YK-VZV-011, and YK-VZV-013. [Figure 4] Cellular localization of VZV gE antigen in YK-VZV-018 and YK-VZV-020. Here, green represents the gE antigen, yellow and red represent GM130 and TGN46, respectively. The white arrowhead represents the plasma membrane, and the orange arrowhead represents the Golgi apparatus. DETAILED DESCRIPTION OF THE INVENTION

[0068] The present invention will be further described below by way of examples. It should be understood that the examples of the present invention are only used to illustrate the present invention and do not limit the present invention, and any simple improvements of the present invention based on the technical solutions of the present invention are within the protection scope of the present invention.

[0069] As used herein, the term "selected from" refers to selecting one, or any combination of two, three, or more, from the listed objects or elements.

[0070] It is understood that the term "particular compound" as used herein may include said compound, its N-oxide, its solvate, its pharmaceutically acceptable salt, its stereoisomer, and mixtures thereof.

[0071] Designing antigen sequences: An antigen is a protein capable of inducing an immune response (e.g., causing the immune system to produce antibodies against the antigen). As used herein, unless otherwise specified, the term "antigen" covers immunogenic proteins and immunogenic fragments that induce or are capable of inducing an immune response against varicella-zoster virus (e.g., VZV). It is understood that the term "protein" covers peptides, and the term "antigen" covers antigenic fragments. Table 1 provides exemplary sequences of VZV antigens and RNAs encoding the VZV antigens of the compositions of the present disclosure.

[0072] Nucleic Acid: A nucleic acid comprises a polymer of nucleotides (nucleotide monomers). The compositions of the present disclosure comprise RNA having an open reading frame (ORF) encoding a VZV antigen. In some embodiments, the RNA is messenger RNA (mRNA). In some embodiments, the RNA (e.g., mRNA) further comprises a 5' UTR, a 3' UTR, a poly(A) tail, and / or a 5' cap analog.

[0073] Messenger RNA (mRNA): Any RNA that encodes (at least one) protein (naturally occurring, non-naturally occurring, or modified amino acid polymer) and that is translated in vitro, in vivo, in situ, or ex vivo to encode the protein. Unless otherwise specified, the nucleic acid sequences described in this application may recite a "T" in a representative DNA sequence, provided that one of skill in the art will understand that when the sequence represents RNA (e.g., mRNA), the "T" is replaced with a "U." Thus, any DNA disclosed and identified herein by a particular sequence identification number also discloses the corresponding RNA (e.g., mRNA) sequence complementary to that DNA, in which each "T" in the DNA sequence is replaced with a "U."

[0074] Open Reading Frame (ORF): A continuous DNA or RNA sequence that begins with a start codon (e.g., methionine (ATG)) and ends with a stop codon (e.g., TAA, TAG, or TGA, or UAA, UAG). ORFs typically encode proteins. The sequences disclosed herein may also include additional elements such as 5' and 3' UTRs.

[0075] Variant RNA:Antigen variant or other polypeptide variant refers to a molecule whose amino acid sequence differs from the wild-type, native or reference sequence. An antigen / polypeptide variant is a molecule whose amino acid sequence differs from the native sequence or The variants may have substitutions, deletions, and / or insertions at specific positions within the amino acid sequence compared to the reference sequence. Typically, the variants have at least 50% identity with the wild-type, native, or reference sequence. In some embodiments, the variants have at least 80% or at least 90% identity with the wild-type, native, or reference sequence.

[0076] In some embodiments, the compositions comprise RNA or RNA ORFs and comprise the nucleotide sequence of any of the sequences provided herein (see, e.g., Sequence Listing and Table 1), or a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% or 100% identity to the nucleotide sequence of any of the sequences provided herein.

[0077] The term "identity" refers to the relationship between two or more polypeptide (e.g., antigen) or polynucleotide (nucleic acid) sequences, as determined by comparing the sequences. Identity also refers to the degree of sequence relatedness between or within sequences, as determined, for example, by the number of matches between two or more strings of amino acid or nucleic acid residues. Identity measures the percentage of identical matches between the smaller of two or more sequences with gapped alignments (if any) processed by a particular mathematical model or computer program (e.g., an "algorithm"). The identity of related antigens or nucleic acids can be readily calculated using known methods. "Percent identity" when applied to polypeptide or polynucleotide sequences is defined as the percentage of residues (amino acid residues or nucleic acid residues) in a candidate amino acid or nucleic acid sequence that are identical to the residues in the amino acid or nucleic acid sequence of a second sequence, after aligning the sequences and introducing gaps (if necessary) to achieve the maximum percent identity. Methods and computer programs for comparison are well known in the art. While identity is dependent on the calculation of percent identity, it is understood that the introduction of gaps and penalties in the calculation may result in different identity values. Generally, variants of a particular polynucleotide or polypeptide (e.g., antigen) will have at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, but less than 100%, sequence identity with the particular reference polynucleotide or polypeptide, as determined by sequence alignment programs and parameters described herein and known to those of skill in the art. Tools for such comparisons include the BLAST suite (Stephen F. Altschul et al. (1997), "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs," Nucleic Acids Res. 25:3389-3402) and the like.

[0078] Therefore, any polynucleotide encoding a peptide or polypeptide having substitutions, insertions, and / or additions or deletions relative to a reference sequence, particularly the antigen sequences disclosed herein, is within the scope of the present disclosure. Amino acid residues located in the carboxyl and amino terminal regions of the amino acid sequence of a peptide or protein may optionally be deleted to provide a truncated sequence. Based on the structure and use of the sequence, the gE protein has several important domains related to its main functions (Figure 1). The gE protein has several important domains related to its main functions (Figure 3). (1) TM transmembrane domain, consisting of amino acids 539 to 559; (2) A568YRV571: mediates gE transport between the endoplasmic reticulum (ER), the trans-Golgi network (TGN), and the endoplasm; (3) Y582AGL585: mediates gE transport between the endoplasmic reticulum (ER), the trans-Golgi network (TGN), and the endoplasm; (4) S593ES595T596DT598: important glycosylation site, mediates gE transport between the endoplasmic reticulum (ER), the trans-Golgi network (TGN), and the endoplasm. In some embodiments, important structural sites are excluded. Sequences can be removed and substituted with appropriate residues to alter antigen transport and expression, thereby enhancing the immunogenicity of the antigen. Such sequences can be readily identified by one of skill in the art. It is also understood that some sequences provided herein contain sequence tags or terminal peptide sequences (e.g., N- or C-termini) that may be deleted prior to use in, for example, preparing an RNA (e.g., mRNA) vaccine.

[0079] As will be recognized by those skilled in the art, protein fragments, functional protein domains, and homologous proteins are also considered to be within the scope of the target Varicella-Zoster virus antigen. For example, the present specification provides any protein fragment (referring to a polypeptide sequence that is at least one amino acid residue shorter than the reference antigen sequence but otherwise identical) of a reference protein, provided that the fragment is immunogenic and confers a protective immune response against Varicella-Zoster virus. In addition to identical but truncated variants of the reference protein, in some embodiments, the antigen contains 2, 3, 4, 5, 6, 7, 8, 9, 10, or more site mutations as shown in any of the sequences provided or referenced herein. The length of the antigen / antigenic polypeptide can range from about 4, 6, or 8 amino acids to the full-length protein.

[0080] VZV Antigen Mutants: The present invention encompasses mutant VZV antigenic polypeptides. In some embodiments, the mutant VZV antigenic polypeptide is a mutant VZV gE polypeptide. The mutant VZV gE polypeptide is designed to avoid ER / Golgi apparatus retention of the peptide, resulting in increased surface expression of the antigen. In some embodiments, the mutant gE polypeptide is truncated to remove the ER retention portion or cytoplasmic tail portion of the polypeptide. In some embodiments, the mutant VZV gE polypeptide is mutated to reduce localization of the VZV polypeptide to the ER / Golgi apparatus / TGN. The modification inhibits ER trapping and therefore promotes transport to the plasma membrane. Thus, in some embodiments, the VZV glycoprotein is a mutant gE polypeptide. VZV gE contains a TGN-targeting sequence at its C-terminus and is transported from the ER to the TGN in infected and gE-transfected cells. It is believed that most gE in the TGN is retrieved from the plasma membrane via endocytosis, delivered to the TGN from endosomes, and then recycled back to the plasma membrane. gE accumulates at the TGN along with other VZV proteins (e.g., envelope proteins) involved in the generation of fully enveloped VZV virions. Therefore, mutations that reduce TGN localization and endocytosis favor gE transport to the plasma membrane, and mutant VZV gE polypeptides can be any truncated polypeptide lacking the anchor domain (ER retention domain). For example, mutant VZV gE polypeptides can be truncated VZV gE polypeptides containing at least amino acids 1-124, e.g., amino acids 1-124, 1-140, 1-160, 1-200, 1-250, 1-300, 1-350, 1-360, 1-400, 1-450, 1-500, 1-511, 1-550, and 1-561, as well as polypeptide fragments having fragment sizes within the recited size ranges. In some embodiments, mutant VZV gE polypeptides are truncated polypeptides lacking the carboxy-terminal tail domain. Thus, in some embodiments, the truncated VZV gE polypeptide comprises amino acids 1-573 of SEQ ID NO:59.In some embodiments, the mutant VZV gE polypeptide has at least one mutation in one or more motifs associated with ER retention, wherein the mutation in the one or more motifs results in reduced retention of the VZV gE polypeptide in the ER and / or Golgi apparatus. In some embodiments, the mutant VZV gE polypeptide has at least one mutation in one or more phosphorylated acidic motifs. For example, the mutant VZV gE polypeptide may be a full-length VZV gE polypeptide having a Y582G mutation, a Y569K mutation, or both a Y582G mutation and a Y569K mutation. Alternatively, the mutant VZV gE polypeptide may be, for example, an antigenic VZV gE polypeptide comprising amino acids 1-573 of VZV gE and having a Y569K mutation. Alternatively, the mutant VZV gE polypeptide may be an antigenic fragment having a mutation in an acidic phosphorylation motif (e.g., an SSTT motif). For example, the mutant VZV gE polypeptide may be an antigenic fragment having the sequence AEAADA (SEQ ID NO: 183).

[0081] Naturally occurring eukaryotic mRNA molecules may contain stabilizing elements, including, but not limited to, untranslated regions (UTRs) at their 5'-end (5'UTR) and / or their 3'-end (3'UTR), in addition to other structural features, such as a 5'-cap structure or a 3'-poly(A) tail. Both the 5'UTR and the 3'UTR are typically transcribed from genomic DNA and are components of the premature mRNA. Typically, structural features characteristic of mature mRNA (e.g., a 5'-cap and a 3'-poly(A) tail) are added to the transcribed (premature) mRNA during mRNA processing.

[0082] In some embodiments, the composition comprises an RNA polynucleotide having an open reading frame encoding at least one antigenic polypeptide with at least one modification and at least one 5'-end cap, formulated within a lipid nanoparticle. 5'-capping of the polynucleotide can be accomplished simultaneously during an in vitro transcription reaction using a chemical RNA cap analog to generate a 5'-guanosine cap structure, according to the manufacturer's protocol. In some cases, 5'-capping of the modified RNA can also be accomplished post-transcriptionally using vaccinia virus capping enzyme to generate a "cap" structure. Cap 1 structure can be generated using vaccinia virus capping enzyme and 2'-O-methyltransferase to generate m7G(5')ppp(5')G-2'-O-methyl. Cap 2 structure can be generated on top of the Cap 1 structure by subsequent 2'-O-methylation of the penultimate 5' nucleotide using 2'-O-methyltransferase. The Cap 3 structure can be generated on the Cap 2 structure by subsequent 2'-O-methylation of the penultimate 5' nucleotide using a 2'-O-methyltransferase. The enzyme can be obtained from recombinant sources. The 3'-poly(A) tail is typically a series of adenine nucleotides added to the 3' end of a transcribed mRNA. In some cases, it may contain up to about 400 adenine nucleotides. In some embodiments, the length of the 3'-poly(A) tail may be a necessary factor for the stability of an individual mRNA.

[0083] Chemical Modification: In some embodiments, the compositions of the present disclosure comprise RNA having an open reading frame encoding a VZV antigen, wherein the nucleic acid comprises nucleotides and / or nucleosides that may be standard (unmodified) or modified as known in the art. In some embodiments, the nucleotides and nucleosides of the present disclosure comprise modified nucleotides or nucleosides. Such modified nucleotides and nucleosides may be naturally occurring modified nucleotides and nucleosides or non-naturally occurring modified nucleotides and nucleosides. Such modifications may include modifications in the sugar, backbone, or nucleobase moieties of the nucleotides and / or nucleosides that are well known in the art.

[0084] In some embodiments, the naturally occurring modified nucleotides or nucleosides of the present disclosure are nucleotides or nucleosides that are commonly known or known in the art. Non-limiting examples of such naturally occurring modified nucleotides and nucleosides can be found, inter alia, in the widely known MODOMICS database.

[0085] In some embodiments, the non-naturally occurring modified nucleotides or The nucleoside is a nucleotide or nucleoside commonly known or known in the art. Non-limiting examples of such non-naturally occurring modified nucleotides and nucleosides can be found, inter alia, in published U.S. patent applications PCT / US2012 / 058519, PCT / US2013 / 075177, PCT / US2014 / 058897, PCT / US2014 / 058891, PCT / US2014 / 070413, PCT / US2015 / 36773, PCT / US2015 / 36759, PCT / US2015 / 36771, or PCT / IB2017 / 051367, all of which are incorporated herein by reference.

[0086] Thus, the nucleic acids of the present disclosure (e.g., DNA nucleic acids and RNA nucleic acids such as mRNA nucleic acids) can include standard nucleotides and nucleosides, naturally occurring nucleotides and nucleosides, non-naturally occurring nucleotides and nucleosides, or any combination thereof.

[0087] In some embodiments, nucleic acids of the disclosure (e.g., DNA nucleic acids and RNA nucleic acids such as mRNA nucleic acids) contain multiple (one or more) different types of standard and / or modified nucleotides and nucleosides. In some embodiments, a particular region of a nucleic acid contains one, two, or more (optionally different) types of standard and / or modified nucleotides and nucleosides.

[0088] In some embodiments, modified RNA nucleic acids (e.g., modified mRNA nucleic acids) introduced into a cell or organism exhibit reduced degradation within the cell or organism, respectively, compared to unmodified nucleic acids comprising standard nucleotides and nucleosides.

[0089] In some embodiments, modified RNA nucleic acids (e.g., modified mRNA nucleic acids) introduced into a cell or organism may exhibit reduced immunogenicity in the cell or organism, respectively, compared to unmodified nucleic acids comprising standard nucleotides and nucleosides.

[0090] In some embodiments, nucleic acids (e.g., RNA nucleic acids, such as mRNA nucleic acids) contain non-naturally occurring modified nucleotides that are introduced during or after nucleic acid synthesis to achieve a desired function or property. Modifications may occur in the internucleotide bond, the purine or pyrimidine base, or the sugar. Modifications can be introduced at the end of the chain or any other position in the chain by chemical synthesis or using a polymerase. Any region of the nucleic acid may be chemically modified.

[0091] The present disclosure provides modified nucleosides and nucleotides of nucleic acids (e.g., RNA nucleic acids such as mRNA nucleic acids). A "nucleoside" refers to a compound comprising a sugar molecule (e.g., pentose or ribose) or a derivative thereof in combination with an organic base (e.g., a purine or pyrimidine) or a derivative thereof (also referred to herein as a "nucleobase"). A "nucleotide" refers to a nucleoside comprising a phosphate group. Modified nucleotides can be synthesized by any useful method, such as chemical, enzymatic, or recombinant methods, to include one or more modified or unnatural nucleosides. A nucleic acid may comprise one or more regions of linked nucleosides. Such regions may have variable backbone linkages. The linkages may be standard phosphodiester linkages, in which case the nucleic acid comprises a region of nucleotides.

[0092] Modified nucleotide base pairs not only cover standard adenosine-thymine, adenosine-uracil, or guanosine-cytosine base pairs, but also include base pairs formed between nucleotides and / or modified nucleotides containing non-standard or modified bases, where the arrangement of hydrogen bond donors and hydrogen bond acceptors can be different between a non-standard base and a standard base, or between two bases. Non-standard base pairs may be formed between complementary base structures, such as hydrogen bonds in nucleic acids having at least one chemical modification. An example of such a non-standard base pair is a base pair between the modified nucleotide inosine and adenine, cytosine, or uracil. Any combination of base / sugar or linker may be incorporated into the nucleic acids of the present disclosure.

[0093] In some embodiments, modified nucleobases in a nucleic acid (e.g., an RNA nucleic acid such as an mRNA nucleic acid) comprise 1-methyl-psudouridine (m1ψ), 1-ethyl-psudouridine (e1ψ), 5-methoxy-uridine (mo5U), 5-methyl-cytidine (m5C), and / or pseudouridine (ψ). In some embodiments, modified nucleobases in a nucleic acid (e.g., an RNA nucleic acid such as an mRNA nucleic acid) comprise 5-methoxymethyluridine, 5-methylthiouridine, 1-methoxymethylpsudouridine, 5-methylcytidine, and / or 5-methoxycytidine. In some embodiments, a polyribonucleotide comprises a combination of at least two (e.g., two, three, four, or more) of any of the above modified nucleobases, including, but not limited to, chemical modifications.

[0094] In some embodiments, an mRNA of the disclosure comprises 1-methyl-psudouridine (m1ψ) substitutions at one or more, or all, uridine positions in the nucleic acid.

[0095] In some embodiments, an mRNA of the disclosure comprises 1-methyl-psudouridine (m1ψ) substitutions at one or more or all uridine positions in the nucleic acid and 5-methylcytidine substitutions at one or more or all cytidine positions in the nucleic acid.

[0096] In some embodiments, an mRNA of the disclosure comprises pseudouridine (ψ) substitutions of one or more or all uridine positions of the nucleic acid.

[0097] In some embodiments, for certain modifications, the mRNA is uniformly modified (e.g., fully modified, modified throughout the entire sequence). For example, the nucleic acid is uniformly modified with 1-methyl-psudouridine, meaning that all uridine residues in the mRNA sequence are replaced with 1-methyl-psudouridine. Similarly, the nucleic acid can be uniformly modified with any type of nucleoside residue present in the sequence, substituting modified residues (e.g., those listed above).

[0098] The nucleic acid may contain from about 1% to about 100% modified nucleotides (relative to the total nucleotide content, or relative to one or more types of nucleotides, i.e., any one or more of A, G, U, or C), or any percentage therebetween (e.g., 1% to 20%, 1% to 25%, 1% to 50%, 1% to 60%, 1% to 70%, 1% to 80%, 1% to 90%, 1% to 95%, 10% to 20%, 10% to 25%, 10% to 50%, 10% to 60%, 10% to 70%, 10% to 80%, 10% to 90%, 10% to 10 ... The percentages may include 10% to 95%, 10% to 100%, 20% to 25%, 20% to 50%, 20% to 60%, 20% to 70%, 20% to 80%, 20% to 90%, 20% to 95%, 20% to 100%, 50% to 60%, 50% to 70%, 50% to 80%, 50% to 90%, 50% to 95%, 50% to 100%, 70% to 80%, 70% to 90%, 70% to 95%, 70% to 100%, 80% to 90%, 80% to 95%, 80% to 100%, 90% to 95%, 90% to 100%, and 95% to 100%). It is understood that the presence of unmodified A, G, U, or C accounts for the remaining percentage.

[0099] The mRNA may contain as little as 1%, as much as 100% modified nucleotides, or any percentage therebetween, e.g., at least 5% modified nucleotides, at least 10% modified nucleotides, at least 25% modified nucleotides, at least 50% modified nucleotides, at least 80% modified nucleotides, or at least 90% modified nucleotides. For example, the nucleic acid may contain modified nucleotides. In some embodiments, at least 5%, at least 10%, at least 25%, at least 50%, at least 80%, at least 90%, or 100% of the uracils in the nucleic acid are substituted with modified uracils (e.g., 5-substituted uracils). The modified uracils may be substituted with a compound having a single unique structure, or with multiple compounds having different structures (e.g., two, three, four, or more unique structures). In some embodiments, at least 5%, at least 10%, at least 25%, at least 50%, at least 80%, at least 90%, or 100% of the cytosines in the nucleic acid are substituted with modified cytosines (e.g., 5-substituted cytosines). The modified cytosines may be substituted with a compound having a single unique structure, or with multiple compounds having different structures (e.g., two, three, four, or more unique structures).

[0100] Untranslated Region (UTR): The mRNA of the present disclosure may contain one or more regions or portions that act or function as untranslated regions. When the mRNA is designed to encode at least one target antigen, the nucleic acid may contain one or more of these untranslated regions (UTRs). The wild-type untranslated region of a nucleic acid is transcribed but not translated. In an mRNA, the 5' UTR begins at the transcription initiation site and continues up to, but not including, the start codon, while the 3' UTR begins immediately after the stop codon and continues until the transcription termination signal. Various lines of evidence have demonstrated that UTRs play a role in regulating the stability and translation of nucleic acid molecules. Regulatory properties of UTRs may be incorporated into the polynucleotides of the present disclosure, particularly to enhance molecular stability. Specific features can also be incorporated to ensure controlled downregulation of transcripts if they are misdirected to undesirable organ sites. Multiple 5' UTR and 3' UTR sequences are known and available in the art.

[0101] In some embodiments of the present disclosure, the 5' UTR is a heterologous UTR, i.e., a UTR found in nature associated with a different ORF. In another embodiment, the 5' UTR is a synthetic UTR, i.e., not found in nature. Synthetic UTRs include UTRs that have been mutated to improve properties, such as UTRs that increase gene expression, and completely synthetic UTRs. Exemplary 5' UTRs include those derived from Xenopus or human alpha- or beta-globulin (US Pat. Nos. 8278063, 9012219), human cytochrome b-245a polypeptide, and hydroxysteroid (17b) dehydrogenase and tobacco etch virus (US Pat. Nos. 8278063, 9012219).

[0102] In some embodiments, a 5'UTR of the present disclosure comprises a sequence selected from SEQ ID NOs: 173-174.

[0103] In some preferred embodiments, a 5'UTR of the present disclosure comprises a sequence selected from SEQ ID NO:174.

[0104] The 3'UTR refers to the region immediately downstream (3') of the mRNA stop codon (the codon that signals the end of translation of the mRNA transcript). The 3'UTR does not encode a protein (it is non-coding). Native or wild-type 3'UTRs are known to contain embedded adenosine (A) and uridine (U) segments. These AU-rich imprints are particularly common in genes with high turnover rates.

[0105] The introduction, removal, or modification of 3'UTR AU-rich elements (AREs) can be used to modulate the stability of the nucleic acids (e.g., RNA) of the present disclosure. When engineering a particular nucleic acid, one or more copies of an ARE can be introduced to make the nucleic acid of the present disclosure less stable, thereby reducing translation and the resulting production of the protein. Similarly, AREs can be identified and removed or mutated to increase intracellular stability and thereby increase translation and production of the resulting protein.

[0106] The 3'UTR may be heterologous or synthetic. Regarding 3'UTRs, globulin UTRs (including Xenopus β-globulin UTRs and human β-globulin UTRs) are known in the art (8278063, 9012219, US20110086907). By cloning two consecutive human β-globulin 3'UTRs end-to-end, nucleic acids (e.g., mRNAs) encoding modified β-globulins with improved stability in several cell types have been developed, which are well known in the art (US2012 / 0195936, WO2014 / 071963).

[0107] In some embodiments, a 3'UTR of the present disclosure comprises a sequence selected from SEQ ID NO:175.

[0108] Those skilled in the art will appreciate that heterologous or synthetic 5'UTRs can be used with any desired 3'UTR sequence, for example, heterologous 5'UTRs can be used with synthetic 3'UTRs or heterologous 3'UTRs.

[0109] It should be understood that any UTR from any gene can be incorporated into a region of a nucleic acid. Furthermore, multiple wild-type UTRs from any known gene can be utilized. It is also within the scope of the present disclosure to provide artificial UTRs that are not wild-type regions. These UTRs, or portions thereof, can be positioned in the same orientation within the selected transcript, or their orientation or position can be changed. Thus, a 5' or 3' UTR can be inverted, shortened, extended, or prepared with one or more other 5' or 3' UTRs. When referring to a UTR sequence, the term "modified" as used herein means that the UTR has been altered in some way relative to the reference sequence. For example, a 3' or 5' UTR can be altered compared to a wild-type or native UTR by a change in orientation or position, as taught above, or by the incorporation of additional nucleotides, deletion of nucleotides, nucleotide exchange, or rearrangement. Any one of these alterations that produces an "altered" UTR (either 3' or 5') includes a mutant UTR.

[0110] In vitro transcription of RNA: cDNA encoding the polynucleotides described herein can be transcribed using an in vitro transcription (IVT) system. In some embodiments, the RNA transcripts are produced in an in vitro transcription reaction using an unamplified, linearized DNA template to produce the RNA transcript. In some embodiments, the template DNA is isolated DNA. In some embodiments, the template DNA is cDNA. In some embodiments, the cDNA is formed by enzymatic digestion of circular plasmid DNA. In some embodiments, a plasmid DNA template is used to transfect cells, such as bacterial cells (e.g., E. coli, such as DH-1 cells). In some embodiments, the transfected cells are cultured to replicate the plasmid DNA, which is then isolated and purified. In some embodiments, the DNA template comprises an RNA polymerase promoter, such as a T7 promoter, located 5' to and operably linked to a target gene.

[0111] In some embodiments, the in vitro transcription template encodes a 5' untranslated (UTR) region, contains an open reading frame, encodes a 3' UTR, and a poly(A) tail. The specific nucleic acid sequence composition and length of the in vitro transcription template depend on the mRNA encoded by the template.

[0112] The "5' untranslated region" (UTR) is the region immediately upstream (i.e., 5') of the start codon of an mRNA (i.e., the first codon of an mRNA transcript that is translated by the ribosome) that does not encode a polypeptide. When producing an RNA transcript, the 5' UTR may contain a promoter sequence. Such promoter sequences are known in the art. It is understood that such promoter sequences are not present in the vaccines of the present disclosure.

[0113] "3' untranslated region" (UTR) refers to the region immediately downstream (ie, 3') of the stop codon of an mRNA (ie, the codon that signals the termination of translation of the mRNA transcript), that does not encode a polypeptide.

[0114] An "open reading frame" is a contiguous DNA segment beginning with a start codon (e.g., methionine (ATG)) and ending with a stop codon (e.g., TAA, TAG, or TGA) that encodes a polypeptide.

[0115] A "poly(A) tail" is a region of an mRNA located downstream, such as immediately downstream (i.e., 3') of the 3' UTR, that contains multiple consecutive adenosine monophosphates. The poly(A) tail may contain 10 to 300 adenosine monophosphates. For example, the poly(A) tail may contain 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, or 300 adenosine monophosphates. In some embodiments, the poly(A) tail contains 50 to 250 adenosine monophosphates. In relevant biological environments (e.g., intracellular, in vivo), the poly(A) tail serves to protect mRNA from enzymatic degradation in the cytoplasm, facilitate the termination of transcription, and / or aid in the transport and translation of mRNA from the nucleus.

[0116] In some embodiments, the nucleic acid comprises 200 to 3,000 nucleotides. For example, the nucleic acid may comprise 200 to 500, 200 to 1,000, 200 to 1,500, 200 to 3,000, 500 to 1,000, 500 to 1,500, 500 to 2,000, 500 to 3,000, 1,000 to 1,500, 1,000 to 2,000, 1,000 to 3,000, 1,500 to 3,000, or 2,000 to 3,000 nucleotides.

[0117] In vitro transcription systems typically include a transcription buffer, nucleotide triphosphates (NTPs), an RNase inhibitor, inorganic pyrophosphate, and a polymerase.

[0118] The NTPs may be prepared in-house, selected from a supplier, or synthesized as described herein, including, but not limited to, the NTPs described herein, including natural and non-natural (modified) NTPs.

[0119] Any number of RNA polymerases or mutants can be used in the methods of the present disclosure. The polymerase may be selected from bacteriophage RNA polymerases, such as, but not limited to, T7 RNA polymerase, T3 RNA polymerase, SP6 RNA polymerase, and / or mutant polymerases, including, but not limited to, polymerases capable of incorporating modified nucleic acids and / or modified nucleotides (including chemically modified nucleic acids and / or nucleotides). In some embodiments, the use of DNase is excluded.

[0120] In some embodiments, the RNA transcripts can be capped by enzymatic capping or co-transcriptional capping reactions. In some embodiments, the RNA comprises a 5'-end cap, e.g., 7mG(5')ppp(5')NlmpNp.

[0121] Purification of mRNA: The purification of nucleic acids described herein includes, but is not limited to, nucleic acid purification, quality assurance, and quality control. Purification can be performed by methods known in the art, including, but not limited to, lithium chloride precipitation, beads (Beckman Coulter Genomics, Danvers, MA), poly-T beads, LNA™ oligo-T capture probes (Vedbaek, Denmark), or HPLC-based purification methods such as strong anion exchange HPLC, weak anion exchange HPLC, reverse-phase HPLC (RP-HPLC), and hydrophobic interaction HPLC (HIC-HPLC). When used with respect to nucleic acids, the term "purified," such as "purified nucleic acid," refers to a nucleic acid that has been separated from at least one contaminant. A "contaminant" is any substance that renders another substance unsuitable, impure, or inferior. Thus, purified nucleic acids (e.g., DNA and RNA) exist in a form or environment different from that found in nature or that existed prior to the processing or purification method.

[0122] Quality assurance and / or quality control checks can be performed using methods such as, but not limited to, gel electrophoresis, UV absorbance, or analytical HPLC.

[0123] Quantification: In some embodiments, nucleic acids can be quantified using methods such as, but not limited to, ultraviolet-visible spectroscopy (UV / Vis). A non-limiting example of a UV / Vis spectrometer is a spectrometer (ThermoFisher, Waltham, MA). Quantified nucleic acids can be analyzed to determine whether the nucleic acids are of the appropriate size to confirm that the nucleic acids have not been degraded. Nucleic acid degradation can be confirmed by methods including, but not limited to, HPLC-based purification methods, including, but not limited to, agarose gel electrophoresis, strong anion exchange HPLC, weak anion exchange HPLC, reverse-phase HPLC (RP-HPLC), and hydrophobic interaction HPLC (HIC-HPLC), liquid chromatography-mass spectrometry (LCMS), capillary electrophoresis (CE), and capillary gel electrophoresis (CGE).

[0124] Lipid Nanoparticles (LNPs): In some embodiments, the RNA (e.g., mRNA) of the present disclosure is formulated into lipid nanoparticles (LNPs). Lipid nanoparticles typically include an ionizable cationic lipid, a non-cationic lipid, a sterol and PEG lipid fraction, and a target nucleic acid material. The lipid nanoparticles of the present disclosure can be produced by ingredients, compositions, and methods commonly known in the art, see, e.g., CN116535381B, TW202340136A, CN116178193B, CN116178193B, CN115784921B, CN115745820B, CN115677518B, CN114957027B, CN114044741B, CN116854754A, CN116785265A, CN116789764A, CN116396178A, CN116375592A, CN116082275A, each of which is incorporated herein by reference in its entirety.

[0125] As used herein, the term "cationic lipid" refers to the lipid that is positively charged at selected pH value.Cationic lipid can easily bind to negatively charged nucleic acid, that is, form lipid nanoparticle (LNP) through electrostatic interaction with the negatively charged phosphate group present in nucleic acid.LNP is currently one of the most popular transport carriers.

[0126] As used herein, the term "neutral lipid" refers to an auxiliary lipid that exists in an uncharged or zwitterionic form at a selected pH value, which may modulate the fluidity of nanoparticles into a lipid bilayer structure by promoting lipid phase transition, improving efficacy and potentially affecting target organ specificity.

[0127] As used herein, "structured lipid" refers to a lipid that enhances the stability of nanoparticles by filling the interstices between lipids. The structured lipid may be selected from the group consisting of, but not limited to, cholesterol, nonsterols, sitosterol, ergosterol, campesterol, stigmasterol, brassisterol, tomatine, ursolic acid, α-tocopherol, corticosteroids, and mixtures thereof. In some embodiments, the structured lipid is cholesterol. In some embodiments, the structured lipid comprises cholesterol and a corticosteroid (e.g., prednisolone, dexamethasone, prednisone, and hydrocortisone), or any combination thereof.

[0128] In some embodiments, the molar ratio of the cationic lipid to the structural lipid is about 1:1 to 5:1, e.g., about 1.0:1, 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2.0:1.

[0129] In some embodiments, the molar ratio of the cationic lipid to the neutral lipid is about (1-10):1, for example, about 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1.

[0130] In some embodiments, the molar ratio of the cationic lipid to the structural lipid is about (1-5):1, for example, about 1:1, 2:1, 3:1, 4:1, or 5:1.

[0131] In some embodiments, the molar ratio of the cationic lipid, the neutral lipid, the structured lipid, and the polymer-conjugated lipid is (25-65):(5-25):(25-45):(0.5-5). For example, in some embodiments, the ratio (relative molar ratio) of the cationic lipid is 25 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts, 55 parts, 60 parts, or 65 parts, the neutral lipid is 5 parts, 10 parts, 15 parts, 20 parts, or 25 parts, the structured lipid is 25 parts, 30 parts, 35 parts, 40 parts, or 45 parts, and the polymer-conjugated lipid is 0.5 parts, 1 part, 1.5 parts, 2 parts, 2.5 parts, 3.0 parts, 3.5 parts, 4.0 parts, 4.5 parts, or 5.0 parts.

[0132] In some embodiments, the molar ratio of the cationic lipid, the neutral lipid, the structured lipid, and the polymer-conjugated lipid is (49-50):10:(38.5-39.5):1. For example, in some embodiments, the ratio (relative molar number) is 49 or 50 parts cationic lipid, 10 parts neutral lipid, 38.5 parts, 39.0 parts, or 39.5 parts structured lipid, and 1 part polymer-conjugated lipid, e.g., 50:10:38.5:1.5 or 49:10:39.5:1.5.

[0133] As used herein, the term "polymer-conjugated lipid" refers to a lipid modified with polyethylene glycol (PEG). Hydrophilic PEG stabilizes LNPs, regulates nanoparticle size by limiting lipid fusion, and extends the half-life of nanoparticles by reducing nonspecific interactions with macrophages. In some embodiments, the polymer-conjugated lipid is selected from one or more of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, and PEG-modified dialkylglycerol. The molecular weight of the PEG modification is typically 350 to 5000 Da. For example, the polymer-conjugated lipid is distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE). PEG2000), 1,2-dimyristoyl-rac-glycero-methoxypolyethylene glycol-2000 (DMG PEG2000), and methoxypoly(ethylene glycol) ditetradecylacetamide (methoxypoly(ethylene glycol) ycol)ditetradecylacetamide) (ALC0159).

[0134] In one embodiment of the composition / carrier of the present disclosure, the polymer-conjugated lipid is DMG-PEG2000.

[0135] In one embodiment of the composition / carrier of the present disclosure, the carrier comprises a cationic lipid, a neutral lipid, a structured lipid, and a polymer-conjugated lipid, and the molar ratio of the cationic lipid, the neutral lipid, the structured lipid material, and the polymer-conjugated lipid is (25-65):(5-25):(25-45):(60.5-5), for example, (45-55):(9-11):(34-43):(0.5-2.5).

[0136] In one embodiment of the composition / carrier of the present disclosure, the carrier comprises a cationic lipid, a neutral lipid, a structured lipid, and a polymer-conjugated lipid, wherein the molar ratio of the cationic lipid, the neutral lipid, the structured lipid material to the polymer-conjugated lipid is 50:10:38.5:1.5.

[0137] As used herein, the term "alkyl" is intended to include both branched and straight-chain saturated aliphatic monovalent hydrocarbon groups having the specified number of carbon atoms. As used in this disclosure, the term "alkylene" is intended to include both branched and straight-chain saturated aliphatic divalent hydrocarbon groups having the specified number of carbon atoms. n~m indicates a group having n to m carbon atoms. For example, C 2~5 Alkylene includes C2 alkylene, C3 alkylene, C4 alkylene, and C5 alkylene. 2~8 Alkylene includes C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, C6 alkylene, C7 alkylene, and C8 alkylene. 1~6Alkylene includes C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, and C6 alkylene. 1~3 Alkylene includes C1 alkylene, C2 alkylene, and C3 alkylene. 6~15 Straight chain alkyls include those having 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 carbon atoms. 12~25 Branched alkyls include branched alkyls having 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 carbon atoms. An alkyl (or alkylene) can be unsubstituted or it can be substituted, where at least one hydrogen is replaced with another chemical group.

[0138] As used herein, a "therapeutically effective amount" refers to the amount of a therapeutic agent that can ameliorate a disease or symptom when administered to a patient. A "prophylactically effective amount" refers to the amount of a prophylactic agent that can prevent a disease or symptom when administered to a subject. The amount of a therapeutic agent that constitutes a "therapeutically effective amount" or the amount of a prophylactic agent that constitutes a "prophylactically effective amount" varies depending on the therapeutic / prophylactic agent, the state and severity of the disease, and the age, weight, etc. of the patient / subject to be treated / prevented. Therapeutically effective amounts and prophylactically effective amounts can be routinely determined by one of ordinary skill in the art based on their knowledge and this disclosure.

[0139] The compositions herein can be used to induce a protective immune response against VZV in a subject in need thereof, the protective immune response including, for example, the production of neutralizing antibodies. In some embodiments, the subject is immunocompromised. The subject is 10 years of age or older, for example, 45, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80 years of age.

[0140] The vaccines of the present disclosure are typically formulated in lipid nanoparticles. In some embodiments, the lipid nanoparticles contain at least one ionizable cationic lipid, at least Each of the lipids contains one non-cationic lipid, at least one sterol, and / or at least one polyethylene glycol (PEG)-modified lipid.

[0141] In some preferred embodiments, the lipid nanoparticles comprise cationic lipids, neutral lipids, structured lipids, and polymer-conjugated lipids.

[0142] In some preferred embodiments, the cationic lipid is a compound of the structure of Formula I, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein G is C 1~6 alkylene, and G2 is C 2~8 alkylene, and G3 is C 1~3 alkylene, and L1 is C 6~15 is a straight chain alkyl, and L2 is C 12~25 It is a branched alkyl, for example, YK-009 (see Patent CN114044741B) of the formula II structure.

[0143] [ka]

[0144] In some preferred embodiments, the cationic lipid is a compound of the structure of Formula II, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein G is C 2~8 alkylene, and G2 is C 2~8 alkylene, L1 is -C(O)O- or -OC(O)-, L2 is -C(O)O- or -OC(O)-, and R1 is C 6~25 is a straight or branched chain alkyl, and R2 is C 6~25It is a straight-chain or branched alkyl, G3 is HO(CH2)2- or HO(CH2)3-, G4 is HO(CH2)2- or HO(CH2)3-, and L is (CH2)2-, -(CH2)3-, or -(CH2)4-. For example, YK-401 having the structure of formula II-I and YK-402 having the structure of formula II-II (see Patent CN115784921B).

[0145] [ka]

[0146] In some preferred embodiments, the cationic lipid is a compound of the structure of Formula III, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein G is C 2~8 alkylene, and G2 is C 2~8 alkylene, and R1 is C 6~20 is a straight or branched chain alkyl, and R2 is C 12~25 G3 is a linear alkyl, and G3 is HO(CH2)2N(CH3)(CH2)2-, HO(CH2)2N(CH2CH3)(CH2)2)-, (HO(CH2)2)2N(CH2)2)-, CHO(CH2)2N(CH3)(CH2)2), (CH3)2N(CH2)3SC(O)O(CH2)2)-, (CH3)2N(CH2)3SC(O))-, CH3NH(CH2)2N(CH3)(CH2)2) or CH3CH2NH(CH2)2-. For example, YK-201 having the structure of formula III-I and YK-202 having the structure of formula III-II (see Patent CN115677518B).

[0147] [ka]

[0148] In some preferred embodiments, the cationic lipid is a compound of the structure of Formula IV, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein G is C 1~8 alkylene, and G2 is C 2~8alkylene, and R1 is C 6~20 is a straight or branched chain alkyl, and R2 is C 12~25 G is a straight-chain or branched alkyl, and G is HO(CH)N(R)CHCH(OH)CH-, where R is -CH, -CHCH, or -CHCHOH. For example, YK-305 has the structure of formula IV-I, and YK-310 has the structure of formula IV-II (see patent CN115745820B).

[0149] [ka]

[0150] In some preferred embodiments, the cationic lipid is a compound of the structure of Formula V, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein G 1 and G 2 are each independently unsubstituted C6 to C 10 alkylene, and G 3 is unsubstituted C1~C 12 alkylene, and R 1 and R 2 are C6 to C 24 Alkyl or C6-C 24 alkenyl, and R 3 is OR 5 , N, -C(=O)OR 4 , -OC(=O)R 4 or -NR 5 C(=O)R 4 and R 4 is C1~C 12 is hydrocarbyl, and R 5 is H or C1-C6 hydrocarbyl, such as ALC0315 (see patent CN108368028B) with the structure of formula VI.

[0151] [ka]

[0152] In some preferred embodiments, the cationic lipid is a compound of the structure of Formula VI, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein R is —(—(CH) n Q and -(CH2) n CHQR, where Q is -OR, -OH, or -O(CH2) n It is selected from the group consisting of N(R)2, -OC(O)R, -CX3, -CN, -N(R)C(O)R, -N(H)C(O)R, -N(R)S(O)R, -N(H)S(O)R, -N(R)C(O)N(R)2, -N(H)C(O)N(R)2, -N(H)C(O)N(H)(R), -N(R)C(S)N(R)2, -N(H)C(S)N(R)2, -N(R)S(O)R8 and heterocycle, wherein n is 1, 2 or 3, for example, SM102 of formula VI-I structure (see patent CN110520409A).

[0153] [ka]

[0154] In some preferred embodiments, the cationic lipid is a compound of the structure of Formula VII, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof.

[0155] [ka]

[0156] In some more preferred embodiments, the cationic lipids include YK-009, YK-401, YK-305, ALC0315, SM102, DLIN-MC3-DMA.

[0157] In some preferred embodiments, the molar ratio of the cationic lipid to the neutral lipid is 1:(1-10):1.

[0158] In some preferred embodiments, the molar ratio of the cationic lipid to the structural lipid is (1-5):1.

[0159] In some preferred embodiments, the molar ratio of the cationic lipid, the neutral lipid, the structured lipid, and the polymer-conjugated lipid is (25-75):(5-25):(15-65):(0.5-10).

[0160] In some preferred embodiments, the cationic lipid, the neutral lipid, the structure The molar ratio of the lipid to the polymer-conjugated lipid is (35-49):(7.5-15):(35-55):(1-5), for example, 49:10:43.5:1.5.

[0161] In some preferred embodiments, the neutral lipid comprises one or more of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, ceramide, sterol, and derivatives thereof.

[0162] In some preferred embodiments, the neutral lipid is 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-3-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16).0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoylphosphatidylglycerol (DPPG), palmitoyloleoylphosphatidylethanolamine (POPE), distearo ... In some embodiments, the phosphatidylcholine may be selected from one or more of sphingomyelin, phosphatidylethanolamine (DSPE), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), 1-stearoyl-2-oleoyl-stearoylethanolamine (SOPE), 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine (LPE), and mixtures thereof.

[0163] In some more preferred embodiments, the neutral lipid is DOPE and / or DSPC.

[0164] In some more preferred embodiments, the structured lipid is selected from one or more of cholesterol, a non-sterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid, α-tocopherol, and corticosteroids.

[0165] In some more preferred embodiments, the structured lipid is cholesterol.

[0166] In some more preferred embodiments, the polymer-conjugated lipid is a PEG-modified phosphatidylethanolamine, a PEG-modified phosphatidic acid, a PEG-modified ceramide, a PE The PEG-modified dialkylamine is selected from one or more of: PEG-modified diacylglycerol, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol.

[0167] In some more preferred embodiments, the polymer-conjugated lipid is selected from one or more of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), 1,2-dimyristoyl-rac-glycero-methoxypolyethylene glycol-2000 (DMG-PEG2000), and methoxypoly(ethylene glycol) ditetradecylacetamide (ALC-0159).

[0168] Pharmaceutical Preparations: The present specification provides compositions (e.g., pharmaceutical compositions), methods, kits, and reagents for preventing or treating varicella-zoster virus, e.g., in humans and other mammals. The compositions provided herein can be used as therapeutic or prophylactic agents. They can be used in medicines for the prevention and / or treatment of shingles.

[0169] In some embodiments, a varicella zoster vaccine comprising an RNA described herein can be administered to a subject (e.g., a mammalian subject, e.g., a human subject), and the RNA polynucleotide is translated in vivo to produce an antigenic polypeptide (antigen).

[0170] An "effective amount" of a composition (e.g., comprising RNA) is based, at least in part, on the target tissue, target cell type, method of administration, physical characteristics of the RNA (e.g., length, nucleotide composition, and / or degree of nucleoside modification), other components of the vaccine, and other determinants, such as the subject's age, weight, height, sex, and general health. Typically, an effective amount of a composition provides for the induction or enhancement of an immune response, which is modulated by antigen production in the subject's cells. In some embodiments, an effective amount of a composition comprising an RNA polynucleotide with at least one chemical modification is more effective than a composition comprising a corresponding unmodified polynucleotide encoding the same antigen or peptide antigen. Increased antigen production can be evidenced by increased cell transfection (percentage of cells transfected with the RNA vaccine), increased protein translation and / or expression from the polynucleotide, decreased nucleic acid degradation (e.g., increased duration of protein translation from the modified polynucleotide), or an alteration in the antigen-specific immune response of the host cell.

[0171] The term "pharmaceutical composition" refers to a combination of an active agent with an inert or active carrier such that the composition is particularly suitable for in vivo or ex vivo diagnostic or therapeutic use. A "pharmaceutically acceptable carrier" does not cause undesirable physiological effects after or when administered to a subject. A carrier of a pharmaceutical composition must be "acceptable" in the sense of being compatible with the active ingredient and capable of stabilizing the active ingredient. One or more solubilizing agents can be used as pharmaceutically acceptable carriers for delivering the active agent.

[0172] In some embodiments, compositions according to the present disclosure (including polynucleotides and polypeptides encoded thereby) can be used to treat or prevent shingles. The compositions may be administered prophylactically or therapeutically to healthy individuals as part of an active immunization regimen, or may be pre-administered early in infection during the incubation period or post-symptomatic active infection period. In some embodiments, the amount of RNA provided to a cell, tissue, or subject may be an amount effective for immunoprophylaxis.

[0173] In some embodiments, the composition may be administered by intramuscular injection.

[0174] The composition may be administered according to the prevalence of infectious diseases or the extent or level of unmet medical need. and may be used in a variety of settings. As a non-limiting example, RNA vaccines may be used to treat and / or prevent a variety of infectious diseases. Such RNA vaccines have superior properties because they generate much greater antibody titers, generate better neutralizing immunity, generate more durable immune responses, and / or generate responses more rapidly than commercially available vaccines.

[0175] The present disclosure provides pharmaceutical compositions comprising RNA and / or complexes, optionally in combination with one or more pharmaceutically acceptable excipients. In addition to conventional excipients (e.g., any and all solvents, dispersion media, diluents or other liquid vehicles, dispersing or suspending aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, etc.), excipients also include, but are not limited to, lipids, liposomes, lipid nanoparticles, polymers, lipid complexes, core-shell nanoparticles, peptides, proteins, cells transfected with RNA (e.g., for implantation into a subject), hyaluronidase, nanoparticle mimics, and combinations thereof.

[0176] The RNA may be formulated or administered alone or in combination with one or more other components, for example, an immunological composition may include other components, including, but not limited to, an adjuvant.

[0177] The relative amounts of active ingredient, pharmaceutically acceptable excipient, and / or any additional ingredients of a pharmaceutical composition according to the present disclosure will further vary depending on the identity, size, and / or condition of the subject being treated, as well as the route by which the composition is administered. For example, the composition may comprise between 0.1% and 100%, e.g., between 0.5% and 50%, between 1% and 30%, between 5% and 80%, or at least 80% (w / w) active ingredient.

[0178] In some embodiments, the RNA is formulated with one or more excipients to impart functions such as increasing mRNA stability, increasing cell transfection efficiency and protein translation efficiency, or altering the biodistribution of the mRNA to target specific tissues or cell types.

[0179] Administration / Application: Provided herein are immunological compositions (e.g., RNA vaccines), methods, kits, and reagents for preventing varicella-zoster virus infection in humans and other mammals. The immunological compositions may be used as therapeutic or prophylactic agents. In some embodiments, the immunological compositions are used to provide prophylactic protection against shingles.

[0180] The subject may be any mammal, including non-human primates and human subjects.

[0181] In some embodiments, the immunological composition (e.g., mRNA vaccine) is administered to a subject (e.g., a mammalian subject, e.g., a human subject) in an amount effective to induce an antigen-specific immune response. The RNA encoding the VZV gE antigen is expressed and translated in vivo to produce the antigen, which then stimulates an immune response in the subject.

[0182] Prophylactic protection against shingles can be achieved after administration of the immunological compositions (e.g., mRNA vaccines) of the present disclosure. The immunological compositions may be administered one, two, three, four, or more times, although a single administration of the vaccine (optionally followed by a single booster) may be sufficient.

[0183] In accordance with aspects of the present disclosure, methods are provided for eliciting an immune response to a varicella-zoster virus antigen (or antigens) in a subject. In some embodiments, the methods include administering to the subject an immunogenic composition comprising RNA (e.g., mRNA) having an open reading frame encoding a VZV gE glycoprotein, thereby inducing varicella-zoster in the subject. The vaccine involves inducing an immune response specific to the herpes virus antigen response, and the subject's anti-antigen antibody titer after vaccination is increased compared to the anti-antigen antibody titer of a subject vaccinated with a conventional vaccine (e.g., GSK's recombinant subunit vaccine). An "anti-antigen antibody" is a serum antibody that specifically binds to an antigen.

[0184] A prophylactically effective amount is an amount effective to prevent viral infection at a clinically acceptable level. In some embodiments, an effective dose is the dosage described in the instructions or package insert for the vaccine. As used herein, a conventional vaccine refers to a vaccine other than the mRNA vaccine of the present disclosure. For example, conventional vaccines include, but are not limited to, live microbial vaccines, killed microbial vaccines, subunit vaccines, protein antigen vaccines, DNA vaccines, virus-like particle (VLP) vaccines, etc. In exemplary embodiments, a conventional vaccine is one that has received regulatory approval and / or is registered with a national drug regulatory agency (e.g., the U.S. Food and Drug Administration (FDA) or the European Medicines Agency (EMA)).

[0185] In another embodiment of the present disclosure, there is provided a method for inducing an immune response against varicella-zoster virus in a subject, the method comprising administering to the subject an immunological composition (e.g., an RNA vaccine) comprising an RNA polynucleotide comprising an open reading frame encoding a VZV gE antigen, thereby inducing an immune response specific to varicella-zoster virus in the subject.

[0186] In some other embodiments, the immune response is assessed by measuring proteins (antibody titers) in the subject. In other embodiments, serum or antibodies from immunized subjects are tested for their ability to neutralize viral uptake or attenuate varicella-zoster virus transformation of human B lymphocytes. In other embodiments, the ability to stimulate a strong T cell response is measured using techniques well known in the art.

[0187] The present specification also provides a method for inducing an immune response to varicella-zoster virus in a subject by administering to the subject RNA having an open reading frame encoding a first antigen, wherein the RNA does not contain a stabilizing element, and wherein an adjuvant is not co-formulated or co-administered with the vaccine.

[0188] The immunological composition (e.g., RNA vaccine) may be administered by any route that produces a therapeutically effective result. These routes include, but are not limited to, intradermal, intramuscular, or subcutaneous administration. The present disclosure provides methods comprising administering an RNA vaccine to a subject in need thereof. The exact amount required will vary from subject to subject, depending on the subject's species, age, and general condition, the severity of the disease, the particular composition, its method of administration, its mode of activity, and the like. RNA is typically formulated in unit dosage form for ease of administration and uniformity of dosage. However, it should be understood that the total daily dose of RNA may be determined by the attending physician within the scope of sound medical judgment. The specific therapeutically effective, prophylactically effective, or appropriate imaging dosage level for a particular patient will depend on a variety of factors, including the disease and severity of the disease being treated, the activity of the specific compound used, the specific composition used, the patient's age, weight, general health, sex, and diet, the time of administration, the route of administration, and the excretion rate of the specific compound used, the duration of treatment, the composition of the specific compound used or co-administered drugs, and similar factors well known in the medical field.

[0189] An effective amount of the RNA provided herein can be as low as 5 μg, e.g., administered in a single dose or two 2.5 μg doses. In some embodiments, an effective amount is a total dose of 5 μg to 200 μg. For example, effective amounts include 5 μg, 10 μg, 20 μg, 25 μg, 30 μg, 35 μg, 40 μg, 45 μg, 50 μg, 55 μg, 60 μg, 6 The total dose may be 5 μg, 70 μg, 75 μg, 80 μg, 85 μg, 90 μg, 95 μg, 100 μg, 110 μg, 120 μg, 130 μg, 140 μg, 150 μg, 160 μg, 170 μg, 180 μg, 190 μg, or 200 μg. In some embodiments, the effective amount is a total dose of 5 μg to 200 μg. In some embodiments, the effective amount is a total dose of 10 μg. In some embodiments, the effective amount is a total dose of 20 μg. In some embodiments, the effective amount is a total dose of 75 μg. In some embodiments, the effective amount is a total dose of 150 μg. In some embodiments, the effective amount is a total dose of 200 μg.

[0190] Vaccine effectiveness Some aspects of the present disclosure provide formulations of immune compositions (e.g., RNA vaccines), wherein the RNA is formulated in an amount effective to generate an antigen-specific immune response in a subject (e.g., to generate antibodies specific to a varicella-zoster virus antigen). An "effective amount" is the amount of RNA administered that is effective to generate an antigen-specific immune response. The present disclosure further provides methods of inducing an antigen-specific immune response in a subject.

[0191] As used herein, an immune response to a vaccine or LNP of the present disclosure is the generation of a humoral and / or cellular immune response in a subject to the varicella-zoster virus protein(s) present in the vaccine. For purposes of this disclosure, a "humoral" immune response refers to an immune response mediated by antibody molecules (e.g., including secretory (IgA) or IgG molecules), whereas a "cellular" immune response refers to an immune response mediated by T lymphocytes (e.g., CD4 + Helper cells and / or CD8 +Cellular immunity refers to immune responses mediated by T cells (e.g., CTLs) and / or other white blood cells. An important aspect of cellular immunity includes antigen-specific responses elicited by cytolytic T cells (CTLs). CTLs are specific for peptide antigens encoded by the major histocompatibility complex (MHC) and presented in association with proteins expressed on the cell surface. CTLs are instrumental in inducing and promoting the destruction of intracellular microorganisms or the lysis of cells infected with such microorganisms. Another aspect of cellular immunity includes antigen-specific responses elicited by helper T cells. Helper T cells are used to stimulate the function and focus the activity of nonspecific effector cells to cells that present peptide antigens on their surface in association with MHC molecules. Cellular immune responses elicit the production of cytokines (e.g., IFN-γ, IL-2, TNF-β), chemokines, and other such molecules produced by activated T cells and / or other white blood cells.

[0192] In some embodiments, the antigen-specific immune response is characterized by measuring the anti-varicella-zoster virus antigen antibody titer produced in a subject administered an immunological composition provided herein. The antibody titer is a measurement of the amount of antibodies (e.g., antibodies specific to a particular antigen (e.g., anti-VZV gE glycoprotein) or epitope of an antigen) in a subject. The antibody titer is usually expressed as the reciprocal of the highest dilution that provides a positive result. For example, an enzyme-linked immunosorbent assay (ELISA) is a common assay used to measure antibody titers.

[0193] In some embodiments, antibody titers are used to determine whether a subject is infected or in need of immunization. In some embodiments, antibody titers are used to determine the strength of an autoimmune response, determine the need for booster vaccinations, determine whether a previous vaccine was effective, and identify recent or previous infections. According to the present disclosure, antibody titers may be used to determine the strength of the immune response elicited in a subject by an immune composition (e.g., an RNA vaccine).

[0194] In some embodiments, the anti-VZV gE antigen antibody titer produced in the subject is The antibody titer produced in the subject increases by at least 1 log compared to a control (a subject that has not been vaccinated). For example, the anti-VZV gE antigen antibody titer produced in the subject may increase by at least 1.5 log, at least 2 log, at least 2.5 log, at least 3 log, at least 3.5 log, or at least 4 log compared to a control. In some embodiments, the anti-VZV gE antigen antibody titer produced in the subject increases by 1, 1.5, 2, 2.5, or 3 logs compared to a control. In some embodiments, the anti-VZV gE antigen antibody titer produced in the subject increases by 1 to 4 logs compared to a control. For example, the anti-VZV gE antigen antibody titer produced in the subject may increase by 1 to 1.5, 1 to 2, 1 to 2.5, 1 to 3, 1.5 to 2, 1.5 to 2.5, 1.5 to 3, 2 to 2.5, 2 to 3, or 2.5 to 4 logs compared to a control.

[0195] In some embodiments, the anti-VZV gE antigen antibody titer produced in the subject is increased by at least 2-fold compared to a control (vaccinated with the conventional vaccine GSK Shingrix®). For example, the anti-VZV gE antigen antibody titer produced in the subject may be increased by at least 1-fold, at least 1.5-fold, at least 2-fold, at least 2.5-fold, or at least 3-fold compared to the control GSK.

[0196] In some embodiments, the control is an anti-Varicella-Zoster virus antigen antibody titer produced in a subject who has not received an immunological composition (e.g., an RNA vaccine). In some embodiments, the control is an anti-Varicella-Zoster virus antigen antibody titer produced in a subject who has received the recombinant subunit vaccine Shingrix®.

[0197] In some embodiments, the efficacy of an immune composition (e.g., an RNA vaccine) is measured in a mouse model. For example, an immune composition can be administered to a mouse model and the induction of neutralizing antibody titers in the mouse model can be determined. Viral challenge studies can also be used to evaluate the efficacy of vaccines of the present disclosure. For example, an immune composition can be administered to a mouse model, the mouse model can be challenged with a virus, and the survival and / or immune response (e.g., T cell response (e.g., cytokine response)) of the mouse model can be measured.

[0198] The present invention further relates to the following embodiments.

[0199] 1. comprising ribonucleic acid (RNA) of varicella-zoster virus (VZV), said RNA encoding wild-type VZV gE glycoprotein or a mutant thereof; wherein the sequence of the wild-type VZV gE glycoprotein is set forth in SEQ ID NO:3.

[0200] 2. The composition of embodiment 1, wherein the VZV gE glycoprotein variant comprises one or more of the following mutations compared to the wild-type VZV gE glycoprotein: sequence truncation, site mutation, and sequence deletion.

[0201] 3. The composition of embodiment 1 or 2, wherein the sequence truncation comprises the absence of a carboxyl-terminal tail domain.

[0202] 4. The sequence truncation is selected from the group consisting of: 1) truncation of amino acids 540 to 623 from the C-terminus of the gE protein, i.e., retention of amino acids 1 to 539 of the VZV gE protein; 2) truncation of amino acids 569 to 623 from the C-terminus of the gE protein, i.e., retention of amino acids 1 to 568 of the VZV gE protein; 3) truncation of the amino acids 574 to 623 from the C-terminus of the gE protein, i.e., retention of the amino acids 1 to 573 of the VZV gE protein; 4) truncation of amino acids 588 to 623 from the C-terminus of the gE protein, i.e., retention of amino acids 1 to 587 of the VZV gE protein; 5) truncation of amino acids 602 to 623 from the C-terminus of the gE protein, i.e., retention of amino acids 1 to 601 of the VZV gE protein; The composition of any one of embodiments 1 to 3, wherein the composition is selected from any one of:

[0203] 5. The VZV gE glycoprotein mutant comprises a site mutation selected from a mutation in a motif associated with ER retention, endocytosis, and / or a mutation at least one site in at least one motif associated with Golgi or trans-Golgi network localization of the gE protein, and a mutation at least one site in a gE C-terminal phosphorylated acidic motif; Here, the motif related to ER retention and endocytosis of the gE protein is "Y 582 A 583 G 584 L 585 " motif (e.g., motif Y 582 A 583 G 584 L 585 When a mutation occurs in , it can prevent endocytosis of the gE protein and reduce the localization of the gE antigen to the trans-Golgi network, for example, the mutation site Y582A, YK-VZV-013 (SEQ ID NO: 43), The motif associated with the localization of gE protein to the Golgi apparatus or trans-Golgi network is "A 568 Y 569 R 570 V 571 " motif (e.g., "A 568 Y 569 R 570 V 571 " motif, and a specific mutant antigen sequence is, for example, YK-VZV-011 (coordinate number 35)), The C-terminal oxidized acidic motif of gE is "S 593 E 594 S595 T 596 D 597 T 598 ” motif (e.g., “S 593 E 595 T 596 D 597 T 598 " motif is "A 593 E 594 A 596 D 597 A 598 " motif, which, when mutated, can reduce the localization of gE antigen to the trans-Golgi network; a specific mutant antigen sequence is, for example, YK-VZV-010 (SEQ ID NO: 31)).

[0204] 6. The composition of any one of embodiments 2 to 5, wherein the VZV gE glycoprotein variant comprises a site mutation selected from A568D, Y569K, Y569A, R570E, V571K, Y582A, S593A, S595A, T596A and T598A, e.g., selected from A568D, Y569K, R570E and V571K, e.g., selected from A568D and Y569K.

[0205] 7. The VZV gE glycoprotein mutant is 1)A568D, Y569K, R570E, V571K, (YK-VZV-023, YK-VZV-011) 2)A568D, Y569A, R570E, V571K, (YK-VZV-024) 3)Y569K, R570E, V571K, (YK-VZV-025) 4)A568D, Y569K, R570E, V571K, Y582A, (YK-VZV-030, YK-VZV-013) 5)A568D, Y569K, R570E, V571K, Y582G, (YK-VZV-031, YK-VZV-038) 6)A568D, Y582A, (YK-VZV-032) 7)Y569K, Y582A, (YK-VZV-033, YK-VZV-009) 8)R570E, Y582A, (YK-VZV-034) 9)V571K, Y582A, (YK-VZV-035) 10)S593A, S595A, T596A, T598A, (YK-VZV-016, YK-VZV-044) 11)Y582A, S593A, S595A, T596A, T598A, (YK-VZV-037) 12)A568D, Y569K, R570E, V571K, Y582G, S593A, S595A, T596A, T598A, (YK-VZV-039) 13)A568D, Y569K, R570E, V571K, Y582A, S593A, S595A, T596A, T598A, (YK-VZV-014) 14)Y582G, S593A, S595A, T596A, T598A, (YK-VZV-040) 15)Y569K, Y582A, S593A, S595A, T596A, T598A, (YK-VZV-010) 16)A568D, Y569K, R570E, V571K, S593A, S595A, T596A, T598A, (YK-VZV-012) 17) Y582G, (YK-VZV-029) 18) A568D, (YK-VZV-018) 19) Y569K, (YK-VZV-020) 20) R570E, (YK-VZV-021) 21) V571K, and (YK-VZV-022) 22)Y582A, (YK-VZV-028, YK-VZV-036) 7. The composition of any one of embodiments 2 to 6, comprising a site mutation selected from:

[0206] 8. The VZV gE glycoprotein mutant is 1)A568D, Y569K, R570E, V571K, (YK-VZV-011) 2)A568D, Y569A, R570E, V571K, (YK-VZV-024) 4)A568D, Y569K, R570E, V571K, Y582A, (YK-VZV-030, YK-VZV-013) 5)A568D, Y569K, R570E, V571K, Y582G, YK-VZV-031, YK-VZV-038) 7)Y569K, Y582A, (YK-VZV-009) 13)A568D, Y569K, R570E, V571K, Y582A, S593A, S595A, T596A, T598A, (YK-VZV-014) 15)Y569K, Y582A, S593A, S595A, T596A, T598A, (YK-VZV-010) 16)A568D, Y569K, R570E, V571K, S593A, S595A, T596A, T598A, (YK-VZV-012) 18) A568D, (YK-VZV-018) 19) Y569K, (YK-VZV-020) 20) R570E, (YK-VZV-021) and 22)Y582A(YK-VZV-028) The composition of any one of embodiments 2 to 7, comprising a site mutation selected from:

[0207] 9. The VZV gE glycoprotein mutant is 1) A 568 , 2) Y 569 RVDKSPYNQS 579 , and 3) Y 569 RVDKSPYNQSMYYAGLPV 587 9. The composition of any one of embodiments 2 to 8, comprising a sequence deletion selected from:

[0208] 10. The composition of any one of embodiments 1 to 9, wherein the VZV gE glycoprotein variant comprises a combination of site mutations, sequence truncations, and sequence deletions selected from the following:

[0209] [Table 1] TIFF2025179041000010.tif106169

[0210] 11. The composition of any one of embodiments 1 to 10, wherein the VZV gE glycoprotein variant comprises a combination of site mutations, sequence truncations and / or sequence deletions selected from the following:

[0211] [Table 2]

[0212] 12. The VZV gE glycoprotein variant has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 111%, 115, or 116. 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the sequence of any one of the preceding claims.

[0213] 13. The composition of any one of embodiments 1 to 12, wherein the VZV gE glycoprotein variant comprises SEQ ID NO: 31, 35, 43, 63 or 71.

[0214] 14. The composition of any one of embodiments 1 to 13, wherein the VZV RNA has an open reading frame (ORF) encoding a VZV gE glycoprotein or a variant thereof, and the sequence of the open reading frame has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 26, 30, 34, 38, 42, 46, 62, 70, 74, 86, 102, 110, 114 or 142.

[0215] 15. The composition of embodiment 14, wherein the sequence of the open reading frame is set forth in SEQ ID NO: 30, 34, 42, 62 or 70.

[0216] 16. The composition of any one of embodiments 1 to 15, wherein the VZV RNA further comprises a 5' untranslated region (UTR).

[0217] 17. The composition of embodiment 16, wherein the 5'UTR has the sequence set forth in SEQ ID NO: 173, 174, 175, 176 or 177.

[0218] 18. The composition of embodiment 17, wherein the 5'UTR has the sequence set forth in SEQ ID NO: 174.

[0219] 19. The composition of any one of embodiments 1 to 18, wherein the VZV RNA further comprises a 3' untranslated region (UTR).

[0220] 20. The composition of embodiment 19, wherein the 3'UTR has the sequence set forth in SEQ ID NO: 178, 179, 180 or 181.

[0221] 21. The composition of embodiment 20, wherein the 3'UTR has the sequence set forth in SEQ ID NO: 178.

[0222] 22. The composition of any one of embodiments 1 to 21, wherein the VZV RNA further comprises a poly(A) tail.

[0223] 23. The composition of embodiment 22, wherein the poly(A) tail has a length of 50 to 150 nucleotides.

[0224] 24. The composition of any one of embodiments 1 to 23, wherein the VZV RNA further comprises a 5'-end cap.

[0225] 25. The composition of embodiment 24, wherein the 5' end cap is 7mG(5')ppp(5')NlmpNp.

[0226] 26. The composition of any one of embodiments 14 to 25, wherein the sequence of the open reading frame is codon-optimized.

[0227] 27. The composition of embodiment 26, wherein the sequence of the open reading frame comprises at least one base modification.

[0228] 28. The composition of embodiment 27, wherein the base modification is selected from one or more of pseudouridine, N1-methylpseudouridine, N1-ethylpseudouridine, 2-thiouridine, 4'-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methoxyuridine and 2'-O-methyluridine.

[0229] 29. The composition of embodiment 28, wherein the base modification comprises replacing uracil with pseudouridine and / or N1-methylpseudouridine.

[0230] 30. The composition according to embodiment 27, 28, or 29, wherein the base modification is 1 to 100% base modification, for example, 1% base modification, 2% base modification, 3% base modification, 4% base modification, 5% base modification, 6% base modification, 7% base modification, 8% base modification, 9% base modification, 10% base modification, 15% base modification, 20% base modification, 25% base modification, 30% base modification, 35% base modification, 40% base modification, 45% base modification, 50% base modification, 55% base modification, 60% base modification, 65% base modification, 70% base modification, 75% base modification, 80% base modification, 85% base modification, 90% base modification, 95% base modification, and 100% base modification, and a range having any of the above values ​​as an endpoint.

[0231] 31. The composition of any one of embodiments 1 to 30, wherein the VZV RNA sequence has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 28, 32, 36, 40, 44, 48, 64, 72, 76, 88, 104, 112, 116 or 144.

[0232] 32. The composition of embodiment 31, wherein the VZV RNA sequence has the sequence set forth in SEQ ID NO: 32, 36, 44, 64 or 72.

[0233] 33. The composition of any one of embodiments 1 to 32, wherein the VZV RNA is mRNA.

[0234] 34. The composition of any one of embodiments 1 to 33, wherein the RNA encoding the VZV gE glycoprotein comprises an RNA sequence corresponding to a DNA sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 25, 29, 33, 37, 41, 45, 61, 69, 73, 85, 101, 109, 113 or 141.

[0235] 35. The composition of embodiment 34, wherein the RNA encoding the VZV gE glycoprotein comprises an RNA sequence corresponding to the DNA sequence set forth in SEQ ID NO: 29, 33, 41, 61, or 69.

[0236] 36. Providing a template capable of being transcribed into VZV RNA; transcribing using said template under conditions suitable for transcription into said RNA. 36. A method for preparing the composition of any one of embodiments 1 to 35, comprising:

[0237] 37. The method of embodiment 36, further comprising a purification step selected from lithium chloride precipitation, affinity chromatography, solution exchange by ultrafiltration, and cellulose chromatography.

[0238] 38. The composition of any one of embodiments 1-35, which is a vaccine and further comprises a pharmaceutically acceptable carrier.

[0239] 39. The composition of embodiment 38, wherein the carrier comprises a lipid mixture, the lipid mixture being, for example, a lipid nanoparticle (LNP).

[0240] 40. The composition of embodiment 38 or 39, wherein the vaccine is an mRNA vaccine.

[0241] 41. The composition of embodiment 39 or 40, wherein the lipid mixture is a lipid nanoparticle (LNP), the lipid nanoparticle comprising, for example, a cationic lipid, a neutral lipid, a structured lipid, and a polymer-conjugated lipid.

[0242] 42. The cationic lipid is a compound of formula I, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein G1 is C 1~6 alkylene, and G2 is C 2~8 alkylene, and G3 is C 1~3alkylene, and L1 is C 6~15 is a straight chain alkyl, and L2 is C 12~25 The composition of embodiment 41, wherein the cationic lipid is a branched alkyl, e.g., YK-009, having the structure of Formula II.

[0243] [ka]

[0244] 43. The cationic lipid is a compound of formula II, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein G1 is C 2~8 Al Kiren, G2, C 2~8 alkylene, L1 is -C(O)O- or -OC(O)-, L2 is -C(O)O- or -OC(O)-, and R1 is C 6~25 is a straight or branched chain alkyl; R2 is C 6~25 The composition of embodiment 41, wherein G3 is a straight or branched chain alkyl, G3 is HO(CH2)2- or HO(CH2)3-, G4 is HO(CH2)2- or HO(CH2)3-, and L is (CH2)2- or -(CH2)3- or -(CH2)4-, for example, the cationic lipid is YK-401 having the structure of formula II-I or YK-402 having the structure of formula II-II.

[0245] [ka]

[0246] 44. The cationic lipid is a compound of formula III, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein G1 is C 1~6 alkylene, and G2 is C 2~8 alkylene, and R1 is C 6~20 is a straight or branched chain alkyl; R2 is C 12~25The composition of embodiment 41, wherein G3 is a branched alkyl and G3 is HO(CH2)2N(CH3)(CH2)2-, HO(CH2)2N(CH2CH3)(CH2)2-, (HO(CH2)2)2N(CH2)2-, CHO(CH2)2N(CH3)(CH2)2-, (CH3)2N(CH2)3SC(O)O(CH2)2-, (CH3)2N(CH2)3SC(O)-, CH3NH(CH2)2N(CH3)(CH2)2- or CH3CH2NH(CH2)2-, for example, YK-201 having the structure of formula III-I or YK-202 having the structure of formula III-II.

[0247] [ka]

[0248] 45. The cationic lipid is a compound of formula IV, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein G1 is C 1~8 alkylene, and G2 is C 2~8 alkylene, and R1 is C 6~25 is a straight or branched chain alkyl; R2 is C 12~25 The composition of embodiment 41, wherein G3 is a straight or branched alkyl, and G3 is HO(CH2)2N(R3)CH2CH(OH)CH2-, where R3 is -CH3 or -CH2CH3 or -CH2CH2OH, such as YK-305 having the structure of formula IV-I, and YK-310 having the structure of formula IV-II.

[0249] [ka]

[0250] 46. ​​The cationic lipid is a compound of the structure of formula V, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein G 1 and G 2 are each independently unsubstituted C6 to C 10 alkylene, and G 3 is unsubstituted C1-C 12alkylene, and R 1 and R 2 are C6 to C 24 Alkyl or C6-C 24 alkenyl, and R 3 is OR 5 , N, -C(=O)OR 4 , -OC(=O)R 4 or -NR 5 C(=O)R 4 and R 4 is C1~C 12 is hydrocarbyl, and R 5 is H or C1-C6 hydrocarbyl, for example, ALC0315, structure of Formula VI.

[0251] [ka]

[0252] 47. The cationic lipid is a compound of formula VI, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof, wherein R4 is -(CH2) n Q and -(CH2) n CHQR, where Q is -OR, -OH, -O(CH2) n 42. The composition of embodiment 41, wherein n is selected from the group consisting of N(R), -OC(O)R, -CX, -CN, -N(R)C(O)R, -N(H)C(O)R, -N(R)S(O)R, -N(H)S(O)R, -N(R)C(O)N(R), -N(H)C(O)N(R), -N(H)C(O)N(H)(R), -N(R)C(S)N(R), -N(H)C(S)N(R), -N(R)S(O)R and heterocycle, wherein n is 1, 2 or 3, for example, SM102 of the structure of formula VI-I.

[0253] [ka]

[0254] 48. The composition of embodiment 41, wherein the cationic lipid is a compound of formula VII, or an N-oxide, solvate, pharmaceutically acceptable salt, or stereoisomer thereof.

[0255] [ka]

[0256] 49. The composition of embodiment 41, wherein the cationic lipid is selected from YK-009, YK-401, YK-305, ALC0315, SM102, and DLIN-MC3-DMA.

[0257] 50. The composition according to any one of embodiments 41 to 49, wherein the molar ratio of the cationic lipid to the neutral lipid is (1-10):1.

[0258] 51. The composition of any one of embodiments 41-50, wherein the molar ratio of the cationic lipid to the structural lipid is (1-5):1.

[0259] 52. The composition according to any one of embodiments 41 to 51, wherein the molar ratio of the cationic lipid, the neutral lipid, the structural lipid and the polymer-conjugated lipid is (25-65):(5-25):(25-45):(0.5-5).

[0260] 53. The molar ratio of the cationic lipid, the neutral lipid, the structural lipid, and the polymer-conjugated lipid is (49-50):10:(38.5-39.5):1.5, for example, 5 53. The composition of embodiment 52, wherein the ratio is 0:10:38.5:1.5 or 49:10:39.5:1.5.

[0261] 54. The composition of any one of embodiments 41 to 53, wherein the neutral lipid is selected from phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, ceramide, sterol, derivatives thereof, and any combination thereof.

[0262] 55. The neutral lipid may be 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distaloyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-3-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16).0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoylphosphatidylglycerol (DPPG), palmitoyloleoylphosphatidylethanolamine (POPE), distearoylphosphatidylethanolamine 55. The composition of any one of embodiments 41 to 54, wherein the phosphatidylcholine is selected from sphingomyelin, dipalmitoylphosphatidylethanolamine (DSPE), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), 1-stearoyl-2-oleoyl-phosphatidylethanolamine (SOPE), 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine (LPE), and mixtures thereof.

[0263] 56. The composition of any one of embodiments 41-55, wherein the neutral lipid is DOPE and / or DSPC.

[0264] 57. The composition of any one of embodiments 41-56, wherein the structured lipid is selected from sterols, cholesterol, non-sterols, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid, α-tocopherol, corticosteroids, and any combination thereof.

[0265] 58. The composition of embodiment 57, wherein the structured lipid comprises cholesterol, for example, is cholesterol.

[0266] 59. The polymer-conjugated lipid is PEG-modified phosphatidylethanolamine, PEG The composition of any one of embodiments 41 to 56, wherein the PEG-modified ceramide is selected from modified phosphatidic acid, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and any combination thereof.

[0267] 60. The composition of embodiment 59, wherein the polymer-conjugated lipid is selected from distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), 1,2-dimyristoyl-rac-glycero-methoxypolyethylene glycol-2000 (DMG-PEG2000), and methoxypoly(ethylene glycol) ditetradecylacetamide (methoxypoly(ALC-0159).

[0268] 61. The composition according to any one of embodiments 38 to 60, wherein the effective amount of VZV RNA is 25 μg to 200 μg, preferably 50 μg to 100 μg.

[0269] 62. The composition of any one of embodiments 38 to 60, wherein the vaccine is in an injectable form, for example a liquid formulation or a lyophilized formulation.

[0270] 63. A method for preparing a composition according to any one of embodiments 38 to 60, comprising mixing the VZV RNA with the pharmaceutically acceptable carrier, for example, encapsulating at least a portion of the RNA inside lipid nanoparticles.

[0271] 64. The method of embodiment 63, further comprising a purification step to remove non-encapsulated components, such as dialysis and / or filtration.

[0272] 65. The method of embodiment 64, wherein the unencapsulated component is selected from unencapsulated RNA, a non-aqueous solvent, and bacteria.

[0273] 66. Use of a composition described in any one of embodiments 1 to 62 in the preparation of a medicament for inducing a protective immune response against VZV in a subject, wherein the protective immune response includes, for example, the production of neutralizing antibodies.

[0274] 66a. The composition of any one of embodiments 1 to 62, for use in inducing a protective immune response against VZV in a subject in need thereof, wherein the protective immune response comprises, for example, the production of neutralizing antibodies.

[0275] 66b. A method of inducing a protective immune response against VZV in a subject in need thereof, comprising administering to said subject a composition described in any one of embodiments 1-62, wherein said protective immune response comprises, for example, the production of neutralizing antibodies.

[0276] 67. The use, composition or method of embodiment 66, 66a or 66b, wherein the subject is immunocompromised.

[0277] 68. The use, composition, or method of 66, 66a, or 66b, wherein the subject is 10 years of age or older, e.g., 50, 60, 70, 80 years of age or older.

[0278] 69. The use, composition or method of any one of embodiments 66 to 68, wherein the protective immune response further comprises a cellular immune response.

[0279] 70. The use, composition or method according to any one of embodiments 66 to 69, wherein the induced protective immune response is used to prevent VZV infection.

[0280] 71. The use, composition or method according to any one of embodiments 66 to 69, wherein said induced protective immune response is used for the prevention of VZV-associated pain.

[0281] Example 1: Design of VZV gE mutant antigens As a highly glycosylated type I membrane protein, gE can be transported between the endoplasmic reticulum (ER), the trans-Golgi network (TGN), and the endoplasmic reticulum. The gE protein contains several important domains related to its main functions.

[0282] (1) TM transmembrane domain: consisting of amino acids 539 to 559.

[0283] (2)A 568 YRV 571 Motif: Mediates transport of gE between the endoplasmic reticulum (ER), trans-Golgi network (TGN) and the endoplasm.

[0284] (3) Y 582 AGL 585 Motif: Mediates transport of gE between the endoplasmic reticulum (ER), trans-Golgi network (TGN) and the endoplasm.

[0285] (4)S 593 ES 595 T 596 DT 598 Motif: An important glycosylation site, mediating the transport of gE between the endoplasmic reticulum (ER), the trans-Golgi network (TGN), and the endoplasm.

[0286] The intracellular carboxyl terminus of gE plays a crucial role in the intracellular transport of gE. Based on the key domains of the gE protein and their functions, we designed a series of mRNA antigens targeting mutations in key structural and functional domains of gE, as well as various truncation mutations (see Figure 1).

[0287] Using the wild-type sequence of the gE gene of VZV virus (see NCBI virus strain sequence number QXN54923.1) as a reference, a series of 43 different gE mutant mRNA sequences were designed.

[0288] Table 1 summarizes mRNAs encoding mutant gE antigens with different C-terminal sequences. YK-VZV-004, YK-VZV-006, YK-VZV-045, and YK-VZV-007 in the table correspond to the corresponding mutations or combinations of mutations in the disclosed patents (US11643441B1, CN114081943A, US20230233671A1, and CN108472309A), respectively.

[0289] [Table 3] TIFF2025179041000020.tif222170TIFF2025179041000021.tif222170TIFF2025179041000022.tif171170

[0290] Example 2: Preparation of VZV gE antigen mRNA vaccine Preparation of transcription template linearized plasmid All antigen gene fragments were synthesized by Genscript Biotech Corporation and cloned into pVAX vector (purchased from Thermo Fisher Scientific) to successfully construct circular plasmids.

[0291] For template preparation, the plasmid was linearized using BsaI restriction endonuclease (purchased from Shanghai Beyotime Biotechnology Co., Ltd.), and the enzyme digestion system included the supercoiled plasmid inserted into the target gene corresponding to Example 1, 10x digestion buffer, BsaI restriction endonuclease, and RNase-free ddH2O. The enzyme digestion reaction temperature was 37°C, and the enzyme digestion time was 3 hours. The enzyme digestion system is as shown in Table 2.

[0292] [Table 4]

[0293] After the enzyme digestion reaction was completed, the linearized product was recovered using a DNA product purification kit (purchased from Yisheng Biotechnology Co., Ltd.), and the product concentration was measured using an ultra-micro UV spectrophotometer (Denovix). The length and state of the linearized plasmid template were detected through agarose gel.

[0294] Preparation of mRNA stock solution The in vitro co-transcriptional capping reaction is a process for preparing mRNA polynucleotides by assembling modified or natural nucleotide triphosphates (NTPs) according to the base sequence of a template. Adding Clean Cap to this process allows for the production of mRNA containing the Cap1 structure in a single transcription reaction. The typical reaction process consists of the following system (all purchased from Novoprotein Scientific Inc.):

[0295] [Table 5]

[0296] After the transcription was completed, 1 μl of DNase I (2 U / μl) was added, mixed uniformly, and incubated at 37°C for 20 min to remove template DNA. The transcription product was then purified using lithium chloride precipitation.

[0297] 1) To 20 μl of the reaction mixture, 30 μl of RNase-free H 2 O and 7.5 M lithium chloride (final concentration of lithium chloride is 2.8 M) was added.

[0298] 2) After uniform mixing, the mixture was left at -20°C for 2 hours, centrifuged at 12,000 rpm for 15 minutes, and the supernatant was discarded. The RNA precipitate was washed with 500 μl of 70% ethanol and centrifuged at 12,000 rpm for 5 minutes. This step was repeated once, and the precipitate was collected.

[0299] 3) After drying, 100 μl of enzyme-free water was added to dissolve the mRNA precipitate, and the purified RNA solution was stored at -80°C. The concentration and integrity of the mRNA were measured.

[0300] Preparation of mRNA-LNP (mRNA vaccine) Accurately weigh 20 mg each of cationic lipid, DSPC, cholesterol, and DMG-PEG2000, dissolve them in absolute ethanol, and prepare a 10 mg / ml solution. A mixed solution of the carriers was prepared according to a molar ratio of 49:10:39.5:1.5, which was used as the organic phase for later use.

[0301] VZV mRNA was dissolved in 50 mM citrate buffer, pH 4.0, and diluted to a 0.15 mg / ml solution, which served as the aqueous phase for subsequent use. A 3 ml BD (trade name, BiDi in Chinese; the same applies below) syringe was used to extract 2 ml of organic phase, and a 10 ml BD syringe was used to extract 7 ml of aqueous phase. The left pump pumped the organic phase, while the right pump pumped the aqueous phase. The LNP intermediate solution was then prepared in the microfluidic device.

[0302] Preparation volume: 9.0 ml, organic phase flow rate: 5.0 ml / min, aqueous phase flow rate: 15 ml / min, LNP intermediate solution was collected.

[0303] The LNP intermediate solution was diluted by adding 9 times the volume of PBS solution and concentrated by ultrafiltration using a small ultrafiltration device. The pore size of the ultrafiltration membrane was 50 KD, and when the solution was concentrated to approximately 10 ml, it was diluted by adding 4 times the volume of PBS solution and concentrated on the ultrafiltration device. Ultrafiltration was stopped when the final concentrated volume was less than 5 ml.

[0304] Example 3: Detection of varicella zoster mRNA vaccine antigen expression by Western Blot To verify whether the designed antigen mRNA sequences could be expressed in cells and the expression efficiency of various antigen sequences, cells were first transfected with the mRNA sequences and the expression of antigen mRNA in the cells was detected using Western blotting.

[0305] Experimental materials

[0306] [Table 6]

[0307] Experimental steps Cell culture and transfection: 2.5 x 10 293T cells were cultured in a 12-well plate. 5 The volume of mRNA preparation required for 500 ng of mRNA was calculated, and the corresponding volume of mRNA preparation was added directly to the cells and mixed evenly.

[0308] Experimental group: a mixture of protein samples obtained by transfecting cells with the series of mRNA vaccines prepared in Example 2 and then dissolving them; Negative control NC: RIPA lysate; Positive control PC: gE purified protein; Internal reference protein loading control: glyceraldehyde-3-phosphate dehydrogenase GAPDH, Protein sample preparation: (1) 16 hours after transfection, the cells were removed from the incubator, and all cells on the bottom of the dish were removed by repeated pipetting. The cells were then transferred to a 1.5 ml centrifuge tube and centrifuged at 1,500 rpm at 25°C for 3 minutes. (2) The supernatant was discarded, 1 ml of PBS was added, the cells were resuspended by pipetting, and the tubes were centrifuged at 1,500 rpm for 3 minutes. (3) Step (2) was repeated twice. (4) 50 μl of RIPA lysis solution (0.5 μl of 100× protein inhibitor was added to the RIPA lysis solution before use) was added to each cell tube, and the tubes were placed on ice for 30 minutes, during which time the samples were vortexed for 30 seconds every 5 minutes. (5) After pre-cooling the centrifuge and completely lysing the cells, the samples were centrifuged at 12,000 rpm at 4°C for 10 minutes. The supernatant was transferred to a new 1.5 ml centrifuge tube to obtain the test protein samples.

[0309] Development and analysis: Protein sample concentration was quantitatively determined using a BCA kit. 200 μl of BCA working solution and 10 μl of test sample were added. 10 μg of protein sample was pipetted into a 1.5 ml centrifuge tube, water was added to make a 24 μl system, 6 μl of 5x loading buffer was added, and the volume was adjusted to 30 μl with purified water. The total protein was denatured by boiling. 3 μg of sample (9 μl) was aspirated and spotted, and 4 μl of marker was spotted. Electrophoresis was performed at 200 V for 30 minutes. The film was transferred using a dry film transfer device, and the gel was removed to create a film transfer "sandwich." The negative plate was covered and voltages of 21V, 23V, and 25V were applied for 1 minute, 4 minutes, and 2 minutes, respectively, followed by blocking at room temperature for 1 hour. Primary antibody incubation: Incubated at room temperature for 1 hour. Antibody preparation (1:2000 dilution): 7.5 μl of antibody was pipetted and 15 ml of primary antibody (anti-VZV gE protein antibody, mouse mAb) dilution solution was added.

[0310] Film washing: 1x TBST buffer was used as the washing solution, and the film was washed three times for 5 minutes each time. Secondary antibody incubation: Incubated at room temperature for 1 hour. Antibody preparation (1:1000 dilution): 30 μl of antibody was pipetted and 30 ml of secondary antibody (goat anti-mouse IgG) diluted solution was added. Film washing: 1x TBST buffer diluted with distilled water was used as the washing solution, and the film was washed three times for 5 minutes each time. Color development: 1 ml each of developer and fixer were mixed in a 1:1 ratio, and added dropwise to the film to develop the color, and then the film was photographed.

[0311] Internal reference protein GAPDH: The relative intensity was consistent, indicating that the loading amounts of different samples were the same. Based on this, the expression intensity between different samples was compared. Negative control NC: A band corresponding to the internal reference region was present only, and no corresponding band was present in the target protein region, fulfilling the requirements of a negative control. Positive control PC (gE purified protein): A corresponding band was present within the target size and in the internal reference region, fulfilling the requirements of a positive control. Under conditions where the negative and positive settings were reasonable and the bands were clear, the presence or absence of expression in the experimental sample and the relative expression level were determined. The Western blot results of the protein sample mixture obtained by transfecting cells with the series of mRNA vaccines prepared in Example 2 and then lysing them are shown in Figure 2 herein.

[0312] Example 4: Immunization of Balb / c mice Experimental materials Animals: Balb / c mice, female, 6-8 weeks old. Test articles: 1) mRNA vaccine corresponding to the antigen expressed in Example 3, 2) LNP sample (mRNA-free LNP sample prepared in Example 2), 3) GSK recombinant subunit vaccine (Shingrix®), Grouping: Mice were randomly divided into groups according to body weight, with 5 mice in each group.

[0313] Animal immunization: Mice were immunized with the vehicle or test article (liquid mRNA vaccine prepared in Example 2) on days 0 and 28, respectively, by intramuscular injection into the gastrocnemius muscle, with the volume and dosage as shown in the table below.

[0314] [Table 7] TIFF2025179041000027.tif32169

[0315] [Table 8] TIFF2025179041000029.tif25169

[0316] Sample collection and preparation Approximately 0.2 ml of blood was collected from the orbital venous plexus of each animal, and the serum was separated (by centrifugation at 4000 rpm / min for 10 minutes at 4°C) and stored in a refrigerator at 4°C until processing.

[0317] Health Monitoring Twice daily (once in the morning and once in the afternoon) and include, but are not limited to, mortality, morbidity, respiratory discharge, secretions, feces, and eating and drinking habits.

[0318] Humane Endpoint According to IACUC protocol, mice that lost more than 20% of their body weight during the experiment (before infection, the weight on day 0 was used as the baseline weight, and after infection, the weight on the day of inoculation was used as the baseline weight) and / or showed signs of moribundity were euthanized and recorded as dead animals in the results.

[0319] Example 5: Detection of gE protein-specific IgG antibody titers in Balb / c mouse serum by enzyme-linked immunosorbent assay (ELISA) Main experimental materials and suppliers The Varicella-Zoster Virus (VZV) IgG (human) titer ELISA detection kit (gE protein) was purchased from Acrobiosystems Co., Ltd. The solutions included in the kit include 1x washing buffer, positive control working solution, negative control working solution, dilution buffer, HRP-goat anti-mouse IgG, substrate solution, stop solution, and ELISA plates.

[0320] Experimental steps Preparation of working solutions: Preparation of 1x washing buffer: Take 50 ml of 10x washing buffer and dilute it to a volume of 500 ml with ultrapure water or deionized water. A positive control working solution and a negative control working solution were prepared.

[0321] Test sample pretreatment: Serial blood samples obtained in Example 4 on days 42 and 56.

[0322] Antibody titer test: The test sample, positive control PC, and negative control NC were diluted with dilution buffer at 1:100 to 1:102400.

[0323] Numbering: Dilutions were numbered to correspond to the wells of the ELISA plate, and PC and NC working solutions were set for each experiment.

[0324] Addition of samples: First, 100 μl of diluted samples, positive control working solution, and negative control working solution were added to the corresponding wells of the plate, shaken to mix evenly, and then incubated at 37°C for 1.0 hour.

[0325] Washing the plate: Discard the liquid in the wells, tap the ELISA plate dry, wash the plate with 1x washing buffer, soak 300 μl / well for 30 seconds, tap the plate, wash the plate three times in total, and tap dry.

[0326] Addition of HRP enzyme-labeled substance: HRP-goat anti-mouse IgG was diluted 1000-fold with dilution buffer, and 100 μl of it was added to each well, followed by incubation at 37° C. for 1.0 hour.

[0327] Washing the plate: Repeat step 6 to wash the plate three times.

[0328] Color development: 100 μl of substrate solution was added to each well and incubated in the dark at 37° C. for 20 minutes.

[0329] Stop: 50 μl of stop solution was added to each well and the ELISA plate was shaken until evenly mixed.

[0330] Data reading: Use a microplate reader to measure the OD 450 nm and OD 630 The absorbance value of each well was measured at wavelengths of 630 nm, and the background was set at 630 nm. and read the data within three minutes.

[0331] Interpretation of test results: Antibody positive: OD450 nm-OD 630 nm ≥ 0.1.

[0332] Example 6: CD4 secreting IFN-γ and IL-2 in the spleen of Balb / c mice by multiparameter flow cytometry (FCM) + T cells and CD8 + Detection of T cell percentage The action of immune checkpoint inhibitors is to prevent T cell exhaustion. We evaluated immune cell function by detecting cytokine production in tumor samples. When inflammation or host immune defense occurs, T cells and NK cells execute the immune response by producing gamma interferon (IFN-γ). Multiparameter flow cytometry (FCM) was used to detect IFN-γ-producing CD8+ cells in mouse spleens. + or CD4 + T cells were quantitatively analyzed.

[0333] Gamma interferon (IFN-γ) is a soluble dimeric cytokine and the only member of the type II interferon family. It is primarily secreted by natural killer cells (NK) and natural killer T cells (NKT) and plays a role in innate immunity, as well as by CD4 Th1 and CD8 cytotoxic T cells in the process of antigen-specific immunity. IFN-γ, or type II interferon, plays an important role in innate and adaptive immunity against viral, certain bacterial, and protozoan infections. IFN-γ is an important activator of macrophages and an inducer of major histocompatibility complex type II (MHC II) expression. Interleukin-2 (IL-2) is a cytokine in the chemokine family. It is a cytokine derived from multiple cells (mainly produced by activated T cells) with pleiotropic effects (mainly promoting the growth, proliferation, and differentiation of lymphocytes). It plays an important role in the body's immune response and anti-viral infection, stimulating the proliferation of T cells activated by specific antigens or mitogenic factors, activating T cells, promoting cytokine production, stimulating the proliferation of NK cells, enhancing NK killing activity and cytokine production, inducing the production of lymphokine-activated killer cells (LAK), promoting B cell proliferation and antibody secretion, and activating macrophages.

[0334] Detection method: FCM (multiparameter flow cytometry).

[0335] Detection sample: spleen removed on day 56 of the immunized mouse model of Example 5.

[0336] Detection objective: CD4 secreting IFN-γ and IL-2 in mouse spleen + T cells and CD8 + The goal is to measure the proportion of T cells.

[0337] Detection indicators: CD4 + (IFN-γ + ): All CD4 that produce IFN-γ + T cell number, CD4 + (IL-2 + ): All CD4 that produce IL-2 + T cell number, CD4 + (IFN-γ + IL-2 + ): CD4 that simultaneously produces IFN-γ and IL-2 + T cell number, CD4 + (IFN-γ + or IL-2 + ): All CD4 cells that produce IFN-γ or IL-2 + T cell number, CD8 + (IFN-γ + ): All CD8 that produce IFN-γ + T cell number, CD8 + (IL-2 + ): All CD8 that produce IL-2 + T cell number, CD8 + (IFN-γ + IL-2 + ): CD8 that simultaneously produces IFN-γ and IL-2 + T cell number, CD8 + (IFN-γ + or IL-2 + ): All that produce IFN-γ or IL-2 CD8 + T cell count.

[0338] Experimental materials Key Reagents and Suppliers

[0339] [Table 9] TIFF2025179041000031.tif48169

[0340] Preparation of reagents Each reagent was prepared in the amount required for the experiment, and the preparation ratio of the reagents was a volume ratio.

[0341] FBS inactivation: FBS stored at -20°C was left to dissolve at 4°C, inactivated by placing in a 56°C water bath for 30 minutes, filtered through a 0.22 μm filter, and stored at 2-8°C. The expiration date was 14 days.

[0342] Preparation of 1x RBC lysis buffer: 10x RBC lysis buffer (Mμlti-species) was diluted with sterile water to 1x RBC lysis buffer, mixed evenly and stored.

[0343] Preparation of complete medium containing 1% double antibody: Complete medium was prepared according to the following formula: RPMI-1640:inactivated FBS=9:1.

[0344] Complete medium containing 1% double antibody was prepared according to the following ratio: complete medium: penicillin-streptomycin mixture = 99:1. After homogeneous mixing, it was stored at 2-8 °C with a shelf life of 14 days. It was allowed to equilibrate to room temperature before use.

[0345] PMA / ionomycin positive stimulator working solution: Take 400 μl of PMA working solution and 80 μl of ionomycin (ION) and add them to 19.520 ml of complete medium containing 1% double antibody to prepare the positive stimulator working solution. The final concentration of PMA was 1 μg / ml, and the final concentration of ion was 2 μg / ml.

[0346] Negative control: complete medium containing 1% double antibody.

[0347] Preparation of peptide working solution: Protein transport inhibitor working solution, stock solution was diluted 50-fold with complete medium containing 1% double antibody.

[0348] Live / Dead Cell Working Solution: Zombie Violet TM The Fixable Viability solution was diluted 1000-fold with 1x PBS.

[0349] Surface staining antibody: For example, prepare the surface staining antibody for one well according to the ratio in the table below, mix thoroughly by pipetting, and store at 2-8°C in the dark for future use.

[0350] [Table 10]

[0351] 1x Perm / Wash Buffer: 10x Perm / Wash Buffer was diluted 10 times with ultrapure water.

[0352] Intracellular staining antibody: Using the dosage per well as an example, prepare the intracellular staining antibody according to the ratio in the table below, mix thoroughly by pipetting, and store at 2-8°C in the dark for future use.

[0353] [Table 11]

[0354] Preparation of cryopreservation solution: FBS:DMSO=9:1 was prepared and used immediately.

[0355] Operation steps Isolation of mouse splenocytes The cell strainer was placed on a 6-well plate containing 4 ml of complete medium containing 1% double antibody. The spleen alone was then placed on the cell strainer. Using the plunger end of a syringe, the spleen was gently crushed or pulverized on the cell strainer. The resulting cell suspension was passed through the cell strainer and filtered into a 15 ml centrifuge tube. 1–2 ml of complete medium containing 1% double antibody was added to wash the 6-well plate. The resulting liquid was then filtered and placed into a 15 ml centrifuge tube. The plate was centrifuged at 500 xg for 5 minutes at 20°C. The supernatant was discarded.

[0356] Cells were resuspended in 2 ml of 1x RBC lysis buffer and lysed for 5 ± 1 min. Centrifuged at 500 g for 5 min at 20°C. The supernatant was discarded. Cells were washed with 5 ml of complete medium containing 1% double antibody and centrifuged at 500 g for 5 min at 20°C. The supernatant was discarded.

[0357] The cells were resuspended in 10 ml of complete medium containing 1% double antibody and mixed evenly. Immediately, 100 μl of the cell suspension was transferred to an EP tube. Next, 10 μl of the cell suspension and 10 μl of fluorescent staining solution were transferred to an EP tube and mixed evenly. After that, 10 μl was taken and counted in a fluorescence counter to calculate the total number of live cells and cell viability.

[0358] Centrifuge the cells in the 15 ml centrifuge tubes at 400 g for 5 min at 20°C, discard the supernatant, and resuspend the cells in fresh complete medium containing 1% double antibody to a viable cell concentration of 1 × 10 for each sample. 7 Adjusted to cells / ml.

[0359] After the sample was applied to the plate, the remaining cells were centrifuged at 400 g for 5 min at 20°C, and the cell concentration was adjusted to (1–2) × 10 using a cryopreservation solution. 7 The cells were adjusted to cells / ml, dispensed into pre-marked cryopreservation tubes at 1 ml / tube, placed in a programmed cooling box, frozen in a refrigerator at -70°C or below, and transferred to a liquid nitrogen tank for long-term storage after 12–24 hours.

[0360] Stimulation of mouse splenocytes Stimulation of mouse splenocytes was performed in 96-well U-shaped plates, and the day of the experiment was marked on the plates.

[0361] Positive, negative, and polypeptide wells were set up according to the experimental content: 100 μl / well of negative control was added to the negative wells, 100 μl / well of positive stimulant working solution was added to the positive wells, 100 μl / well of polypeptide working solution was added to the peptide wells, and then 100 μl / well of cell suspension was added to the positive, negative, and peptide wells.

[0362] The 96-well plate was placed in a 37°C, 5% CO2 incubator and stimulated for 2 hours ± 5 minutes, after which 10 μl / well of protein transport inhibitor working solution was added and mixed evenly, followed by placing in a 37°C, 5% CO2 incubator for 18 hours ± 10 minutes of stimulation, after which the round-bottom 96-well plate was removed and flow staining was performed.

[0363] NOTE: The experimental procedures in this section must be performed in a biological safety cabinet, and all reagents were sterile.

[0364] Staining of cells Washing of samples after stimulation: The round-bottom 96-well plate was removed and centrifuged at 500 g for 5 minutes at 4°C, and the supernatant was discarded. 200 μl of PBS was added to each well, resuspended, mixed evenly, centrifuged at 500 g for 5 minutes at 4°C, and the supernatant was discarded.

[0365] Live / dead cell staining: 50 μl of the prepared live / dead cell working solution was added to each well, resuspended, mixed evenly, and stained in the dark at 2°C to 8°C for 30 minutes. After staining, 150 μl of PBS was added to each well, and the mixture was centrifuged at 500 g and 4°C for 5 minutes, and the supernatant was discarded.

[0366] Surface staining: 50 μl of surface staining antibody was added to each well, resuspended, mixed evenly, and stained in the dark at 2°C to 8°C for 30 minutes. Staining was stopped by adding 150 μl of cell staining buffer per well, followed by centrifugation at 500 g for 5 minutes at 4°C, and the supernatant was discarded.

[0367] Cell fixation and membrane disruption: 100 μl / well of fixation and permeabilization solution was added, resuspended, mixed evenly, and incubated in the dark at 4°C for 20 minutes. 100 μl / well of 1×Perm / Wash buffer was added to stop the cells, and the cells were centrifuged at 500 g for 5 minutes at 4°C, and the supernatant was discarded.

[0368] Intracellular staining: 50 μl of intracellular staining antibody was added to each well, resuspended, mixed evenly, and incubated in the dark at 4°C for 50 minutes. Staining was stopped by adding 150 μl / well of 1×Perm / Wash buffer, centrifuged at 500 g for 5 minutes at 4°C, and the supernatant was discarded. 200 μl / well of 1×Perm / Wash buffer was added, resuspended, mixed evenly, centrifuged at 500 g for 5 minutes at 4°C, and the supernatant was discarded.

[0369] Filtration: Add 200 μl / well of cell staining buffer, resuspend, mix evenly, and filter the cell suspension into each new well plate using a 300-mesh nylon mesh. An example of the loading number is "01-P-1", where "01" represents the sample serial number, P represents the positive well, and the last 1 represents the well serial number.

[0370] Instrumental detection Device name: Flow cytometer (Cytoflex), Instrument collection parameters: collection flow rate was set to fast, and the number of records was 100,000 cells collected under the lymphocyte gate.

[0371] Data Processing Data Analysis: CD3 + Circle the cells, then CD4 + Cells and CD8 + Each cell is circled. CD4 + CD4 from each cell + (IFN-γ + ) subgroups and CD4 + (IL-2 + ) subgroups are circled, CD8 + CD8 from each cell + (IFN-γ + ) subgroups and CD8 + (IL-2 + ) Subgroups are circled.

[0372] Data calculation formula: All values ​​of this detection were statistically expressed as percentages, and the valid value was calculated as the average value of polypeptide wells minus the average value of negative wells. All values ​​were rounded to two decimal places.

[0373] Example 7: Detection of IFN-γ and IL-2 cytokines secreted by spleen cells of Balb / c mice by enzyme-linked immunospot technique (ELISPOT) Enzyme-linked immunospot assay (ELISPOT) is a highly sensitive detection method for cellular immunology research, capable of detecting antibody-secreting cells (ASCs) and cytokine-secreting cells (CKs) at the single-cell level. It is more sensitive than ELISA and limiting dilution assays, being able to detect a single protein-secreting cell from 200,000 to 300,000 cells. Furthermore, this method can perform functional detection on live cells after antigen stimulation. Because of its high specificity, intuitive reliability, and ease of operation, it has been widely used in the immunology community both at home and abroad to detect secreted CK cells and measure ASCs. After antigen stimulation, lymphocytes locally produce cytokines, which are captured by specific monoclonal antibodies on a pre-coated PVDF membrane at the bottom of the ELISPOT plate. After cell removal, the captured cytokines were bound to biotin-labeled monoclonal antibodies, followed by alkaline phosphatase- or horseradish peroxidase-labeled avidin. After adding a substrate to develop color, purple or reddish-brown spots appeared on the PVDF membrane, indicating that the cells produced cytokines. The spots were automatically counted and analyzed using an enzyme-linked immunospot assay (ELISPOT Reader).

[0374] The spleen organs isolated from the mouse model of Example 6 were used to detect the contents of IFN-γ and IL-2 secreted by spleen cells stimulated with the peptide library on day 56 after immunization.

[0375] Sample processing: Mouse spleen cell isolation and cryopreservation process.

[0376] Elispot method: The contents of IFN-γ and IL-2 in the supernatant of splenocytes after stimulation with the peptide library were detected.

[0377] Main Reagents

[0378] [Table 12]

[0379] Preparation of reagents 1) FBS inactivation: FBS stored at -20°C was thawed at 4°C and inactivated in a 56°C water bath for 30 minutes. It was then filtered through a 0.22 μm filter and stored at 2-8°C. The expiration date was 30 days.

[0380] 2) Preparation of PMA working solution: Add 1 ml of DMSO to the PMA reagent bottle to dissolve the PMA powder, then transfer it to a 15 ml centrifuge tube. Rinse the PMA reagent bottle with 1 ml of DMSO and combine the rinse solution into the 15 ml centrifuge tube. Add 8 ml of DMSO to the 15 ml centrifuge tube and mix thoroughly to prepare a PMA working solution with a concentration of 100 μg / ml. Store in a dark place in a refrigerator at -20°C.

[0381] 3) Preparation of ION working solution: Add 2 ml of DMSO to the ION reagent bottle to dissolve the ION powder, then transfer it to a 15 ml centrifuge tube and use 1 ml of DMSO to dissolve the ION powder. The N reagent bottle was rinsed, and the rinsing step was repeated twice (a total of three times, 1 ml each time). The rinse solution was combined into a 15 ml centrifuge tube, and 5 ml of DMSO was added to the 15 ml centrifuge tube and mixed evenly to prepare an ION working solution with a concentration of 1 mg / ml. This was then stored in a refrigerator at -20°C.

[0382] 4) Preparation of complete medium containing 1% double antibody: Complete medium was prepared according to the following ratio: RPMI-1640:inactivated FBS = 9:1. Complete medium containing 1% double antibody was prepared according to the following ratio: complete medium:penicillin-streptomycin mixture = 99:1. After uniform mixing, it was stored at 2-8°C with a shelf life of 14 days. It was allowed to equilibrate to room temperature before use.

[0383] 5) Preparation of positive stimulator working solution: Take 400 μl of PMA working solution and 80 μl of ionomycin (ION) and add them to 19.520 ml of complete medium to prepare the positive stimulator working solution. The final concentration of PMA was 2 μg / ml, and the final concentration of ION was 4 μg / ml.

[0384] 6) Preparation of polypeptide working solution: (1) Preparation of gE peptide library solution: To each tube of lyophilized powder, 100 μl of DMSO was added, and after dissolution, the polypeptide concentration in each tube was 1 mg / ml / peptide. 900 μl of PBS was added to the dissolved peptide solution, and the gE peptide library concentration was adjusted to 100 μg / ml / peptide.

[0385] (2) Preparation of gE working solution: The gE peptide library solution was prepared according to the following ratio: 1:50: complete medium containing 1% double antibody. After uniform mixing, the concentration of the gE working solution was 2 μg / ml / peptide.

[0386] Negative Control: The negative control was complete medium containing 1% double antibody.

[0387] 7) Preparation of 1x PBS solution: Prepare according to the following: 20x PBS: pure water = 1:19, store at room temperature, and use immediately after preparation.

[0388] 8) Preparation of PBS containing FBS: Prepare according to the following: PBS (Biotopped): inactivated FBS = 200:1 and use immediately after preparation.

[0389] 9) Preparation of antibodies: (1) Preparation of R4-6A2-biotin: R4-6A2-biotin: PBS containing FBS = 1:1000, prepared and used immediately.

[0390] (2) Preparation of 5H4-biotin: 5H4-biotin: PBS containing FBS = 1:1000, prepared and used immediately.

[0391] Experimental steps 1) Spleen organs were removed from the mouse model of Example 6, and on day 56 after immunization, the content of IFN-γ and IL-2 secreted by spleen cells stimulated with the peptide library and the CD4 and CD8 T cells producing IFN-γ and IL-2 were detected.

[0392] 2) Test sample: Spleen excised on day 56 and frozen and stored at -80°C.

[0393] Resuscitation of mouse splenocytes Preparation: Before resuscitation, adjust the temperature of the water bath to 37°C and prepare complete solution containing 1% double antibody. All media were pre-warmed to 37°C.

[0394] Sampling: The sample manager removed the mouse spleen cells frozen in the liquid nitrogen tank and transferred them to a box containing liquid nitrogen. The cryopreservation tubes were removed from the box, sprayed with 75% alcohol, and placed on the transfer window of the cell culture chamber.

[0395] Resuscitation: The validation staff immediately removed the cryopreservation tube from the transfer window and placed it in a 37°C water bath for rapid thawing. After the cells were fully resuscitated, they were transferred in a sterile environment to a 15 ml centrifuge tube containing 9 ml of complete medium containing 1% double antibody.

[0396] Rinse: 1 ml of complete medium containing 1% double antibody was added to the cryotube to rinse the cryotube, and the rinse was transferred to a 15 ml centrifuge tube.

[0397] Centrifugation: Centrifuged at 400 g for 10 minutes at 18-20°C.

[0398] Counting: The supernatant was discarded, and the cells were resuspended in 10 ml of complete medium containing 1% double antibody. After mixing evenly, 100 μl of the cell suspension was immediately transferred to an EP tube. 10 μl of the 100 μl cell suspension was immediately added to an EP tube containing 10 μl of fluorescent staining solution. After mixing evenly, 10 μl was taken and counted using a fluorescence counter. The total number of live cells and cell viability were calculated (if the viability was less than 50%, the count using the fluorescence counter was repeated once, and the second counting result was used as the reference).

[0399] Centrifugation again: The remaining cells in the 15 ml centrifuge tube were centrifuged at 400 g and 18-20°C for 5 minutes.

[0400] Resuspend: Discard the supernatant and resuspend the cells in complete medium containing 1% double antibody to a viable cell concentration of A0: 4 × 10 6 cells / ml, A1:4×10 5 Adjusted to cells / ml.

[0401] Stimulation of mouse splenocytes Washing of plates: IFN-γ and IL-2 well plates were taken out, and 200 μl / well of PBS (PBS used throughout the experiment was sterile) was added and washed four times.

[0402] Blocking: Discard the PBS in the wells, tap the plate (to remove as much remaining liquid as possible from the wells), add 200 μl / well of complete medium containing 1% double antibody, and leave at room temperature for at least 30 minutes (the specific time was recorded honestly).

[0403] Plating: Discard the complete medium containing 1% double antibody in the well plate, tap the plate (to remove as much remaining liquid in the well as possible), add 100 μl / well of positive stimulant working solution to the positive wells, add 100 μl / well of negative control product to the negative control wells, add 100 μl / well of polypeptide working solution to the polypeptide wells, and blank wells contain no cells; add only 200 μl of negative control product to each well.

[0404] Cell addition: A0 cell suspension was added to the negative and polypeptide wells at 100 μl / well, and A1 cell suspension was added to the positive wells at 100 μl / well.

[0405] Culture: The plates were wrapped in aluminum foil and placed in a 37°C, 5% CO2 incubator for 21 hours ± 30 minutes of stimulation.

[0406] NOTE: The experimental procedures in this section must be performed in a biological safety cabinet. All drugs were sterile reagents.

[0407] Cytokine detection Washing of the plate: The next day, the original liquid in the well plate was emptied, the plate was tapped (to remove as much remaining liquid in the well as possible), 200 μl / well of PBS was added, and the plate was washed five times.

[0408] Addition of secondary antibody: Discard the PBS in the wells and tap the plate (to remove as much remaining liquid as possible from the wells). Add 100 μl / well of IFN-γ:R4-6A2-biotin working solution. Add 100 μl / well of IL-2:5H4-biotin working solution. Incubate at room temperature for 2 hours ± 5 minutes.

[0409] Washing of the plate: The original liquid in the well plate was emptied, the plate was tapped (to remove as much remaining liquid in the well as possible), 200 μl / well of PBS was added, and the plate was washed five times.

[0410] Addition of tertiary antibody: Streptavidin-HRP working solution was added at 100 μl / well. After incubation at room temperature for 1 hour ± 2 minutes, the liquid in the well plate was emptied, the plate was tapped (to remove as much liquid as possible from the well), PBS was added at 200 μl / well, and the plate was washed five times.

[0411] Washing of the plate: The original liquid in the well plate was emptied, the plate was tapped (to remove as much remaining liquid in the well as possible), 200 μl / well of PBS was added, and the plate was washed five times.

[0412] Color development: 100 μl of TMB substrate solution was added to each well after bringing it to room temperature. After allowing the color to develop at room temperature for 15±1 minutes, the solution in the well plate was discarded and the well plate was washed with pure water to stop the color development. The bottom plate of the well plate was removed and the bottom of the well plate was rinsed.

[0413] Data reading: After the well plate was completely dried in the dark at room temperature, a photograph was taken using an ELISpot analyzer and the data was read. Camera parameters: shutter (exposure) (exposure intensity): 400, gain (gain value): 25. Counting parameters: IFN-γ-mouse-Mab / IL-2-mouse-MabT: size: 30, intensity (optical density threshold): 30, TNTC: 80%.

[0414] Example 8: VZV gE antigen distribution in African green monkey kidney cells (Vero) Vero cells are a cell line used for cell culture. The "Vero" line is isolated from kidney epithelial cells extracted from African green monkeys. Vero cells were transfected with various mutant gE antigens (YK-VZV-010, YK-VZV-011, YK-VZV-013, YK-VZV-018, and YK-VZV-020) constructed by our company. The transfected cells were stained with antibodies for gE antigen labeling and Golgi marker GM130 / TGN46 antibodies, and the localization of the antigens was observed using a confocal microscope. The results are shown in Figures 3 and 4.

[0415] Preparation of reagents and consumables

[0416] [Table 13] TIFF2025179041000036.tif67169

[0417] Experimental steps 1) Cell seeding: Vero cells (purchased from Beyotime Biotechnology) were seeded on a small confocal dish. The number of cells seeded was recorded when the cell seeding density reached approximately 30% under a microscope. The cells were cultured overnight.

[0418] 2) Transfection: Cells were transfected with the overexpression stock solution (50 ng / well) and cultured for 48 hours.

[0419] 3) Washing: Remove the confocal dish, remove the medium directly, and add PBS to wash 1 to 3 times, taking care not to shake vigorously during washing.

[0420] 4) Fixation: Cells were fixed by adding 200 μl of 4% paraformaldehyde solution, left at room temperature for 15 minutes, and washed three times with 1×PBS, each time left at room temperature for 5 minutes.

[0421] 5) Permeabilization: 200 μl of 0.3% Tritonx-100 (diluted in 1× PBS) was added to the wells for 10 minutes, followed by washing three times with 1× PBS, each time leaving the wells at room temperature for 5 minutes.

[0422] 6) Blocking: 200 μl of immunostaining blocking solution was added for blocking, and the blocking was carried out at room temperature for 2 hours.

[0423] 7) Primary antibody binding: Add 100 μl of primary antibody diluted with antibody diluent (VZV gE antibody 1:100, 647-conjugated GOLGA2 / GM130 antibody 1:100, CoraLite® 594-conjugated TGN46 antibody 1:100). Incubate overnight at 4°C in the dark. Remove the primary antibody by aspiration (it can be recycled).

[0424] 8) Washing of primary antibody: Wash with 1x PBS three times for 5 minutes each time to remove unbound primary antibody.

[0425] 9) Secondary antibody incubation: 200 μl of fluorescent secondary antibody diluted with antibody diluent was added, the antibody dilution ratio was 1:300, and the cells were incubated in the dark at room temperature for 1.5 hours.

[0426] 10) Secondary antibody washing: To avoid non-specific staining, wash with 1x PBS three times for 5 minutes each time to remove unbound secondary antibody.

[0427] 11) Nuclear staining and sealing: Remove the frame, add one drop of DAPI staining stock solution to each well, cover, and store in the dark at 4°C.

[0428] 12) Photographs were taken and observed using a confocal microscope.

[0429] Control settings The control settings of the present invention are as follows. LNP: blank LNP containing no mRNA vaccine; Shingrix®: Shingrix® is a recombinant protein subunit vaccine prepared and currently marketed by GSK Plc. It is the extracellular domain of the VZV gE protein, amino acids 1-539, and has no mutations.

[0430] Comparative Example 1 Based on the literature (Morgan A, et al. 2020) and the Moderna patent (US11643441B1) reporting the varicella zoster mRNA vaccine antigen sequence, it was found that the antigen developed in this patent is gE(1-573aa)Y569A. Based on this, this project designed YK-VZV-004 (SEQ ID NO: 15), which maintains the same mutation site and shortened length as the varicella zoster mRNA vaccine antigen sequence gE(1-573aa)Y569A reported in the Moderna patent (US11643441B1). YK-VZV-004:gE(1-573aa)Y569A was used as a positive control for antigen screening to evaluate the immunogenicity of the vaccine along with the newly designed mutations and mutation combination sequences of the present invention. See the Examples for specific results.

[0431] Comparative Example 2 According to the varicella zoster mRNA vaccine antigen sequence reported in the patent (CN114081943A), the VZV gE full-length polypeptide developed in this patent was found to contain a combination of mutations (Y569A, S593A, S595A, T596A, T598A). Based on this, this project designed YK-VZV-006 (SEQ ID NO: 19), which maintained the same combination of mutation sites (Y569A, S593A, S595A, T596A, T598A) as SEQ ID NO: 2 described in the patent (CN114081943A). YK-VZV-006: gE (1-623aa), (Y569A, S593A, S595A, T596A, T598A) was also used as one of the antigen screening controls of the present invention to evaluate the immunogenicity of the vaccine together with the newly designed mutations and mutation combination sequences of the present invention, and specific results are shown in the Examples.

[0432] Comparative Example 3 Based on the varicella zoster mRNA vaccine antigen sequence reported in US20230233671A1, it was found that this patent discloses the mutant VZV gE protein Y582A. Based on this, this project designed YK-VZV-045 (SEQ ID NO: 171), which maintains the same mutation site combination Y582A as the mRNA vaccine antigen sequence described in US20230233671A1. YK-VZV-045 was also used as one of the antigen screening controls of the present invention to evaluate the immunogenicity of the vaccine together with the newly designed mutation and mutation combination sequences of the present invention. See the Examples for specific results.

[0433] Comparative Example 4 According to the varicella zoster mRNA vaccine antigen sequence reported in CN108472309A, it was found that this patent discloses the mutant VZV gE protein Y582A; Based on this, this project designed YK-VZV-007 (SEQ ID NO: 23), maintaining the same combination of mutation sites (Y582A, S593A, S595A, T596A, T598A) as the mRNA vaccine antigen sequence described in CN108472309A. YK-VZV-007 was also used as one of the antigen screening controls of the present invention to evaluate the immunogenicity of the vaccine together with the newly designed mutations and mutation combination sequences of the present invention. See the Examples for specific results.

[0434] Summary and analysis of results Results of in vitro expression of VZV gE antigen mutants The Western blot detection results of the 43 VZV gE antigen mutants (Fig. 2) showed that the expression of the YK-VZV-002 antigen protein was extremely low or almost nonexistent, and the expression results of the YK-VZV-041, YK-VZV-042, and YK-VZV-043 antigen proteins were negative, meaning no expression. Therefore, the above four sequences cannot be used as candidate antigens for varicella-zoster mRNA vaccines and were therefore excluded.

[0435] All other antigen variants showed clear protein expression, and the size of the expressed proteins was approximately 63 kDa to 75 kDa, which was consistent with the theoretical design of the antigen and could be used as effective candidate antigens for the next step of immunogenicity screening and evaluation.

[0436] From the results of the above Western blot experiments, YK-VZV-001, YK-VZV-003, YK-VZV-004 (Comparative Example 1), YK-VZV-006 (Comparative Example 2), YK-VZV-007 (Comparative Example 4), YK-VZV-009, YK-VZV-010, YK-VZV-011, YK-VZV-012, YK-VZV-013, YK-VZV-014, YK-VZV-015, YK-VZV-016, YK-VZV-017, YK-VZV-018, YK-VZV-019, YK-VZV-020, YK-VZV-021, YK-VZV-022, YK-VZV-023, YK-VZV-024, YK-VZV-025, YK-VZV-026, YK-VZV-027, YK-VZV-028, YK-VZV-029, YK-VZV-030, YK-VZV-031, YK-VZV-032, YK-VZV-033, YK-VZV-034, YK-VZV-035, YK-VZV-036, YK-VZV-037, YK-VZV-038, YK-VZV-039, YK-VZV-040, YK-VZV-041, YK-VZV-042, YK-VZV-043, YK-VZV-044, YK-VZV-045, YK-VZV-046, YK-VZV V-013, YK-VZV-014, YK-VZV-015, YK-VZV-016, YK-VZV-017, YK-VZV-018, YK-VZV-019, YK-VZV-020, YK-VZV-021, YK-VZV-022, YK-VZV-023, YK-VZV-024, YK-VZV-025, YK-VZV The above 39 antigen sequences, YK-VZV-026, YK-VZV-027, YK-VZV-028, YK-VZV-029, YK-VZV-030, YK-VZV-031, YK-VZV-032, YK-VZV-033, YK-VZV-034, YK-VZV-035, YK-VZV-036, YK-VZV-037, YK-VZV-038, YK-VZV-039, YK-VZV-040, YK-VZV-044, and YK-VZV-045 (Comparative Example 3), all had significant protein expression and could be used as effective candidate antigens for the next step of immunogenicity screening and evaluation. Therefore, gE-specific antibody IgG titer detection was performed against the above antigens.

[0437] Results of gE protein-specific IgG antibody titers of VZV gE antigen mutants in Balb / c mouse sera (D42) As can be seen from Example 5, the gE protein-specific IgG antibody titer in the serum of mice 42 days after immunization was detected by ELISA, and the detection results are shown in Table 11 below.

[0438] [Table 14] TIFF2025179041000038.tif224168TIFF2025179041000039.tif215168TIFF2025179041000040.tif92168

[0439] Results: From the gE-specific antibody IgG titer results on day 42, the titer result of the Shingrix® positive control product was 190 x 10 4 Among them, the variants YK-VZV-009, YK-VZV-011, YK-VZV-013, YK-VZV-014, YK-VZV-010, YK-VZV-012, YK-VZV-018, YK-VZV-020, YK-VZV-030, YK-VZV-031, YK-VZV-038, YK-VZV-021, and YK-VZV-024 are more effective than Shingrix®. , YK-VZV-028, YK-VZV-003, YK-VZV-001, YK-VZV-016, YK-VZV-015, YK-VZV-017 and YK-VZV-004 (Comparative Example 1), YK-VZV-006 (Comparative Example 2), YK-VZV-045 (Comparative Example 3), YK-VZV-007 (Comparative Example 4), and the efficacy was shown to be 2.7 to 5.3 times that of the Shingrix (registered trademark) positive control.

[0440] Based on the fact that the gE-specific antibody IgG titer was 1.8 to 2.8 times higher than that of the YK-VZV-007 sequence (Comparative Example 4) using the Shingrix® positive control as a standard, the following 14 mutants, YK-VZV-009, YK-VZV-011, YK-VZV-013, YK-VZV-014, YK-VZV-010, YK-VZV-012, YK-VZV-018, and YK-VZV-019 were further evaluated. , YK-VZV-020, YK-VZV-030, YK-VZV-031, YK-VZV-038, YK-VZV-021, YK-VZV-024, YK-VZV-028 and four control sequences YK-VZV-004 (Comparative Example 1), YK-VZV-006 (Comparative Example 2), YK-VZV-045 (Comparative Example 3), and YK-VZV-007 (Comparative Example 4) were extracted and subsequent detection was performed.

[0441] Results of VZV gE antigen mutant cellular immunity (FCM) Cellular immunity, especially T cell immunity, plays an important role in the process of VZV infection, and research has shown that cell-mediated immune effects are an important index for evaluating the immune effect of varicella-zoster vaccines. To further evaluate the specific cellular immune response induced by immunizing mice with the above-mentioned antigen mutant mRNA, in Example 6, we used FCM to measure the specific CD4+ cells induced after immunizing mice with VZV gE antigen mRNA. + T cells and CD8 + The immune response of T cells was detected.

[0442] The Shingrix® positive control was used as a standard, and the gE-specific antibody IgG titer was as described above. Based on the fact that the effect was also superior to that of the YK-VZV-007 sequence, the following 14 variants were further selected: YK-VZV-009, YK-VZV-011, YK-VZV-013, YK-VZV-014, YK-VZV-010, YK-VZV-012, YK-VZV-018, YK-VZV-020, YK-VZV-030, YK-VZV-031, YK-VZV-038, YK-VZV-040, YK-VZV-041, YK-VZV-042, YK-VZV-043, YK-VZV-044, YK-VZV-045, YK-VZV-046, YK-VZV-047, YK-VZV-048, YK-VZV-049, YK-VZV-050, YK-VZV-051, YK-VZV-052, YK-VZV-053, YK-VZV-054, YK-VZV-055, YK-VZV-056, YK-VZV-057, YK-VZV-058, YK-VZV-059, YK-VZV-060, YK-VZV-061, YK-VZV-062, YK-VZV-063, YK-VZV-064, YK-VZV-065, YK-VZV-066, YK-VZV-067, YK-VZV-068, YK-VZV-069, YK-VZV-070, YK-VZV-071, Y The spleens of mice immunized with VZV-021, YK-VZV-024, YK-VZV-028, and four control sequences including YK-VZV-004 (Comparative Example 1), YK-VZV-006 (Comparative Example 2), YK-VZV-045 (Comparative Example 3), and YK-VZV-007 (Comparative Example 4) on day 56 were excised, and the CD4 receptors secreting IFN-γ and IL-2 in the mouse spleens were assayed using the detection method of Example 6. + T cells and CD8 + The percentage of T cells was detected.

[0443] CD4 + T cells CD4 + The immune effect of T cells plays an important role in the recovery from VZV infection and the immune effect of vaccines, and is one of the most important indicators for evaluating the immune effect of vaccines. Spleens were removed 56 days after immunization to measure the levels of IFN-γ and IL-2-secreting CD4 T cells in the spleens of mice. + The proportion of T cells was measured, and the specific results are shown in the table below.

[0444] [Table 15] TIFF2025179041000042.tif224168TIFF2025179041000043.tif57168

[0445] A) Based on the above results, the test products YK-VZV-010, YK-VZV-020, YK-VZV-013, YK-VZV-038, YK-VZV-012, YK-VZV-028, YK-VZV-031, YK-VZV-011, YK-VZV-021, YK-VZV-018, and YK-VZV-009 mRNA vaccines (11 in total) were screened to determine the secreted IFN-γ + CD4 + T cells and IL-2 + CD4 + The total percentage of T cells was more than twice as high as that of the Shingrix® positive control vaccine, up to 7 times higher, and 1.8 to 5.3 times higher than that of YK-VZV-007 (Comparative Example 4).It was also significantly superior to YK-VZV-004 (Comparative Example 1) and YK-VZV-045 (Comparative Example 3).

[0446] Among them, IFN-γ secreted by mRNA vaccines YK-VZV-010, YK-VZV-020, YK-VZV-013, YK-VZV-011, and YK-VZV-018 + CD4 + T cells and IL-2 + CD4 + The total percentage of T cells was 7.0, 5.2, 3.2, 2.7, and 2.3 times that of the Shingrix® positive control vaccine, and 5.3, 3.9, 2.4, 2.0, and 1.8 times that of YK-VZV-007 (Comparative Example 4), all of which were significantly higher than YK-VZV-004 (Comparative Example 1) and YK-VZV-045 (Comparative Example 3).

[0447] The specific information is shown in the table below.

[0448] [Table 16] TIFF2025179041000045.tif21169

[0449] B) However, the IFN-γ secreted by the mRNA vaccines tested, YK-VZV-014, YK-VZV-030, and YK-VZV-024, + CD4 + T cells and IL-2 + CD4 + The total percentage of T cells was less than twice that of the Shingrix® positive control vaccine and less than 1.5 times that of Comparative Example 4 (YK-VZV-007), so it was excluded, and the specific results are shown in the table below.

[0450] [Table 17]

[0451] IFN-γ + CD4 + T cell percentage A) Screening of the mRNA vaccines YK-VZV-010, YK-VZV-013, YK-VZV-011, YK-VZV-020, YK-VZV-012, YK-VZV-018, YK-VZV-028, YK-VZV-014, YK-VZV-038, and YK-VZV-031 was performed to determine the IFN-γ secreted by the mRNA vaccines. + CD4 + The percentage of T cells was optimal, ranging from 1.00 to 2.00%, significantly higher than in the comparative examples (YK-VZV-004, YK-VZV-045, YK-VZV-007), more than three times higher than the Shingrix® positive control vaccine, reaching a maximum of 6.7 times, and approximately 2.2 to 4.4 times higher than in comparative example 4 (YK-VZV-007).

[0452] Among them, IFN-γ secreted by mRNA vaccines corresponding to YK-VZV-010, YK-VZV-013, YK-VZV-011, YK-VZV-020, and YK-VZV-018 + CD4 +The percentage of T cells was 6.7, 5.7, 5.3, 5.3, and 4.0 times that of the Shingrix (registered trademark) positive control vaccine, and 4.4, 3.8, 3.6, 3.6, and 2.7 times that of Comparative Example 4 (YK-VZV-007), all of which were significantly higher than Comparative Example 1 (YK-VZV-004) and Comparative Example 3 (YK-VZV-045).

[0453] The specific results are shown in the table below.

[0454] [Table 18] TIFF2025179041000048.tif101169

[0455] B) IFN-γ secreted by mRNA vaccines YK-VZV-021, YK-VZV-009, YK-VZV-024, and YK-VZV-030 + CD4 + The T cell percentage was less than 1.0%, significantly lower than the cell percentage secreted by optimal and superior mRNA vaccines (1.0-2.0%), and only 2-3 times that of the Shingrix® positive control vaccine and approximately 1-2 times that of Comparative Example 4 (YK-VZV-007). Therefore, YK-VZV-021, YK-VZV-009, YK-VZV-024, and YK-VZV-030 were excluded from the candidate sequences, and the specific results are shown in the table below.

[0456] [Table 19]

[0457] IL-2 + CD4 + T cells A) Screening of mRNA vaccines corresponding to test articles YK-VZV-010, YK-VZV-020, YK-VZV-038, YK-VZV-031, YK-VZV-013, YK-VZV-028, YK-VZV-011, YK-VZV-012, YK-VZV-018, YK-VZV-021, and YK-VZV-009, and the IL-2 secreted by them + CD4 + The percentage of T cells was optimal, reaching 1.10-3.70%, which was significantly higher than that of the comparative mRNA vaccines (YK-VZV-004, YK-VZV-045, and YK-VZV-007), more than 3.7 times that of the Shingrix® positive control vaccine, and more than 2.2 times that of YK-VZV-007, and was significantly superior to Comparative Example 1 (YK-VZV-004) and Comparative Example 3 (YK-VZV-045).

[0458] Among them, IL-2 secreted by mRNA vaccines YK-VZV-010, YK-VZV-020, YK-VZV-013, YK-VZV-011, and YK-VZV-018 + CD4 + The total percentage of T cells was 12.3, 8.3, 4.3, 4.0, and 3.7 times that of the Shingrix® positive control vaccine, and 7.4, 5.0, 2.6, 2.4, and 2.2 times that of YK-VZV-007 (Comparative Example 4), all of which were significantly higher than YK-VZV-004 (Comparative Example 1) and YK-VZV-045 (Comparative Example 3).

[0459] The specific results are shown in the table below.

[0460] [Table 20] TIFF2025179041000051.tif16169

[0461] B) IL-2 secreted by the mRNA vaccines tested: YK-VZV-014, YK-VZV-030, and YK-VZV-024 + CD4 +The percentage of T cells was less than 1.0%, significantly lower than the cell percentage secreted by the optimal and superior mRNA vaccine (more than 1.0%), less than three times that of the Shingrix® positive control vaccine, and less than twice that of YK-VZV-007 (Comparative Example 4). Therefore, YK-VZV-014, YK-VZV-030, and YK-VZV-024 were excluded from the candidate sequences, and the specific results are shown in the table below.

[0462] [Table 21]

[0463] (4) Conclusion A) Percentage of immune cells in mouse spleens after immunization with mRNA vaccines of the screened sequences: IFN-γ + CD4 + T cells and IL-2 + CD4 + The total percentage of T cells was more than twice that of the Shingrix® positive control, reaching up to 7 times, and 1.8 to 5.3 times that of YK-VZV-007 (Comparative Example 4). FN-γ + CD4 + The percentage of T cells reached 1.00-2.00%, more than three times that of the Shingrix® positive control vaccine, and reached a maximum of 6.7 times, which was 2.2-4.4 times that of Comparative Example 4 (YK-VZV-007).

[0464] IL-2 + CD4 + The proportion of T cells was over 1% in both cases, more than 3.7 times that of the Shingrix® positive control vaccine and more than 2.2 times that of YK-VZV-007, and the information is as follows:

[0465] [Table 22] TIFF2025179041000054.tif207169

[0466] In the above sequence, CD4 induced by YK-VZV-010 + IFN-γ + T cells and CD4 + IL-2 + The proportion of T cells in spleen cells exceeded 2% in all cases, which was approximately 6 to 12 times higher than that of the Shingrix (registered trademark) positive control vaccine, demonstrating the best immune effect.

[0467] B) Excluded sequences are secreted by mRNA vaccines. IFN-γ + CD4 + T cells and IL-2 + CD4 + The total percentage of T cells is The efficacy was less than 2-fold that of the rix® positive control vaccine and less than 1.5-fold that of Comparative Example 4 (YK-VZV-007), Alternatively, IFN-γ secreted by mRNA vaccines + CD4 + The T cell count was less than 1.0%, which was only 2-3 times lower than the Shingrix® positive control vaccine and approximately 1-2 times lower than Comparative Example 4 (YK-VZV-007). Or IL-2 + CD4 + The percentage of T cells was less than 1.0%, less than three times that of the Shingrix® positive control vaccine and less than two times that of YK-VZV-007 (Comparative Example 4), the information of which is shown below.

[0468] [Table 23] TIFF2025179041000056.tif120170

[0469] CD4 levels of IFN-γ or IL-2 produced by the test mRNA vaccine + The proportion of T cells in spleen cells is less than 1%, and IFN-γ is also produced simultaneously. + CD4 + T cells and IL-2 + CD4 +The total proportion of T cells was less than 2.0 times that of Shingrix (registered trademark), the immunopotentiating effect was poor, and compared with the sequence of the above-mentioned A portion, it was not suitable as a candidate sequence for a varicella zoster vaccine.

[0470] In summary, the IFN-γ produced by the mRNA vaccines of the selected test substances YK-VZV-010, YK-VZV-013, YK-VZV-011, YK-VZV-020, YK-VZV-012, YK-VZV-018, YK-VZV-028, YK-VZV-038, and YK-VZV-031 was induced. + CD4 + T cells and IL-2 + CD4 + Since the effect of T cells was good, the above sequence and YK-VZV-004 (Comparative Example 1), YK-VZV-006 (Comparative Example 2), YK-VZV-045 (Comparative Example 3), and YK-VZV-007 (Comparative Example 4) were used in subsequent CD8 + Used for T cell immune analysis.

[0471] CD8 + T cells CD4 + Based on the results of T cell analysis, measurements were performed on test substances YK-VZV-010, YK-VZV-013, YK-VZV-011, YK-VZV-020, YK-VZV-012, YK-VZV-018, YK-VZV-028, YK-VZV-038, YK-VZV-031, and YK-VZV-004 (Comparative Example 1), YK-VZV-006 (Comparative Example 2), YK-VZV-045 (Comparative Example 3), and YK-VZV-007 (Comparative Example 4), and the CD8 T cells were assayed for the induction of IL-2 / IFN-γ produced by them. + The number of T cells was measured, and the specific results are shown in the table below.

[0472] [Table 24] TIFF2025179041000058.tif191170

[0473] IFN-γ +CD8 + T cells and IL-2 + CD8 + Total T cells A) Based on the above results, the secretion of IFN-γ was induced by the mRNA vaccines of the screened test products YK-VZV-010, YK-VZV-013, YK-VZV-020, YK-VZV-018, and YK-VZV-011. + CD8 + T cells and IL-2 + CD8 + The total percentage of T cells was 4 to 7 times higher than that of the Shingrix® positive control vaccine and approximately 3 to 5 times higher than that of YK-VZV-007 (Comparative Example 4), and the specific results are shown in the table below.

[0474] [Table 25]

[0475] B) However, the IFN-γ secreted by the mRNA vaccines tested, YK-VZV-012, YK-VZV-028, YK-VZV-038, and YK-VZV-031, + CD8 + T cells and IL-2 + CD8 + The total percentage of T cells was less than four times that of the Shingrix® positive control vaccine and less than three times that of YK-VZV-007 (Comparative Example 4), and the specific results are shown in the table below.

[0476] [Table 26]

[0477] IFN-γ + CD8 + T cells A) IFN-γ secreted by the mRNA vaccines tested: YK-VZV-010, YK-VZV-013, YK-VZV-020, YK-VZV-018, and YK-VZV-011 + CD8 +The T cell percentage was screened to be optimal, and all were above 5.0%, with YK-VZV-010 having the highest percentage at 8.10%, which was significantly higher than Comparative Examples 1 to 4 (YK-VZV-004, YK-VZV-006, YK-VZV-045, YK-VZV-007). The above five sequences were 4.5 to 7.0 times higher than the Shingrix (registered trademark) positive control vaccine and approximately 2.5 to 4.5 times higher than YK-VZV-007. The specific results are shown in the table below.

[0478] [Table 27]

[0479] B) IFN-γ secreted by the mRNA vaccines tested: YK-VZV-012, YK-VZV-028, YK-VZV-038, and YK-VZV-031 + CD8 + The percentage of T cells was less than 5%, significantly lower than the percentage of cells secreted by the optimal mRNA vaccine (5.00-8.10%), and was only 4.0 times or less than the Shingrix® positive control vaccine and approximately 1-2 times that of YK-VZV-007, so it was excluded. The specific results are shown in the table below.

[0480] [Table 28]

[0481] IL-2 + CD8 + T cells A) IL-2 secreted by the mRNA vaccines tested: YK-VZV-010, YK-VZV-013, YK-VZV-018, YK-VZV-011, YK-VZV-028, and YK-VZV-020 + CD8 +The optimal T cell percentage was found to be 0.3-0.5%, which was significantly higher than the comparative examples (YK-VZV-004, YK-VZV-006, YK-VZV-045, YK-VZV-007), more than 2.5-4.0 times that of the Shingrix (registered trademark) positive control vaccine, and approximately 1.5-3.0 times that of YK-VZV-007. The specific results are shown in the table below.

[0482] [Table 29]

[0483] B) IL-2 secreted by the mRNA vaccines tested: YK-VZV-038, YK-VZV-012, and YK-VZV-031 + CD8 + The proportion of T cells was only 0.2% or less, and the effect was not significantly different from or inferior to the Shingrix (registered trademark) positive control vaccine and YK-VZV-007, being only about 0.8 to 1.6 times that of the Shingrix (registered trademark) positive control vaccine and about 0.5 to 1.0 times that of YK-VZV-007. The specific results are shown in the table below.

[0484] [Table 30]

[0485] conclusion Percentage of splenic immune cells induced by the vaccine candidates screened in combination with the results of (1) to (3) above: A) IFN-γ + CD8 + T cells and IL-2 + CD8 + The total percentage of T cells was 4 to 7 times higher than that of the Shingrix® vaccine and approximately 3 to 5 times higher than that of YK-VZV-007 (Comparative Example 4). IFN-γ + CD8 +The percentage of T cells was optimal, being 5.0% or higher in all cases, 4.5 to 7.0 times higher than that of the Shingrix® positive control vaccine and approximately 2.5 to 4.5 times higher than that of YK-VZV-007. IL-2 + CD8 + The percentage of T cells was optimal, reaching 0.3-0.5%, which was 2.5-4.0 times higher than that of the Shingrix® positive control vaccine and approximately 1.5-3.0 times higher than that of YK-VZV-007. All of these results are significantly higher than those of the comparative examples (YK-VZV-004, YK-VZV-006, YK-VZV-045, YK-VZV-007). The information is as follows:

[0486] [Table 31] TIFF2025179041000066.tif183169

[0487] B) Percentage of mouse spleen immune cells induced by the test article vaccine cleared: IFN-γ + CD8 + T cells and IL-2 + CD8 + The total percentage of T cells was less than four times that of the Shingrix® vaccine and less than three times that of YK-VZV-007 (Comparative Example 4); Alternatively, IFN-γ + CD8 + The percentage of T cells was less than 5%, significantly lower than the percentage of cells secreted by the optimal mRNA vaccine (5.20-8.10%), only 4.0 times lower than the Shingrix® positive control vaccine, and approximately 1-2 times higher than YK-VZV-007. Or IL-2 + CD8 +The percentage of T cells was only 0.2% or less, and its effect was not significantly different or inferior to that of the Shingrix® positive control vaccine and YK-VZV-007, being only about 0.8 to 1.6 times that of the Shingrix® positive control vaccine and about 0.5 to 1.0 times that of YK-VZV-007, and therefore excluded. It was.

[0488] The information is as shown below.

[0489] [Table 32] TIFF2025179041000068.tif158169

[0490] IL-2 produced by YK-VZV-012 and YK-VZV-031 mRNA vaccines + CD8 + The percentage of T cells was lower than the effects of Shingrix® vaccine and YK-VZV-007 (Comparative Example 4), and therefore was excluded.

[0491] IFN-γ produced by YK-VZV-028 and YK-VZV-038 mRNA vaccines + CD8 + T cells and IFN-γ + and IL-2 + CD8 + The total percentage of T cells was significantly lower, only about twice that of the Shingrix® vaccine, and the sequences designed in the A portion (reaching more than seven times) had a lower immune effect and were not suitable as candidate sequences for a varicella zoster vaccine.

[0492] Therefore, based on the above results, the sequences with significant advantages in comprehensive immune effects include YK-VZV-010, YK-VZV-013, YK-VZV-011, YK-VZV-020, and YK-VZV-018, which have good immune effects and are significantly better than other sequences, among which YK-VZV-010 has the best effect.

[0493] Results of gE protein-specific IgG antibody titers of VZV gE antigen mutants in Balb / c mouse sera (D56) To verify the consistency of the above partial IgG antibody titers and partial VZV gE antigen variant cellular immunodetection results, the following partial test substances (YK-VZV-009, YK-VZV-010, YK-VZV-011, YK-VZV-013, YK-VZV-020, YK-VZV-014, YK-VZV-012, YK-VZV-018, YK-VZV-021, YK-VZV-024, YK-VZV-028, YK-VZV-029, YK-VZV-030, YK-VZV-031, YK-VZV-032, YK-VZV-033, YK-VZV-034, YK-VZV-035, YK-VZV-036, YK-VZV-037, YK-VZV-038, YK-VZV-039, YK-VZV-040, YK-VZV-041, YK-VZV-042, YK-VZV-043, YK-VZV-044, YK-VZV-045, YK-VZV-046, YK-VZV-047, YK-VZV-048, YK-VZV-049, YK-VZV-050, YK-VZV-051, YK-VZV-052, YK-VZV-053, YK-VZV-054, YK-VZV-055, YK-VZV-056, YK-VZV-057, YK-VZV-058, YK-VZV-059, YK-VZV-060, YK- The serum samples taken 56 days after immunization of mice were measured using the antibodies YK-VZV-030, YK-VZV-031, YK-VZV-038, and YK-VZV-004 (Comparative Example 1), YK-VZV-045 (Comparative Example 3), and YK-VZV-007 (Comparative Example 4). The VZV-binding-gE antibody IgG titer in the serum was detected using the detection method described in Example 5. The results are shown in the table below.

[0494] [Table 33] TIFF2025179041000070.tif193169

[0495] Based on the above results, on day 56, the gE-specific antibody IgG titers of the above mRNA vaccines were superior to the Shingrix® positive control, approximately 1.8 to 5.0 times that of the Shingrix® vaccine, and the antibody GMT was 270 to 750 × 10 4 Among them, the antibody GMTs of YK-VZV-010, YK-VZV-011, YK-VZV-018, YK-VZV-020, and YK-VZV-013 were 650 × 10 4 This was 4.0 to 5.0 times more effective than the Shingrix (registered trademark) vaccine and more than twice more effective than YK-VZV-007 (Comparative Example 4). Furthermore, it was significantly superior to YK-VZV-004 (Comparative Example 1) and YK-VZV-045 (Comparative Example 3).

[0496] The above IgG antibody titers and CD4 + T cells and CD8 + Based on the T cell detection results, five antigens, YK-VZV-010, YK-VZV-013, YK-VZV-011, YK-VZV-020, and YK-VZV-018, showed the best cellular immune effects, and were therefore used as candidate antigens for further detection and analysis by ELISpot.

[0497] VZV gE antigen variant cellular immune factor results (ELISOPT) ELISpot is the gold standard for screening and assessing the immune efficacy of candidate vaccine antigen-specific T cells and can be used to distinguish different subunits of activated T cells by cytokines, such as T helper cell (Th) type 1 cells (producing IFN-γ, IL-2, IL-6, IL-12, IL-21 and TNF-α cytokines), Th2 (producing IL-4, IL-5, IL-10 and IL-13 cytokines) and Th17 (producing IL-17 cytokine) cells.

[0498] Based on the combined IgG and cytokine detection results, five optimal antigen sequences (including YK-VZV-010, YK-VZV-013, YK-VZV-011, YK-VZV-020, and YK-VZV-018) were screened, and further detection was performed using the ELISpot method with Comparative Example 1 (YK-VZV-004), Comparative Example 2 (YK-VZV-006), Comparative Example 3 (YK-VZV-045), and Comparative Example 4 (YK-VZV-007), along with LNP and Shingrix® as controls. The results are as follows:

[0499] 1) The amount of cytokine IFN-γ secreted by the mRNA vaccines upon stimulation of T cells was 500 SFU or more in all cases, reaching a maximum of nearly 600 SFU, which was approximately 16 to 19 times that of the Shingrix (registered trademark) positive control and 2.5 to 3.0 times that of Comparative Example 4 (YK-VZV-007), and was significantly superior to the cytokines stimulated by the mRNA vaccines corresponding to Comparative Example 1 (YK-VZV-004), Comparative Example 2 (YK-VZV-006), or Comparative Example 3 (YK-VZV-045).

[0500] Among them, the mRNA vaccine prepared with the YK-VZV-010 sequence reached a maximum of 598.8 SFU, 18.7 times higher than the Shingrix® positive control and nearly 3 times higher than the YK-VZV-007 control.

[0501] The IFN-γ secretion levels were YK-VZV-010, YK-VZV-018, YK-VZV-011, YK-VZV-020, and YK-VZV-013, in order of highest to lowest. The differences between the groups were not significant, and the immune effects were all good and significant.

[0502] [Table 34]

[0503] 2) The cytokine IL-2 secretion levels of T cells stimulated by the mRNA vaccines were all 400 to 600 SFU or higher, 10 to 16 times higher than the Shingrix (registered trademark) positive control and 2.0 to 3.0 times higher than the YK-VZV-007 control, and were superior to the cytokine secretion stimulated by the mRNA vaccines corresponding to Comparative Example 1 (YK-VZV-004), Comparative Example 2 (YK-VZV-006), or Comparative Example 3 (YK-VZV-045).

[0504] Among them, the mRNA vaccine prepared with the YK-VZV-010 sequence reached a maximum of 614.5 SFU, 16.00 times higher than the Shingrix® positive control and more than three times higher than the YK-VZV-007 control.

[0505] The IL-2 secretion levels, from highest to lowest, were YK-VZV-010, YK-VZV-011, YK-VZV-018, YK-VZV-013, and YK-VZV-020, with no significant differences between groups, and the immune effects were all significant and favorable.

[0506] [Table 35]

[0507] Cellular localization results of VZV gE antigen mutants C-terminal truncation and "A 568 Y 569 R 570 V 571 " motif mutations reduced the localization of gE antigen to the transgolster network.

[0508] "A 568 Y 569 R 570 V 571 The " motif (SEQ ID NO: 184) is a transport motif that targets the gE polypeptide to the transglucan network. YK-VZV-018 (SEQ ID NO: 63) and YK-VZV-020 (SEQ ID NO: 71) encode truncated polypeptides (retaining 1-573 aa) in which the terminal 50 amino acids are deleted from the C-terminal region. 568 Y 569 R 570 V 571 Mutations were performed at any one position within the sequence (e.g., A568D and Y569K site mutations). The results of Example 8 showed that both the YK-VZV-018 and YK-VZV-020 mutants reduced the localization of gE polypeptide to the trans-Golgi network and enhanced its expression at the plasma membrane.

[0509] "A 568 Y 569 R 570 V 571 " or "Y 582 A 583 G 584 L 585Any combination of mutated motifs reduced the localization of gE antigen to the trans-Golgi network.

[0510] YK-VZV-011 (SEQ ID NO: 35) and YK-VZV-013 (SEQ ID NO: 43) encode the full-length gE polypeptide and are designated "A 568 Y 569 R 570 V 571 " motif (SEQ ID NO: 184), and further comprising the "Y 582 A 583 G 584 L 585 The endocytosis motif (coordinate number 185) was also mutated (e.g., Y582A). 568 Y 569 R 570 V 571 " or "Y 582 A 583 G 584 L 585 " Any combination of the mutant motifs showed that the gE antigen was expressed on the cell membrane.

[0511] 3) "A 593 E 594 A 595 A 596 D 597 A 598 A full-length gE mutant carrying the mutation motif, Y569K and Y582A mutations, reduced localization to the trans-Golgi network.

[0512] YK-VZV-010 (SEQ ID NO: 31) encodes the full-length gE polypeptide, and its A 593 E 594 A 595 A 596 D 597 A 598 (SEQ ID NO: 183) The sequence is "S 593 E 594 S 595 T 596 D 597 T 598" motif (SEQ ID NO: 182). YK-VZV-010 replaced the Ser / Thr-rich "SSTT" acidic cluster with an Ala-rich sequence; in addition, the YK-VZV-010 mRNA vaccine also had the Y569K and Y582A mutations. The results of Example 8 demonstrated that the cell membrane expression of the gE polypeptide of YK-VZV-010 was enhanced.

[0513] Each of the above mutants had modifications that reduced localization of the encoded gE protein to the trans-Golgi network and enhanced transport to the serosal membrane.

[0514] [Table 36]

[0515] Summary: In summary, the present invention designed 43 antigen sequences encoding VZV gE protein variants and screened several sequences with superior efficacy, including several antigen sequences that were more effective than at least the vaccines Shingrix® and YK-VZV-004 (Comparative Example 1), YK-VZV-006 (Comparative Example 2), YK-VZV-045 (Comparative Example 3), and YK-VZV-007 (Comparative Example 4). These results were surprising. Among these sequences, the sequences with the following mutation combinations were more preferred: YK-VZV-010 (full-length gE, Y569K, Y582A, S593A, S595A, T596A, T598A), YK-VZV-011 (full-length gE, A568D, Y569K, R570E, V571K), YK-VZV-013 (full-length gE, A568D, Y569K, R570E, V571K, Y582A), YK-VZV-018 (gE1-573aa, A568D), and YK-VZV-020 (gE1-573aa, Y569K). The table below shows the complete sequence information and its sequence number of this application. Taking the antigen sequence of YK-VZV-001 as an example, SEQ ID NO: 2 and SEQ ID NO: 3 are the nucleic acid sequence ("ORF-NT") and amino acid sequence ("ORF-AA") of the open reading frame of antigen YK-VZV-001, respectively, and SEQ ID NO: 1 and SEQ ID NO: 4 are the DNA sequence ("DNA") and mRNA sequence ("mRNA") of antigen YK-VZV-001, respectively.

[0516] [Table 37] TIFF2025179041000075.tif226170TIFF2025179041000076.tif226170TIFF2025179041000077.tif229170TIFF20251790410 00078.tif228170TIFF2025179041000079.tif228170TIFF2025179041000080.tif227170TIFF2025179041000081.tif100170

[0517] SEQ ID NO: 1 YK-VZV-001-DNA TCGACCTGCTGCTGGAATGGCTGTACGTGCCCATCGATCCTACCTGCCAGCCTATGCGGCTGTACTCCACCTGTCTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCTACAGAGTGGACAAGAGCCCTTACAACCAGAGCATGTACTACGCCGGCCTGCCTGTGGACGACTTCGAGGATAGCGAGAGCACCGACACCGAGGAAGAGTTCGGCAACGCCATTGGAGGATCTCACGGCGGCAGCAGCTATACCGTGTACATCGACAAGACCCGGTGAgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0518] sequence number 2 YK-VZV-001-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCA CAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCAGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGT ACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTG GTGGTGGATGTGGACTGCGCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGTTCGACGAGCTGGAACTGGACCCTCCTGAGATTGAACCCGGGGTGCTGAAGGTGCTGAGAACCGAGAAGCAGTACCTGGGAGTGTACATCTGGAACATGAGAGGCAGCGA CGGCACCTCTACCTACGCCACCTTTCTGGTCACATGGAAGGGCGACGAGAAAACACGGAACCCCACACCAGCTGTGACCCCTCAACCTAGAGGCGCCGAGTTTCACATGTGGAATTACCACAGCCACGTGTTCAGCGTGGGCGATACCTTTAGCCTGGCCATGCATCTGCAGTACAAGATCCACGAGGCCCCTTTCGACCTGCTGCTGGAATGGCTGTACGTGCCCATCGATCCTACCTGCCAGCCTATGCGGCTGTACTCCACCTGTCTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCTACAGAGTGGACAAGAGCCCTTACAACCAGAGCATGTACTACGCCGGCCTGCCTGTGGACGACTTCGAGGATAGCGAGAGCACCGACACCGAGGAAGAGTTCGGCAACGCCATTGGAGGATCTCACGGCGGCAGCAGCTATACCGTGTACATCGACAAGACCCGGTGA

[0519] Accession No. 3 YK-VZV-001-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPVDDFEDSESTDTEEEFGNAIGGSHGGSSYTVYIDKTR

[0520] Accession No. 4 YK-VZV-001-mRNA AGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGU

[0521] SEQ ID NO: 5 YK-VZV-002-DNA

[0522] SEQ ID NO: 6 YK-VZV-002-ORF-NT

[0523] sequence number 7 YK-VZV-002-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNG DDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPE IEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHM NSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYA

[0524] sequence number 8 YK-VZV-002 mRNA AAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUCAGAUAUGCCUGAGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0525] SEQ ID NO: 9 YK-VZV-003-DNA AATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAAGCCTGAgctggagcctcggtggccatgcttcttgccccttgggcctcc ccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0526] SEQ ID NO: 10 YK-VZV-003-ORF-NT AAGCCTAAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAAGCCTGA

[0527] sequence number 11 YK-VZV-003-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDD RHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPG VLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVAS TVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCVIFLICTAKRMRVKA

[0528] SEQ ID NO:12 YK-VZV-003 mRNA GGCGCCGAGUUUCACAUGUGGAAUUACCACAGCCACGUGUUCAGCGUGGGCGAUACCUUUAGCCUGGCCAUGCAUCUGCAGUACAAGAUCCACGAGGCCCCUUUCGACCUGCUGCUGGAAUGGCUGUACGUGCCCAUCGAUCCUACCUGCCAGCCUAUGCGGCUGUACUCCACCUGUCUGUAUCACCCCAACGCUCCCCAGUGCCUGAGCCACAUGAAUAGCGGCUGCACCUUCACAAGCCCUCACCUGGCUCAGCGAGUGGCCAGCACAGUGUACCAGAAUUGCGAGCACGCCGACAAUUACACCGCCUACUGUCUGGGCAUCAGCCACAUGGAACCUAGCUUCGGCCUGAUCCUGCACGAUGGCGGCACAACCCUGAAGUUCGUGGAUACCCCUGAGAGCCUGAGCGGCCUGUAUGUGUUCGUGGUGUACUUCAACGGCCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGAGAGUGAAAGCCUGAgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0529] SEQ ID NO: 13 YK-VZV-004-DNA AGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTGGTGGTGGATGTGGACTGCGCCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGTTCGACGAGCTGGAACTGGACCCTCCTGAGATTGAACCCGGGGTGCTGAAGGTGCTGAGAACCGAGAAGCAGTACCTGGGAGTGTACATCTGGAACATGAGAGGCAGCGACGGCACCTCTACCTACGCCACCTTTCTGGTCACA。 TGGAAGGGCGACGAGAAAACACGGAACCCCACACCAGCTGTGACCCCTCAACCTAGAGGCGCCGAGTTTCACATGTGGAATTACCACAGCCACGTGTTCAGCGTGGGCGATACCTTTAGCCTGGCCATGCATCTGCAGTACAAGATCCACGAGGCCCCTTTCGACCTGCTGCTGGAATGGCTGTACGTGCCCATCGATCCTACCTGCCAGCCTATGCGGCTGTACTCCACCTGTCTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAAGCCGCCAGAGTGGACAAGTGAgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0530] sequence number 14 YK-VZV-004-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCA CAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCAGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGT ACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTG GTGGTGGATGTGGACTGCGCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGTTCGACGAGCTGGAACTGGACCCTCCTGAGATTGAACCCGGGGTGCTGAAGGTGCTGAGAACCGAGAAGCAGTACCTGGGAGTGTACATCTGGAACATGAGAGGCAGCGA CGGCACCTCTACCTACGCCACCTTTCTGGTCCACATGGAAGGGCGACGAGAAAACCGGAACCCCACACCAGCTGTGACCCCTCAACCTAGAGGCGCGAGTTTCACATGTGGAATTACCACAGCCACGTGTTCAGCGTGGGCGATACCTTTAGCCTGGCCATGCATCTGCAGTACAAGATCCACGAGGCCCTTTCGACC TGCTGCTGGAATGGCTGTACGTGCCCATCGATCCTACCTGCCAGCCTATGCGGCTGTACTCCACCTGTCTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCT ACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGGCGGCCTGTATGTGTTCGTGGTGTACTTACAACGGCCACGTGGAAGCCGTCTACACCGTGGTGTCTACCGTGGACACTTCGTGAACGCCATCGAGGAAAGA GGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAAGCCGCCAGAGTGGGACAAGTGA

[0531] sequence no. 15 YK-VZV-004-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAARVDK

[0532] SEQ ID NO: 16 YK-VZV-004-mRNA AGAAAUAGAGAGAAAAGAGUAAGUAAGAAAUAUAAAGACCACCAUGGGCACCGUGAACAAGCCUUGUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAUUCCUGUGCGGGCUAGCGUGCUGAGAAUACGACGACGACUUCCACAUCGACGAGGACAAGCGUGUACGAGCCCUACUACCACAGCGUCGAUAGCGAUCGCCGAGUCUACUAGCCUAGCUGGCAACACAGAGGCGAGGAGACAGAAGGCCUACGAACCACAAGGCCUACCACAACAGCCCCUACAUCUGGCCCCCGGAACGACUACGACGAUGGCUUCCUGGAAAAUUGCCCACGAGCACCACGGCGUGGUACAAUCAAGGCAGGCAUCGACACGCGGCGAGAGACUGAUGCAGCCUA

[0533] Array No. 17 YK-VZV-006-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCC

[0534] SEQ ID NO: 18 YK-VZV-006-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGG

[0535] sequence number 19 YK-VZV-006-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSP YIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPA IQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVF SVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNGSCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVE AVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITVPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAARVDKSPYNQSMYYAGLPVDDFEDAEAADAEEEFGNAIGGSHGGSSYTVYIDKTR

[0536] SEQ ID NO:20 YK-VZV-006 mRNA AUUGCGAGCACGCCGACAAUUACACCGCCUACUGUCUGGGCAUCAGCCACAUGGAACCUAGCUUCGGCCUGAUCCUGCACGAUGGCGGCACAACCCUGAAGUUCGUGGAUACCCCUGAGAGCCUGAGCGGCCUGUAUGUGUUCGUGGUGUACUUCAACGGCCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGAGAGUGAAGGCCGCUAGAGUGGACAAGAGCCCUUACAACCAGAGCAUGUACUACGCCGGCCUGCCUGUGGACGAUUUCGAGGAUGCUGAAGCCGCCGAUGCCGAGGAAGAGUUUGGCAACGCCAUUGGCGGAUCUCACGGCGGCAGCAGCUAUACCGUGUACAUCGACAAGACCCGGUGAgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0537] SEQ ID NO: 21 YK-VZV-007-DNA

[0538] sequence number 22 YK-VZV-007-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAG AGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGCTTCCTGGAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACG TGATCCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGG TCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTGGTGGATGTGGACTGCGCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACAC

[0539] sequence number 23 YK-VZV-007-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYG DVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGGTST YATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVD TPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRWDKSPYNQSMYAAGLPVDDFEDAEAADAEEEFGNAIGGSHGGSSYTVYIDKTR

[0540] SEQ ID NO:24 YK-VZV-007 mRNA AGACUCCUCCCAUCCUCUCCCCUGUCCCUGUCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUC AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0541] SEQ ID NO: 25 YK-VZV-009-DNA AGGATAGCGAGAGCACCGACACCGAGGAAGAGTTCGGCAACGCCATTGGAGGATCTCACGGCGGCAGCAGCTATACCGTGTACATCGACAAGACCCGGTGAgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0542] Sequence No. 26 YK-VZV-009-ORF-NT AAGCCTAAAAGAAATCACACCCGGAATCCCGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCAAGAGAGTGGACAAGAGCCCTTACAACCAGTCTATGTATGCCGCCGGACTGCCCGTGGACGATTTCGAGGATAGCGAGAGACACCGACACCGAGAGAGAGTTCGGCAACGCCATTGGAGGATCTCACGGCGGCAGCAGCTATACCGTGTACATCGACAAGACCCGGTGA

[0543] sequence number 27 YK-VZV-009-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAKRVDKSPYNQSMYAAGLPVDDFEDSESTDTEEEFGNAIGGSHGGSSYTVYIDKTR

[0544] SEQ ID NO: 28 YK-VZV-009-mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGU ACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCUUCCUGGAAAUGCCCACGAGCACCACGGCGUGUACAAUCAAGGCAGAGGCAUCGACAGCGGCGAGAGACUGAUGCAGCCUACACAGAUGAGCGCCCAAGA GGACCUGGGAGAUGAUACCGGCAUCCACGUGAUCCCCACACUGAACGGCGACGACAGACACAAGAUCGUGAACGUGGACCAGCGGCAGUACGGCGACGUGUUCAAGGGCGACCUGAAUCCUAAGCCUCAGGGCCAGCGCCUGAUCGAGGUGUCCGUGGAAGAGAAUCACCCCUUCACACUGAGAGCCCCUAUCCAGAGAAUCUACGGCGUGCGCUAUACCGAGACAUGGUCCUUUCUGCCCAGCCUGACAUGUACCGGGGGAUGCCGCUCCUGCCAUCCAGCACAUUUGCCUGAAGCACACCACCUGUUUCCAGGACGUGGUGGUGGAUGUGGACUGCGCCGAGAACACCAAAGAGGAUCAGCUGGCCGAGAUCAGCUACCGGUUCCAGGGAAAGAAAGAAGGGCCGACCAGCCUUGGAUCGUGGUCAACACCAGCACACUGU

[0545] Array number 29 YK-VZV-010-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAA

[0546] Accession No. 30 YK-VZV-010-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGG

[0547] sequence number 31 YK-VZV-010-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENH PFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAY TVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAKRVDKSPYNQSMYAAGLPVDDFEDAEAADAEEEFGNAIGGSHGGSSYTVYIDKTR

[0548] SEQ ID NO:32 YK-VZV-010 mRNA AAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGAGAGUGAAGGCCAAGAGAGUGGACAAGAGCCCUUACAACCAGUCUAUGUAUGCCGCCGGACUGCCCGUGGACGAUUUUGAGGAUGCUGAAGCCGCCGACGCCGAGGAAGAAUUUGGCAACGCCAUUGGCGGAAGCCACGGCGGCAGUAGCUAUACCGUGUACAUCGACAAGACCCGGUGAgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0549] SEQ ID NO: 33 YK-VZV-011-DNA ATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGCGCGTGAAGGACAAAGAGAAGGACAAGTCCCCTTACAACCAGAGCATGTACTACGCCGGCCTGCCTGTGGACGACTTCGAGGATAGCGAGAGCACCGACACCGAGGAAGAGTTCGGCAACGCCATTGGAGGATCTCACGGCGGCAGCAGCTATACCGTGTACATCGACAAGACCCGGTGAgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0550] SEQ ID NO: 34 YK-VZV-011-ORF-NT ATGCATCTGCAGTACAAGATCCACGAGGCCCTTTCGACCTGCTGCTGGAATGGCTGTACGTGCCCATCGATCCTACCTGCCAGCCTATGCGGCTGTACTCCACCTGTCTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGC ACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGGCGGCCTGTATGTGTTCGTGGTGTACTTCACGGCCACGTGGAAGCCGTGGCCTACACC GTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTTGCCTGGTCATCTTCCTGATC TGCACCGCCAAGCGGATGCGCGTGAAGGACAAAGAGAAGGACAAGTCCCCTTACAACCAGAGCATGTACTACGCCGGCCTGCCTGTGGACGACTTCGAGGATAGCGAGAGCACCGACACCGAGGAAGAGTTCGGCAACGCCATTGGAGGATCTCACGGCGGCAGCAGCTATACCGTGTACATCGACAAGACCCGGTGA

[0551] sequence number 35 YK-VZV-011-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLDGDTGIHVIPTLNGDDRHKIVNVDQRQYG DVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTST YATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVD TPESLSGLYVFFVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITVPNPGTSPLLRYAAAWTGGLAAVVLLCLVIFLICTAKRMRVKDKEKDKSPYNQSMYYAGLPVDDFEDSESTDTEEEFGNAIGGSHGGSSYTVYIDKTR

[0552] sequence number 36 YK-VZV-011-mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCUUCCUGGAAAAUGCCCACGAGCACCACGGCGUGUACAAUCAAGGCAGAGGCAUCGACAGCG

[0553] Array number 37 YK-VZV-012-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATC AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0554] SEQ ID NO: 38 YK-VZV-012-ORF-NT GCTATACCGTGTACATCGACAAGACCCGGTGA

[0555] sequence number 39 YK-VZV-012-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYG DVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGGTST YATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVD TPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITVPNGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKDKEKDKSPYNQSMYYAGLPVDDFEDAEAADAEEEFGNAIGGSHGGSSYTVYIDKTR

[0556] SEQ ID NO:40 YK-VZV-012 mRNA

[0557] Array number 41 YK-VZV-013-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTGGTGGTGGATGTGGACTGCGCCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGT

[0558] sequence number 42 YK-VZV-013-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGCTTCCTGGGAAAATGCCCACGAGCACCACGG GTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGG

[0559] sequence number 43 YK-VZV-013-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYG DVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGGTST YATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVD TPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITVPNGTPLLRYAAAWTGGLAAVVLLCLVIFLICTAKRMRVKDKEKDKSPYNQSMYAAGLPVDDFEDSESTDTEEEFGNAIGGSHGGSSYTVYIDKTR

[0560] SEQ ID NO:44 YK-VZV-013 mRNA UACAUCGACAAGACCCGGUGAgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcU gagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0561] SEQ ID NO: 45 YK-VZV-014-DNA AATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGCGCGTGAAGGACAAAGAGAAGGACAAGTCCCCTTACAACCAGTCTATGTATGCCGCCGGACTGCCCGTGGACGATTTTGAGGATGCTGAAGCCGCCGACGCCGAGGAAGAATTTGGCAACGCCATTGGCGGAAGCCACGGCGGCAGTAGCTATACCGTGTACATCGACAAGACCCGGTGAgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0562] SEQ ID NO: 46 YK-VZV-014-ORF-NT TGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGGCGGCCTGTATGTGTTCGTGGTGTACTTCCAACGGCCACGTGGAAGCGTGGCCTACACCGTGGTGTCTACCGTGGACACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGC CGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGCGCGTGAAGGACAAAGAGAAGGACAAGTCCCCTTACAACCAGTCTATGTATGCCGCCGGACTGCCCGTGGACGATTTTGAGGATGCTGAAGCCGCCGACGCCGAGGAAGATTGGCAACGCCATTGGCGGAAGCCACGGCGGCAGTAGCTATACCGTGTACATCGACAAGACCCGGTGA

[0563] sequence number 47 YK-VZV-014-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLDGDTGIHVIPTLNGDDRHKIVNVDQRQYG DVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTST YATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVD TPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITVPNPGTSPLLRYAAAWTGGLAAVVLLCLVIFLICTAKRMRVKDKEKDKSPYNQSMYAAGLPVDDFEDAEAADAEEEFGNAIGGSHGGSSYTVYIDKTR

[0564] sequence number 48 YK-VZV-014-mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAG AUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCU UCCUGGAAAAUGCCCACGAGCACCACGGCGUGUACAAUCAAGGCAGAGGCAUCGACAGCGGCGAGAGACUGAUGCAGCCUACACAGAUGAGCGCCCAAGAGGACCUGGGAGAUGAUACCGGCAUCCACGUGAUCCCCACACUGAACGGCGACGACAGACACAAGAUCGUGAACGUGGACCAGCGGCAGUACGGCGACGUGUUCAAGGGCGACCUGAAUCCUAAGCCUCAGGGCCAGCGCCUGAUCGAGGUGUCCGUGGAAGAGAAUCACCCCUUCACACUGAGAGAGCCCCUAUCCAGAGAAUCUACGGCGCGUGCGCUAUACCGAGACAUGGUCCUUUCUGCC

[0565] Array number 49 YK-VZV-015-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCG

[0566] sequence number 50 YK-VZV-015-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCT

[0567] sequence number 51 YK-VZV-015-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENT KEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVAS TVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAMYYAGLPVDDFEDSESTDTEEFGNAIGGSHGGSSYTVYIDKTR

[0568] SEQ ID NO:52 YK-VZV-015 mRNA CCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGAGAGUGAAGGCCAUGUACUAUGCCGGCCUGCCUGUGGACGACUUCGAGGAUAGCGAGAGCACCGACACCGAGGAAGAGUUCGGCAACGCCAUUGGAGGAUCUCACGGCGGCAGCAGCUAUACCGUGUACAUCGACAAGACCCGGUGAgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0569] SEQ ID NO: 53 YK-VZV-016-DNA GTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCATGTACTATGCCGGCCTGCCTGTGGACGATTTCGAAGATGCTGAAGCCGCCGACGCCGAGGAAGAGTTTGGAAACGCCATTGGCGGAAGCCACGGCGGCAGCAGCTACACTGTGTACATCGACAAGACCCGGTGAgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0570] SEQ ID NO: 54 YK-VZV-016-ORF-NT CAGCCACGTGTTCAGCGTGGGCGATACCTTTAGCCTGGCCATGCATCTGCAGTACAAGATCCACGAGGCCCTTTCGACCTGCTGCTGGAATGGCTGTACGTGCCATCGATCCTACCTGCCAGCCTATGCGGCTGTACTCCACCTGTCTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGC ACCTTCACAAGCCCTCCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAAATTACACCGCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGGCGGCCTGTATGTGTTCGTGGT GTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGGAATCCCGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTG CTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCATGTACTATGCCGGCCTGCCTGTGGACGATTTCGAAGATGCTGAAGCCGCCGACGCCGAGGAAGAGTTTGGAAACGCCATTGGCGGAAGCCACGGCGGCAGCAGCTACACTGTGTACATCGACAAGACCCGGTGA

[0571] sequence number 55 YK-VZV-016-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLDGDTGIHVIPTLNGDDRHKIVNVDQRQ YGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGS DGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDG GTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITVPNPGTSPLLRYAAAWTGGLAAVVLLCLVIFLICTAKRMRVKAMYYAGLVPDDFEDAEAADAEEEFGNAIGGSHGGSSYTVYIDKTR

[0572] sequence number 56 YK-VZV-016-mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCUUCCUGGAAAAUGCCCACGAGCACCACGGCGUGUACAAUCAAGGCAGAGGCAUCGACAGCG

[0573] Array number 57 YK-VZV-017-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTA

[0574] SEQ ID NO: 58 YK-VZV-017-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGG

[0575] sequence number 59 YK-VZV-017-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLDGDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETW SFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPT CQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITVPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDK

[0576] SEQ ID NO:60 YK-VZV-017 mRNA CCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGAGAGUGAAGGCCUACAGAGUGGACAAGUGAUAAUAGgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0577] SEQ ID NO: 61 YK-VZV-018-DNA CATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGACTACAGAGTGGACAAGTGATAATAGgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0578] Accession No. 62 YK-VZV-018-ORF-NT AGTGTACCCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGGCGGCCTGTATGTGTTCGTGGTGTACTTCCAACGGCCACGTGGAAGCGTGGCCTACACCGTGGTGTCTACCGTGGACACTTCG TGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGACTACAGAGTGGACAAGTGA

[0579] sequence number 63 YK-VZV-018-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVL KVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVY QNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKDYRVDK

[0580] SEQ ID NO:64 YK-VZV-018 mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCG ACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCUUCCUGGAAAAUGCCCACGAGCACCACGGCGUGUACAAUCA AGGCAGAGGCAUCGACAGCGGCGAGAGACUGAUGCAGCCUACACAGAUGAGCGCCCAAGAGGACCUGGGAGAUGAUACCGGCAUCCACGUGAUCCCCACACUGAACGGCGACGACAGACACAAGAUCGUGAACGUGGACCAGCGGCAGUACGGCGACGUGUUCAAGGGCGACCUGAAUCCUAAGCCUCAGGGCCAGCGCCUGAUCGAGGUGUCCGUGGAAGAGAAUCACCCCUUCACACUGAGAGCCCCUAUCCAGAGAAUCUACGGCGUGCGCUAUACCGAGACAUGGUCCUUUCUGCCCAGCCUGACAUGUACCGGGGGAUGCCGCUCCUGCCAUCCAGCACAUUUGCCUGAAGCACACCACCUGUUUCCAGGACGUGG

[0581] Accession No. 65 YK-VZV-019-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAA

[0582] Accession number 66 YK-VZV-019-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGG

[0583] sequence number 67 YK-VZV-019-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVL KVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVY QNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKYRVDK

[0584] SEQ ID NO:68 YK-VZV-019 mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUC

[0585] SEQ ID NO: 69 YK-VZV-020-DNA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0586] SEQ ID NO: 70 YK-VZV-020-ORF-NT

[0587] sequence number 71 YK-VZV-020-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVL KVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVY QNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAKRVDK

[0588] SEQ ID NO:72 YK-VZV-020 mRNA GUGCCUGAGCCACAUGAAUAGCGGCUGCACCUUCACAAGCCCUCACCUGGCUCAGCGAGUGGCCAGCACAGUGUACCAGAAUUGCGAGCACGCCGACAAUUACACCGCCUACUGUCUGGGCAUCAGCCACAUGGAACCUAGCUUCGGCCUGAUCCUGCACGAUGGCGGCACAACCCUGAAGUUCGUGGAUACCCCUGAGAGCCUGAGCGGCCUGUAUGUGUUCGUGGUGUACUUCAACGGCCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGAGAGUGAAGGCCAAGAGAGUGGACAAGUGAUAAUAGgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0589] SEQ ID NO: 73 YK-VZV-021-DNA GTACAAGATCCACGAGGCCCCTTTCGACCTGCTGCTGGAATGGCTGTACGTGCCCATCGATCCTACCTGCCAGCCTATGCGGCTGTACTCCACCTGTCTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCTATGAGGTGGACAAGTGATAATAGgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0590] Sequence number 74 YK-VZV-021-ORF-NT CAGCCACGTGTTCAGCGTGGGCGATACCTTTAGCCTGGCCATGCATCTGCAGTACAAGATCCACGAGGCCCTTTCGACCTGCTGCTGGAATGGCTGTACGTGCCATCGATCCTACCTGCCAGCCTATGCGGCTGTACTCACCTGTCTGTATCACCCCAACGCTCCCC AGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCA CAACCCTGAAGTTCGTGGATACCCCTGGAGAGCCTGAGCGGCCTGTATGGTTCGTGGTGACTTCCAACGGCCACGTGGAAGCCGTCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCT GCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCTATGAGGTGGACAAGTGA

[0591] sequence number 75 YK-VZV-021-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVL KVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVY QNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYEVDK

[0592] SEQ ID NO:76 YK-VZV-021 mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCUUCCUGGAAAAUGCCCACGAGCACCACGGCGUGUACAAUCAAGGCAGAGGCAUCGACAGCGGCGAGAGACUGAUGCAGCCUACACAGAUGAGCGCCCAAGAGGACCUGGGAGAUGAUACCGGCAUCCACGUGAUCCCCACACUGAACGGCGACGACAGACACAAGAUCGUGAACGUGGACC

[0593] Accession No. 77 YK-VZV-022-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCG

[0594] Accession number 78 YK-VZV-022-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAA

[0595] sequence number 79 YK-VZV-022-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLDGDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHIC LKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNGSCTFTSPHLAVASTVYQNCEHADNYTA YCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRKDK

[0596] SEQ ID NO:80 YK-VZV-022 mRNA GAGAGUGAAGGCCUACCGGAAGGACAAGUGAUAAAUAGgcUggagccUcggUggccaUgcUUcUUgccccUUugggccUccccccagccccUccUccUccUUccUgcacccgUaccccgUggUcUUUgaaUaaagUcUgagUgggcggAA

[0597] sequence number81 YK-VZV-023-DNA AATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGCGCGTGAAGGACAAAGAGAAGGACAAGTGATAATAGgctggagcctcggtggccatgcttcttgccc cttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0598] SEQ ID NO: 82 YK-VZV-023-ORF-NT AGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGCGCGTGAAGGACAAAGAGAAGGACAAGTGA

[0599] sequence number 83 YK-VZV-023-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVL KVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVY QNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKDKEKDK

[0600] SEQ ID NO:84 YK-VZV-023 mRNA 。

[0601] Array number 85 YK-VZV-024-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTGGTGGTGGATGTGGACTGCGCCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGT

[0602] sequence number 86 YK-VZV-024-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCT GGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCAGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCCCCAAGAGGACCTGGG AGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCCGCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGA ATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTGGTGGTGGATGGACTGCGCGCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGG GAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGTTCGACGAGCTGGAACTGGACCCTCCTGAGATTGAACCCGGGGTGCTGAAGGTGCTGAGAACCGAGAAGCAGTACCTGGGAGTGTACATCTGGAACATGAGAGGCAGCGACGGCACCTCTACCTACGCCACCTTTCTGGTCACATGGAAGGGCGACGAGAAAACACGGAACCCCACACCAGCTGTGACCCCTCAACCTAGAGGCGCCGAGTTTCACATGTGGAATTACCACAGCCACGTGTTCAGCGTGGGCGATACCTTTAGCCTGGCCATGCATCTGCAGTACAAGATCCACGAGGCCCCTTTCGACCTGCTGCTGGAATGGCTGTACGTGCCCATCGATCCTACCTGCCAGCCTATGCGGCTGTACTCCACCTGTCTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGACGCCGAGAAGGACAAGTGA

[0603] Accession number 87 YK-VZV-024-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKDAEKDK

[0604] Accession number 88 YK-VZV-024-mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGC

[0605] Array number 89 YK-VZV-025-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATC

[0606] SEQ ID NO: 90 YK-VZV-025-ORF-NT

[0607] sequence number 91 YK-VZV-025-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLDGDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETW SFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPT CQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKKEKDK

[0608] SEQ ID NO:92 YK-VZV-025 mRNA CCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGCGCGUGAAGAAAGAGAAGGACAAGUGAUAAUAGgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0609] ​​​ CATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCTACAGAGTGGACAAGAGCCCTTACAACCAGAGCATGTACTACGCCGGCCTGCCTGTTTGATAATAGgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0610] Sequence number 94 YK-VZV-026-ORF-NT CCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGGCGGCCTGTATGTGTTCGTGGTACTTTCAACGGCCACGTGGAAGCGTGGCCTACACCGTGGTGTCTACCGTGGACACTTCGT GAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCTACAGAGTGGACAAGAGCCCTTACAACCAGAGCATGTACTACGCCGGCCTGCCTGTTGA

[0611] sequence number 95 YK-VZV-026-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPV

[0612] SEQ ID NO: 96 YK-VZV-026-mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAG AUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCU UCCUGGAAAAUGCCCACGAGCACCACGGCGUGUACAAUCAAGGCAGAGGCAUCGACAGCGGCGAGAGACUGAUGCAGCCUACACAGAUGAGCGCCCAAGAGGACCUGGGAGAUGAUACCGGCAUCCACGUGAUCCCCACACUGAACGGCGACGACAGACACAAGAUCGUGAACGUGGACCAGCGGCAGUACGGCGACGUGUUCAAGGGCGACCUGAAUCCUAAGCCUCAGGGCCAGCGCCUGAUCGAGGUGUCCGUGGAAGAGAAUCACCCCUUCACACUGAGAGAGCCCCUAUCCAGAGAAUCUACGGCGCGUGCGCUAUACCGAGACAUGGUCCUUUCUGCC

[0613] Array No. 97 YK-VZV-027-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACC

[0614] Accession number 98 YK-VZV-027-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACG

[0615] sequence number 99 YK-VZV-027-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNG DDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPE IEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHL AQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICT AKRMRVKAMYYAGLPV

[0616] SEQ ID NO:100 YK-VZV-027 mRNA cccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0617] SEQ ID NO: 101 YK-VZV-028-DNA GAGAGTGAAGGCTATGTATGCCGCCGGACTGCCTGTTTGATAATAGgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctt tgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0618] SEQ ID NO: 102 YK-VZV-028-ORF-NT TGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCTATGTATGCCGCCGGACTGCCTGTTTGA

[0619] sequence number 103 YK-VZV-028-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRH KIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKV LRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAMYAAGLPV

[0620] SEQ ID NO:104 YK-VZV-028 mRNA UCAGCGUGGGCGAUACCUUUAGCCUGGCCAUGCAUCUGCAGUACAAGAUCCACGAGGCCCCUUUCGACCUGCUGCUGGAAUGGCUGUACGUGCCCAUCGAUCCUACCUGCCAGCCUAUGCGGCUGUACUCCACCUGUCUGUAUCACCCCAACGCUCCCCAGUGCCUGAGCCACAUGAAUAGCGGCUGCACCUUCACAAGCCCUCACCUGGCUCAGCGAGUGGCCAGCACAGUGUACCAGAAUUGCGAGCACGCCGACAAUUACACCGCCUACUGUCUGGGCAUCAGCCACAUGGAACCUAGCUUCGGCCUGAUCCUGCACGAUGGCGGCACAACCCUGAAGUUCGUGGAUACCCCUGAGAGCCUGAGCGGCCUGUAUGUGUUCGUGGUGUACUUCAACGGCCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGAGAGUGAAGGCUAUGUAUGCCGCCGGACUGCCUGUUUGAUAAUAGgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0621] SEQ ID NO: 105 YK-VZV-029-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTGGTGGTGGATGTGGACTGCGCCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGTTCGACGAGCTGGAACTGGACCCTCCTGAGATTGAACCCGGGGTGCTGAAGGTGCTGAGAACCGAGAAGCAGTACCTGGGAGTGTACATCTGGAACATGAGAGGCAGCGACGGCACCTCTA。

[0622] sequence number 106 YK-VZV-029-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGG CCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCAGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCACACTGAACGGCGACGACAGACACAAGATCGT GAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGATGCCGCCTGCCATCCAGCACATTTGC CTGAAGCACACCACCTGTTTCCAGGACGTGGTGGTGGATGTGGACTGCGCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGTTCGACGAGCTGGAACTGGACCCTCCTGAGATTGAACCCGGGGTGCTGAAGGTGCTGGAACCGAGAAGC AGTACCTGGGAGTGTACATCTGGAACATGAGAGGCAGCGACGGCACCTCTACCTACGCCACCTTTCTGGTCACATGGAAGGGCGACGAGAAAACAGGAACCCCACACCAGCTGTGACCCCTCAACCTAGAGGCGCGAGTTTCACATGTGGAATTACCACAGCCACGTGTTCAGCGTGGGCGATACCTTTAGCCTGGCCATGCATCTGCAG TACAAGATCCACGAGGCCCTTTCGACCTGCTGCTGGAATGGCTGTACGTGCCCATCGATCCTACCTGCCAGCCTATGCGGCGTACTCCACCTGTCTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCC GACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGGCGGCCTGTATGTGTTCGTGGTGTACTTACAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACACTTCGTGAACGCCATCGAGGAA AGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCTATGTATGGCCCGGACTGCCTGTTTTGA

[0623] sequence number 107 YK-VZV-029-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRH KIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKV LRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAMYGAGLPV

[0624] SEQ ID NO:108 YK-VZV-029 mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCUUCCUGGAAAAUGCCCACGAG

[0625] Array number 109 YK-VZV-030-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGC

[0626] Accession number 110 YK-VZV-030-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCC

[0627] sequence number 111 YK-VZV-030-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLDGDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETW SFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLY STCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKDKKDKSPYNQSMYAAGLPV

[0628] SEQ ID NO:112 YK-VZV-030 mRNA CCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGCGCGUGAAGGACAAAGAGAAGGACAAGUCCCCUUACAACCAGUCUAUGUAUGCCGCCGGACUGCCCGUUUGAUAAUAGgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0629] SEQ ID NO: 113 YK-VZV-031-DNA ATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGCGCGTGAAGGACAAAGAGAAGGACAAGTCCCCTTACAACCAGTCTATGTATGGCGCCGGACTGCCCGTTTGATAATAGgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0630] SEQ ID NO: 114 YK-VZV-031-ORF-NT AACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCTCTGTATGTGTTCGTGTGTACTTCCAACGGCCACGTGGAAGCGTGGCCTACACCGTGGTG TCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGCGCGTGAAGGACAAAGAGAAGGACAAGTCCCCTTACAACCAGTCTATGTATGGCCCGGACTGCCCGTTTGA

[0631] sequence number 115 YK-VZV-031-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKI VNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEK QYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTA YCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKDKEKDKSPYNQSMYGAGLPV

[0632] SEQ ID NO:116 YK-VZV-031 mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUA GCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCC CCGGAACGACUACGAUGGCUUCCUGGAAAAUGCCCACGAGCACCACGGCGUGUACAAUCAAGGCAGAGGCAUCGACAGCGGCGAGAGACUGAUGCAGCCUACACAGAUGAGCGCCCAAGAGGACCUGGGAGAUGAUACCGGCAUCCACGUGAUCCCCACACUGAACGGCGACGACAGACACAAGAUCGUGAACGUGGACCAGCGGCAGUACGGCGACGUGUUCAAGGGCGACCUGAAUCCUAAGCCUCAGGGCCAGCGCCUGAUCGAGGUGUCCGUGGAAGAGAAUCACCCCUUCACACUGAGAGAGCCCCUAUCCAGAGAA

[0633] Array No. 117 YK-VZV-032-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACA

[0634] Accession number 118 YK-VZV-032-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGG

[0635] sequence number 119 YK-VZV-032-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLDGDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHIC LKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNGSCTFTSPHLAVASTVYQNCEHADNYTA YCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKDYRVDKSPYNQSMYAAGLPV

[0636] SEQ ID NO:120 YK-VZV-032 mRNA GAGAGUGAAGGACUACAGAGUGGACAAGAGCCCUUACAACCAGUCUAUGUAUGCCGCGGACUGCCCGUUUGAUAAUAGgcUggagccUcggUggccaUgcUUcUUgccccUugggccUccccccagccccUccUccccUUccUUccUgcacccgUaccccgUggUcUUUgaaUaaagUcUgagUgggcggcgAA

[0637] sequence number 121 YK-VZV-033-DNA CAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCAAGAGAGTGGACAAGAGCCCTTACAACCAGTCTATGTATGCCGCCGGACTGCCCGT TTGATAATAGgctggagcctcggtggccatgcttcttgccccttgggcctcccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0638] sequence number 122 YK-VZV-033-ORF-NT TACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCAAGAGAGTGGACAAGAGCCCTTACAACCAGTCTATGTATGCCGCCGGACTGCCGTTTGA

[0639] sequence number 123 YK-VZV-033-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKI VNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEK QYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTA YCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITVPNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAKRVDKSPYNQSMYAAGLPV

[0640] Accession number 124 YK-VZV-033-mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCUUCCUGGAAAAUGCCCACGAGCACCACGGCGUGUACAAUCAAGGCAGAGGCAUCGACAGCGGCGAGAGACUGAUGCAGCCUACACAGAUGAGCGCCCAAGAGGACCUGGGAGAUGAUACCGGCAUCCACGUGAUCCCCACACUGAACGGCGACGACAGACACAAGAUCGUGAACGUGGACCAGCGGCAGUACGGCGACGUGUUCAAGGGCGACCUGAAUCCUAAGCCUCAGGGCCAGCGCCUGAUCGAGGUGUCCGUGGAAGAGAAUCACCCCUUCACACUGAGAGCCCCUAUCCAGAGAAUCUACGGCGUGCGCUAUACCGAGACAUGGUCCUUUCUGCCCAGCCUGACAUGUACCGGGGAUGCCGCUCCUGCCAUCCAGCACAUUUGCCUGAAGCACACCACCUGUUUCCAGGACGUGGUGGUGGAUGUGGACUGCGCCGAGAACACCAAAGAGGAUCAGCUGGCCGAGAUCAGCUACCGGUUCCAGGGAAAGAAAGAGGCCGACCAGCCUUGGAUCGUGGUCAACACCAGCACACUGU

[0641] Array No. 125 YK-VZV-034-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAG

[0642] Array number 126 YK-VZV-034-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCC

[0643] sequence number 127 YK-VZV-034-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKI VNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEK QYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTA YCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYEVDKSPYNQSMYAAGLPV

[0644] SEQ ID NO:128 YK-VZV-034 mRNA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0645] SEQ ID NO: 129 YK-VZV-035-DNA ctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0646] SEQ ID NO: 130 YK-VZV-035-ORF-NT GCCAAGCGGATGAGAGTGAAGGCCTACCGGAAGGACAAGAGCCCTTTACAACCAGTCTATGTATGCCGCCGGACTGCCCGTTTGA

[0647] sequence number 131 YK-VZV-035-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKI VNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEK QYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTA YCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRKDKSPYNQSMYAAGLPV

[0648] SEQ ID NO:132 YK-VZV-035 mRNA UCAGCGUGGGCGAUACCUUUAGCCUGGCCAUGCAUCUGCAGUACAAGAUCCACGAGGCCCCUUUCGACCUGCUGCUGGAAUGGCUGUACGUGCCCAUCGAUCCUACCUGCCAGCCUAUGCGGCUGUACUCCACCUGUCUGUAUCACCCCAACGCUCCCCAGUGCCUGAGCCACAUGAAUAGCGGCUGCACCUUCACAAGCCCUCACCUGGCUCAGCGAGUGGCCAGCACAGUGUACCAGAAUUGCGAGCACGCCGACAAUUACACCGCCUACUGUCUGGGCAUCAGCCACAUGGAACCUAGCUUCGGCCUGAUCCUGCACGAUGGCGGCACAACCCUGAAGUUCGUGGAUACCCCUGAGAGCCUGAGCGGCCUGUAUGUGUUCGUGGUGUACUUCAACGGCCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGAGAGUGAAGGCCUACCGGAAGGACAAGAGCCCUUACAACCAGUCUAUGUAUGCCGCCGGACUGCCCGUUUGAUAAUAGgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0649] Array No. 133 YK-VZV-036-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTGGTGGTGGATGTGGACTGCGCCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGTTCGACGAGCTGGAACTGGACCCTCCTGAGATTGAACCCGGGGTGCTGAAGGTGCTGAGAACCGAGAAGCAGTACCTGGGA

[0650] sequence number 134 YK-VZV-036-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCC GAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATG AGGCGCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCAC CCCTTCACACTGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTGGTGGTGGATGGACTGCGCCGAGAACACC

[0651] sequence number 135 YK-VZV-036-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLDGDTGIHVIPTLNGDDRHKIVNVDQRQ YGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGS DGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDG GTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAMYAAGLPVDDFEDSESTDTEEEFGNAIGGSHGGSSYTVYIDKTR

[0652] SEQ ID NO:136 YK-VZV-036 mRNA AGACUCCUCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGU AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0653] SEQ ID NO: 137 YK-VZV-037-DNA TTGGCAACGCCATTGGCGGAAGCCACGGCGGCAGTAGCTATACCGTGTACATCGACAAGACCCGGTGATAATAGgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgt accccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0654] SEQ ID NO: 138 YK-VZV-037-ORF-NT CACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCTATGTATGCCGCCGGACTGCCCGTGGACGATTTTGAGGATGCTGAAGCCGCCGACGCCGAGGAAGAATTGGCAACGCCATTGGCGGAAGCCACGGCGGCAGTAGCTATACCGTGTACATCGACAAGACCCGGTGA

[0655] sequence number 139 YK-VZV-037-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQ YGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPVIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGS DGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDG GTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAMYAAGLPVDDFEDAEAADAEEEFGNAIGGSHGGSSYTVYIDKTR

[0656] Array No. 140 YK-VZV-037-mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCUUCCUGGAAAAUGCCCACGAGCACCACGGCGUGUACAAUCAAGGCAGAGGCAUCGACAGCGGCGAGAGACUGAUGCAGCCUACACAGAUGAGCGCCCAAGAGGACCUGGGAGAUGAUACCGGCAUCCACGUGAUCCCCACACUGAACGGCGACGACAGACACAAGAUCGUGAACGUGGACCAGCGGCAGUACGGCGACGUGUUCAAGGGCGACCUGAAUCCUAAGCCUCAGGGCCAGCGCCUGAUCGAGGUGUCCGUGGAAGAGAAUCACCCCUUCACACUGAGAGCCCCUAUCCAGAGAAUCUACGGCGUGCGCUAUACCGAGACAUGGUCCUUUCUGCCCAGCCUGACAUGUACCGGGGAUGCCGCUCCUGCCAUCCAGCACAUUUGCCUGAAGCACACCACCUGUUUCCAGGACGUGGUGGUGGAUGUGGACUGCGCCGAGAACACCAAAGAGGAUCAGCUGGCCGAGAUCAGCUACCGGUUCCAGGGAAAGAAAGAGGCCGACCAGCCUUGGAUCGUGGUCAACACCAGCACACUGUUCGACGAGCUGGAACUGGACCCUCCUGAGAUUGAACCCGGGGUGCUGAAGGUGCUGAGAACCGAGAAGCAGUACCUGGGA

[0657] Accession No. 141 YK-VZV-038-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAG

[0658] Accession number 142 YK-VZV-038-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGT

[0659] sequence number 143 YK-VZV-038-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENT KEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVV YFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKDKEKDKSPYNQSMYGAGLPVDDFEDSESTDTEEEFGNAIGGSHGGSSYTVYIDKTR

[0660] SEQ ID NO:144 YK-VZV-038 mRNA GCGCGUGAAGGACAAAGAGAAGGACAAGUCCCCUUACAACCAGUCUAUGUAUGGCGCCGGACUGCCCGUGGGACGAUUUCGAGGAUAGCGAGAGCACCGACACCGAGGAAGAGUUCGGCAACGCCAUUGGAGGAUCUCACGGCGGCAGCAGCUAUACCGUGUACAUCGACAAGACCCGGUGAUAAUAGgcUggagccUcggUgg ccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0661] SEQ ID NO: 145 YK-VZV-039-DNA GCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGCGCGTGAAGGACAAAGAGAAGGACAAGTCCCCTTACAACCAGTCTATGTATGGCGCCGGACTGCCCGTGGACGATTTTGAGGATGCTGAAGCCGCCGACGCCGAGGAAGAATTTGGCAACGCCATTGGCGGAAGCCACGGCGGCAGTAGCTATACCGTGTACATCGACAAGACCCGGTGATAATAGgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0662] SEQ ID NO: 146 YK-VZV-039-ORF-NT AACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTG CACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGGCGGCCTGTATGTGTTCGTGGTGTACTTCACGGCCACGTGGAAGCGTGGCCTACACCGTGGTGTCTACCGTGGACACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACT GCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGCGCGTGAAGGAC AAAGAGAAGGACAAGTCCCCTTACAACCAGTCTATGTATGGCGCGGACTGCCCGTGGACGATTTTGAGGATGCTGAAGCCGCCGACGCCGAGGAAGAATTGGCAACGCCATTGGCGGAAGCCACGGCGGCAGTAGCTATACCGTGTACATCGACAAGACCCGGTGA

[0663] sequence number 147 YK-VZV-039-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLDGDTGIHVIPTLNGDDRHKIVNVDQRQYG DVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTST YATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVD TPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITVPNPGTSPLLRYAAAWTGGLAAVVLLCLVIFLICTAKRMRVKDKEKDKSPYNQSMYGAGLPVDDFEDAEAADAEEEFGNAIGGSHGGSSYTVYIDKTR

[0664] sequence number 148 YK-VZV-039-mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCUUCCUGGAAAAUGCCCACGAGCACCACGGCGUGUACAAUCAAGGCAGAGGCAUCGACAGCGGCGAGAGACUGAUGCAGCCUACACAGAUGAGCGCCCAAGAGGACCUGGGAGAUGAUACCGGCAUCCACGUGAUCCCCACACUGAACGGCGACGACAGACACAAGAUCGUGAACGUGGACC

[0665] Accession No. 149 YK-VZV-040-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGC

[0666] SEQ ID NO: 150 YK-VZV-040-ORF-NT

[0667] sequence number 151 YK-VZV-040-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYG DVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGGTST YATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVD TPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITVPNGTPLLRYAAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRWDKSPYNQSMYGAGLPVDDFEDAEAADAEEEFGNAIGGSHGGSSYTVYIDKTR

[0668] SEQ ID NO:152 YK-VZV-040 mRNA GUGCCUGAGCCACAUGAAUAGCGGCUGCACCUUCACAAGCCCUCACCUGGCUCAGCGAGUGGCCAGCACAGUGUACCAGAAUUGCGAGCACGCCGACAAUUACACCGCCUACUGUCUGGGCAUCAGCCACAUGGAACCUAGCUUCGGCCUGAUCCUGCACGAUGGCGGCACAACCCUGAAGUUCGUGGAUACCCCUGAGAGCCUGAGCGGCCUGUAUGUGUUCGUGGUGUACUUCAACGGCCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGAGAGUGAAGGCCUACAGAGUGGACAAGAGCCCUUACAACCAGUCUAUGUAUGGCGCCGGACUGCCCGUGGACGAUUUUGAGGAUGCUGAAGCCGCCGACGCCGAGGAAGAAUUUGGCAACGCCAUUGGCGGAAGCCACGGCGGCAGUAGCUAUACCGUGUACAUCGACAAGACCCGGUGAUAAUAGgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0669] SEQ ID NO: 153 YK-VZV-041-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTGGTGGTGGATGTGGACTGCGCCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGTTCGACGAGCTGGAACTGGACCCTCCTGAGATTGAACCCGGGGTGCTGAAGGTGCTGAGAACCGAGAAGCAGTACCTGGGAGTGTACATCTGGAACATGAGAGGCAGCGACGGCACCTCTA。

[0670] sequence number 154 YK-VZV-041-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGC AGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCACACTGAACGGC GACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGG GATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACCACCTGTTTCCAGGACGTGGTGGTGGATGGACTCGCCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGTTCGACGAGCTGGAACTGGACCCTCCTGAG ATTGAACCCGGGGTGCTGAAGGTGCTGAGAACCGAGAAGCAGTACCTGGGAGTGTACATCTGGAACATGAGAGGCAGCGACGGCACCTCTACCTACGCCACCTTTCTGGTCACATGGAAGGGCGACGAGAAAACAGGAACCCCACACCAGCTGTGACCCCTCAACCTAGAGGCGCGAGTTTCACATGTGGAATTACCAAGCCACGTGTTCAGCGTGGGCGATAC CTTTAGCCTGGCCATGCATCTGCAGTACAAGATCCACGAGGCCCTTTCGACCTGCTGCTGGAATGGCTGTACGTGCCATCGATCCTACCTGCCAGCTATGCGGCTGTACTCCACCTGTCTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATT GCGAGCACGCCGACAAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGGCGGCCTGTATGTGTTCGTGGTGTACTTCACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACACTTCGTGAACGCCATCGAGGAAAGA GGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCGACGATTTCGAGACAGCGAGAGCACCGATACCGAGGAAGAAT

[0671] sequence number 155 YK-VZV-041-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKADDFEDSESTDTEEE

[0672] SEQ ID NO: 156 YK-VZV-041-mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCC

[0673] Array number 157 YK-VZV-042-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTA

[0674] SEQ ID NO: 158 YK-VZV-042-ORF-NT

[0675] sequence number 159 YK-VZV-042-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLDGDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKG DLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAP FDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNGSCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKADDFEDAEAADAEEE

[0676] SEQ ID NO:160 YK-VZV-042 mRNA GCCUGAGCGGCCUGUAUGUGUUCGUGGUGUACUUCAACGGCCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGAGAGUGAAGGCCGACGAUUUCGAGGAUGCCGAGGCCGCCGAUGCUGAGGAAGAAUGAUAAUAGgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0677] SEQ ID NO: 161 YK-VZV-043-DNA CCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCGACGATTTCGAGGACAGCGAGAGCACCGATACCGAGGAAGAGTTCGGCAACGCCATTGGCGGATCTCACGGCGGCAGCAGCTATACCGTGTACATCGACAAGACCCGGTGATAATAGgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0678] SEQ ID NO: 162 YK-VZV-043-ORF-NT TGCTGCTGGAATGGCTGTACGTGCCCATCGATCCTACCTGCCAGCCTATGCGGCTGACTCCACCTGTCTGTTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAAT TGCGAGCACGCCGACAAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGGCGGCCTGTATGTGTTCGTGGTGTACTTCACGGCCACGTGGAAGCCGTG GCCTACACCGTGGTGTCTACCGTGGACACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCT GTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCGACGATTTCGAGGACAGCGAGAGCACCGATACCGAGGAAGAGTTCGGCAACGCCATTGGCGGATCTCACGGCGGCAGCAGCTATACCGTGTACATCGACAAGACCCGGTGA

[0679] sequence number 163 YK-VZV-043-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQ RQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWN MRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNGSCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFG LILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKADDFEDSESTDTEEEFGNAIGGSHGGSSYTVYIDKTR

[0680] SEQ ID NO:164 YK-VZV-043 mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCAGCAGAAAGGCCUACGACCACAACAGCCCCUACAUCUGGCCCCGGAACGACUACGAUGGCUUCCUGGAAAAUGCCCACGAGCACCACGGCGUGUACAAUCAAGGCAGAGGCAUCGACAGCGGCGAGAGACUGAUGCAGCCUACACAGAUGAGCGCCCAAGAGGACCUGGGAGAUGAUACCGGCAUCCACGUGAUCCCCACA

[0681] SEQ ID NO: 165 YK-VZV-044-DNA AGACTCCTCCCCATCCTCTCCCTCTGTCCCTCTGTCCCTCTGACCCTGCACTGTCCCAGCACCGCCACCATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCA

[0682] Accession No. 166 YK-VZV-044-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCC

[0683] Accession number 167 YK-VZV-044-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKG DLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKADDFEDAEAADAEEEFGNAIGGSHGGSSYTVYIDKTR

[0684] Accession number 168 YK-VZV-044-mRNA GAUGGCGGCACAACCCUGAAGUUCGUGGAUACCCCUGAGAGCCUGAGCGGCCUGUAUGUGUUCGUGGUGUACUUCAACGGCCACGUGGAAGCCGUGGCCUACACCGUGGUGUCUACCGUGGACCACUUCGUGAACGCCAUCGAGGAAAGAGGCUUCCCUCCAACUGCUGGACAGCCUCCUGCCACCACCAAGCCUAAAGAAAUCACACCCGUGAAUCCCGGCACAAGCCCACUGCUUAGAUACGCCGCUUGGACAGGCGGACUGGCUGCUGUUGUUCUGCUGUGCCUGGUCAUCUUCCUGAUCUGCACCGCCAAGCGGAUGAGAGUGAAGGCCGACGAUUUCGAGGAUGCCGAGGCCGCCGAUGCUGAGGAAGAAUUUGGCAACGCCAUUGGCGGCUCUCACGGCGGCAGUAGCUAUACCGUGUACAUCGACAAGACCCGGUGAUAAUAGgcUggagccUcggUggccaUgcUUcUUgccccUUgggccUccccccagccccUccUccccUUccUgcacccgUacccccgUggUcUUUgaaUaaagUcUgagUgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0685] SEQ ID NO: 169 YK-VZV-045-DNA GGCTGTACTCCACCTGTCTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTGAGCGGCCTGTATGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTGTCTACCGTGGACCACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTTGTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCTACAGAGTGGACAAGAGCCCTTACAACCAGAGCATGTACGCTGCCGGCCTGCCTGTGGACGACTTCGAGGATAGCGAGAGCACCGACACCGAGGAAGAGTTCGGCAACGCCATTGGAGGATCTCACGGCGGCAGCAGCTATACCGTGTACATCGACAAGACCCGGTGAgctggagcctcggtggccatgcttcttgccccttgggcctccccccagcccctcctccccttcctgcacccgtacccccgtggtctttgaataaagtctgagtgggcggcAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[0686] SEQ ID NO: 170 YK-VZV-045-ORF-NT ATGGGCACCGTGAACAAGCCTGTTGTGGGCGTGCTGATGGGCTTCGGCATCATCACAGGCACCCTGCGGATCACCAATCCTGTGCGGGCTAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAGCCCTACTACCACAGCGATCACGCCGAGTCTAGCTGGGTCAACAGAGGCGAGAGCAGCAGAAAGGCCTACGACCACAACAGCCCCTACATCTGGCCCCGGAACGACTACGATGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAATCAAGGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCTACACAGATGAGCGCCCAAGAGGACCTGGGAGATGATACCGGCATCCACGTGATCCCCACACTGAACGGCGACGACAGACACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGACGTGTTCAAGGGCGACCTGAATCCTAAGCCTCAGGGCCAGCGCCTGATCGAGGTGTCCGTGGAAGAGAATCACCCCTTCACACTGAGAGCCCCTATCCAGAGAATCTACGGCGTGCGCTATACCGAGACATGGTCCTTTCTGCCCAGCCTGACATGTACCGGGGATGCCGCTCCTGCCATCCAGCACATTTGCCTGAAGCACACCACCTGTTTCCAGGACGTGGTGGTGGATGTGGACTGCGCCGAGAACACCAAAGAGGATCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGAAAGAAAGAGGCCGACCAGCCTTGGATCGTGGTCAACACCAGCACACTGTTCGACGAGCTGGAACTGGACCCTCCTGAGATTGAACCCGGGGTGCTGAAGGTGCTGAGAACCGAGAAGCAGTACCTGGGAGTGTACATCTGGAACATGAGAGGCAGCGACGGCACCTCTACCTACGCCACCTTTCTGGTCACATGGAAG GGCGACGAGAAAACAGGAACCCCACACCAGCTGTGACCCCTCAACCTAGAGGCGCGAGTTTCACATGTGGAATTACCACAGCCACGTGTTCAGCGTGGGCGATACCTTTAGCCTGGCCATGCATCTGCAGTACAAGATCCACGAGGCCCTTTCGACCTGCTGCTGGAATGGCTGTACGTGCCATCGATCCTACCTGCCAGCCTAGCGGCTGTACTCCACCTGT CTGTATCACCCCAACGCTCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACAAGCCCTCACCTGGCTCAGCGAGTGGCCAGCACAGTGTACCAGAATTGCGAGCACGCCGACAATTACACCGCTACTGTCTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGATGGCGGCACAACCCTGAAGTTCGTGGATACCCCTGAGAGCCTG AGCGGCCTGTATGTGTTCGTGGTGTACTTCCAACGGCCACGTGGAAGCGTGGCCTACACCGTGGTGTCTACCGTGGACACTTCGTGAACGCCATCGAGGAAAGAGGCTTCCCTCCAACTGCTGGACAGGCCTCCTGCCACCACCAAGCCTAAAGAAATCACACCCGTGAATCCCGGCACAAGCCCACTGCTTAGATACGCCGCTTGGACAGGCGGACTGGCTGCTGTT GTTCTGCTGTGCCTGGTCATCTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCTACAGAGTGGACAAGAGCCCTTACAACCAGAGCATGTACGCTGCCGGCCTGCCTGTGGACGACTTCGAGGATAGCGAGAGCACCGACCACGAGGAAGAGTTCGGCAACGCCATTGGAGGATCTCACGGCGGCAGCAGCTATACCGTGTACATCGACAAGACCCGGTGA

[0687] sequence number 171 YK-VZV-045-ORF-AA MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYAAGLPVDDFEDSESTDTEEEFGNAIGGSHGGSSYTVYIDKTR

[0688] Sequence number 172 YK-VZV-045-mRNA AGACUCCUCCCCAUCCUCUCCCUCUGUCCCUCUGUCCCUCUGACCCUGCACUGUCCCAGCACCGCCACCAUGGGCACCGUGAACAAGCCUGUUGUGGGCGUGCUGAUGGGCUUCGGCAUCAUCACAGGCACCCUGCGGAUCACCAAUCCUGUGCGGGCUAGCGUGCUGAGAUACGACGACUUCCACAUCGACGAGGACAAGCUGGACACCAACAGCGUGUACGAGCCCUACUACCACAGCGAUCACGCCGAGUCUAGCUGGGUCAACAGAGGCGAGAGCA

[0689] SEQ ID NO: 173 5' UTR 5′AGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACC3′ ...

Claims

1. comprising varicella-zoster virus (VZV) ribonucleic acid (RNA), said RNA encoding wild-type VZV gE glycoprotein or a mutant thereof; wherein the sequence of the wild-type VZV gE glycoprotein is SEQ ID NO: 3; wherein the VZV gE glycoprotein mutant is selected from YK-VZV-020, YK-VZV-030, YK-VZV-031, YK-VZV-009, YK-VZV-010, YK-VZV-011, YK-VZV-012, YK-VZV-013, and YK-VZV-014. Table 1

2. the variant of VZV gE glycoprotein is selected from YK-VZV-020, YK-VZV-010, YK-VZV-011 and YK-VZV-013; Table 2 Or, the amino acid sequence of the variant of VZV gE glycoprotein is as set forth in SEQ ID NO: 27, 31, 35, 39, 43, 47, 71, 111 or 115; or the VZV RNA sequence is as set forth in SEQ ID NO: 28, 32, 36, 40, 44, 48, 72, 112 or 116; or The VZV RNA is mRNA, or 2. The composition of claim 1, wherein the RNA sequence encoding the VZV gE glycoprotein corresponds to the DNA sequence set forth in SEQ ID NO: 25, 29, 33, 37, 41, 45, 69, 109, or 113.

3. 2. The composition of claim 1, wherein the VZV RNA has an open reading frame (ORF) encoding the VZV gE glycoprotein, and the sequence of the open reading frame is set forth in SEQ ID NO: 26, 30, 34, 38, 42, 46, 70, 110, or 114.

4. The composition of claim 1 , wherein the VZV RNA further comprises a 5′ untranslated region (UTR).

5. The composition of claim 4, wherein the sequence of the 5'UTR is set forth in SEQ ID NO: 173, 174, 175, 176 or 177.

6. The composition of claim 1 , wherein the VZV RNA further comprises a 3′ untranslated region (UTR).

7. The composition of claim 6, wherein the sequence of the 3'UTR is set forth in SEQ ID NO: 178, 179, 180 or 181.

8. The composition of claim 1 , wherein the VZV RNA further comprises a poly(A) tail.

9. 9. The composition of claim 8, wherein the poly(A) tail has a length of 50 to 150 nucleotides.

10. The composition of claim 1 , wherein the VZV RNA further comprises a 5′-terminal cap.

11. The composition of claim 10, wherein the 5' end cap is 7mG(5')ppp(5')NlmpNp.

12. The composition of claim 3 , wherein the sequence of the open reading frame is codon-optimized.

13. The composition of claim 12, wherein the sequence of the open reading frame comprises at least one base modification.

14. 14. The composition of claim 13, wherein the base modifications are selected from one or more of pseudouridine, N1-methylpseudouridine, N1-ethylpseudouridine, 2-thiouridine, 4'-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methoxyuridine, and 2'-O-methyluridine.

15. 15. The composition of claim 14, wherein the base modification is a replacement of uracil with pseudouridine and / or N1-methylpseudouridine.

16. The composition of claim 13, 14 or 15, wherein the base modifications are 1 to 100% base modifications.

17. 17. The composition of claim 16, wherein the base modifications are 100% base modifications.

18. providing a template capable of being transcribed into RNA of said VZV; transcribing using said template under conditions suitable for transcription into said RNA. A method for preparing a composition according to any one of claims 1 to 17, comprising:

19. 20. The method of claim 18, further comprising a purification step selected from lithium chloride precipitation, affinity chromatography, solution exchange by ultrafiltration, and cellulose chromatography.

20. 10. The composition of claim 1, which is a vaccine and further comprises a pharmaceutically acceptable carrier.

21. 21. The composition of claim 20, wherein the carrier comprises a lipid mixture.

22. 22. The composition of claim 21, wherein the lipid mixture is a lipid nanoparticle (LNP).

23. 23. The composition of claim 20, 21 or 22, wherein the vaccine is an mRNA vaccine.

24. 24. The composition of claim 23, wherein the lipid nanoparticles comprise cationic lipids, neutral lipids, structured lipids, and polymer-conjugated lipids.

25. the cationic lipid is selected from YK-009, YK-401, YK-305, ALC0315, SM102 and DLIN-MC3-DM; 【Chemistry 1】 【change】 Or, the molar ratio of the cationic lipid to the neutral lipid is (1 to 10):1; Or, the molar ratio of the cationic lipid to the structural lipid is (1-5):1; Or, the neutral lipid is selected from phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, ceramide, a derivative thereof, and any combination thereof; Or, The neutral lipids include 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distaloyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-3-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether). PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero Lycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoylphosphatidylglycerol (DPPG), palmitoyloleoylphosphatidylethanolamine (POPE), distearoylphosphatidylethanolamine (DSPE), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), 1-stearoyl-2 - oleoyl-phosphatidylethanolamine (SOPE), 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine (LPE) and mixtures thereof; Or, 25. The composition of claim 24, wherein the neutral lipid is DOPE and / or DSPC.

26. The composition of claim 24, wherein the molar ratio of the cationic lipid, the neutral lipid, the structured lipid, and the polymer-conjugated lipid is (25-75):(5-25):(15-65):(0.5-10).

27. The composition of claim 26, wherein the molar ratio of the cationic lipid, the neutral lipid, the structured lipid, and the polymer-conjugated lipid is (35-49):(7.5-15):(35-55):(1-5).

28. 28. The composition of claim 27, wherein the molar ratio of the cationic lipid, the neutral lipid, the structured lipid, and the polymer-conjugated lipid is 49:10:43.5:1.

5.

29. 25. The composition of claim 24, wherein the structured lipid is selected from a sterol, cholesterol, a non-sterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid, α-tocopherol, a corticosteroid, and any combination thereof.

30. 30. The composition of claim 29, wherein the structured lipid comprises cholesterol.

31. 31. The composition of claim 30, wherein the structured lipid is cholesterol.

32. 25. The composition of claim 24, wherein the polymer-conjugated lipid is selected from PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and any combination thereof.

33. 33. The composition of claim 32, wherein the polymer-conjugated lipid is selected from distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), 1,2-dimyristoyl-rac-glycero-methoxypolyethylene glycol-2000 (DMG-PEG2000), methoxypoly(ethylene glycol) ditetradecylacetamide (ALC-0159), and combinations thereof.

34. The composition of claim 20, wherein the effective amount of VZV RNA is 25 μg to 200 μg.

35. The composition of claim 34, wherein the effective amount of VZV RNA is 50 μg to 100 μg.

36. 21. The composition of claim 20, wherein the vaccine is in an injectable form.

37. 37. The composition of claim 36, wherein the vaccine is a liquid formulation or a lyophilized formulation.

38. A method for preparing the composition of any one of claims 20 to 37, comprising the step of mixing the VZV RNA with the pharmaceutically acceptable carrier.

39. 39. A method for preparing the composition of claim 38, comprising encapsulating at least a portion of the RNA inside a lipid nanoparticle.

40. A medicament for inducing a protective immune response against VZV in a subject, comprising the composition of any one of claims 1 to 17 and 20 to 37.

41. 41. The composition of claim 40, wherein the protective immune response comprises the production of neutralizing antibodies.

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