PIV5-based antigen-modified vaccines: methods of making and using same

Abstract

The present invention relates to a recombinant vaccine comprising a PIV5-based viral expression vector with a genetic insert, and methods of producing and using the same. The recombinant vaccine comprises a modified bacterial or viral antigen to increase expression of the antigen on the surface of cells (AOS) to increase immunogenicity to the antigen.

Description

Technical Field

[0001] This invention generally relates to the field of vaccination, and more specifically, to compositions and methods for improving the immunogenicity of vaccine candidates by using a PIV5-based viral vector to express modified proteins to increase the expression of bacterial antigens on the cell surface (AOS). The antigen may be a bacterial antigen or a viral antigen. Background Technology

[0002] Parainfluenza virus type 5 (PIV5) belongs to the Paramyxoviridae family and the Mumpsvirus genus (mumps virus also belongs to this genus), and its genome is a single-stranded negative-sense RNA with a length of 15246 nt. The full-length genome structure of PIV5 is 3'-leader-NP-V / PMF-SH-HN-L-tail-5', that is, from the 3' end to the 5' end, excluding the leader and tail sequences, it encodes the nucleocapsid protein (NP), V protein / phosphorylated protein (P), matrix protein (M), fusion protein (F), small hydrophobic protein (SH), hemagglutinin-neuraminidase protein (HN), and polymerase protein (large protein, L) in sequence. Among them, V protein and SH protein are non-structural proteins.

[0003] PIV5 is an excellent viral vector for vaccine development, and research on recombinant PIV5 vaccines is currently underway. In recent years, researchers have continuously explored the feasibility of using PIV5 as a vaccine vector. A common approach is to insert protective antigen genes from viruses or bacteria into PIV5, and then express the inserted foreign gene through replication and translation of the PIV5 vector. Given that PIV5 can infect the respiratory tract without causing any disease, researchers often utilize this to focus on controlling certain respiratory viral infections. Therefore, in-depth research into the molecular biological characteristics and replication mechanisms of this virus is beneficial for a comprehensive and thorough understanding of PIV5, thus laying the foundation for research on PIV5 as a genetically engineered vaccine vector and for studying the gene function of this virus.

[0004] A notable characteristic of PIV5 is its ability to infect most mammalian cell types via sialic acid receptors without causing cytopathic effects. This allows PIV5 to replicate actively in the respiratory tract after intranasal immunization, inducing mucosal immunity by generating antigen-specific IgA antibodies and long-lived IgA plasma cells, as well as provoking humoral and cell-mediated immune responses throughout the body (Wang, D. et al., J Virol 91 (2017); Xiao, P. et al., Front Immunol 12, 623996 (2021)). Live PIV5 (non-replication-deficient type) has been administered intranasally to dogs as a component of kennel cough vaccines for over 50 years. Since the vaccine may shed within 5 days after administration (Chladek, DW, et al., American journal of veterinary research 42, 266-270 (1981)), humans have been exposed to PIV5 since the vaccine became available. However, no disease has been reported in exposed individuals, highlighting PIV5's excellent safety record. Previously, live PIV5 vector vaccines against bacterial pathogens such as Mycobacterium tuberculosis, Burkholderia melioides, and Burkholderia melioides have been tested in mouse models and demonstrated efficacy (Dyke, JS et al. Virulence 11, 1024-1040 (2020); Lafontaine, ER et al., Vaccine X 1, 100002 (2019); Chen, Z. et al. Vaccine 33, 7217-7224 (2015)). In addition, two PIV5 vector vaccines against COVID-19 and respiratory syncytial virus (RSV) (CVXGA1 and BLB201, respectively) have completed Phase I / II clinical trials.

[0005] This article provides a composition for a PIV5 vector vaccine that expresses modified bacterial or viral proteins to increase antigen expression on the cell surface (AOS). Methods for using this composition to enhance the immunogenicity of vaccine candidates are also provided. Summary of the Invention

[0006] According to the purposes of this invention, as practiced and extensively described herein, one aspect of the invention relates to a PIV5-based antigen expression vector, said vector comprising: a PIV5-based viral expression vector; and a gene for expressing an antigen in mammalian cells, wherein the gene expressing a bacterial antigen is modified by replacing the signal peptide of the gene with the N-terminal domain (NT) of a PIV5 HN protein or the N-terminal domain plus a transmembrane domain (NTTM) of a PIV5 HN protein, or by directly adding only the N-terminal domain (NT) of a PIV5 HN protein or adding the N-terminal domain plus a transmembrane domain (NTTM) sequence upstream of the antigen without removing the signal peptide; and wherein the gene expressing the modified bacterial or bacterial antigen is positioned between the SH and HN genes or between the HN and L genes in the PIV5 genome, or by replacing the SH gene. The antigen may be a viral antigen or a bacterial antigen.

[0007] In one embodiment, the PIV5-based viral expression vector comprises a PIV5-AVLP, PIV5-W3A, PIV5-CPI, or PIV5-CVB backbone. In another embodiment, the PIV5-based antigen expression vector has a heterologous nucleic acid sequence with at least 98% sequence identity to the sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.

[0008] In another embodiment, the modified antigen gene is expressed on the surface of mammalian cells. In another embodiment, the bacterial antigen is an antigen of Gram-negative bacteria selected from the group consisting of Burkholderia melioides, Borrelia burgdorferi, and Chlamydia trachomatis; and the viral antigen is an antigen of a virus selected from the group consisting of coronaviruses, respiratory syncytial virus (RSV), influenza A virus, influenza B virus, influenza C virus, orthomyxovirus, Sogoto virus, and Isa virus.

[0009] In one embodiment, the gene expressing the bacterial antigen is modified by replacing the N-terminal signal peptide of the *Burkholderia glandis* Pal protein with the N-terminal and transmembrane domain of the PIV5 HN glycoprotein. Another embodiment modifies the gene expressing the bacterial antigen by directly adding the N-terminal and transmembrane domain of the PIV5 HN glycoprotein upstream of the *Bretorius burgdorferi* OspA BPBPk protein. In yet another embodiment, the gene expressing the bacterial antigen is modified by replacing the signal peptide from the *Chlamydia trachomatis* major outer membrane protein (MOMP) with either the N-terminal domain of PIV5 HN alone or the N-terminal and transmembrane domain.

[0010] On the other hand, the present invention relates to a PIV5-based antigen vector composition, the composition comprising: a PIV5-based viral expression vector; and a gene for expressing an antigen in mammalian cells, wherein the PIV5-based viral expression vector comprises a PIV5-AVLP, PIV5-W3A, PIV5-CPI, or PIV5-CVB backbone; wherein the gene for expressing the bacterial antigen is modified by replacing the signal peptide of the gene with the N-terminal domain (NT) of the PIV5 HN protein or the N-terminal domain plus transmembrane domain (NTTM) of the PIV5 HN protein, or by directly adding only the N-terminal domain (NT) of the PIV5 HN protein or adding the N-terminal domain plus transmembrane domain (NTTM) sequence upstream of the antigen without removing the signal peptide; and wherein the gene expressing the modified antigen is positioned between the SH and HN genes or between the HN and L genes in the PIV5 genome, or by replacing the SH gene. The antigen may be a viral antigen or a bacterial antigen.

[0011] In some embodiments of the composition, the PIV5-based antigen vector has a nucleic acid sequence with at least 98% sequence identity to SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. In other embodiments, the composition is a vaccine against Gram-negative bacterial infection, wherein the Gram-negative bacteria are selected from the group consisting of Burkholderia melioides, Borrelia burgdorferi, and Chlamydia trachomatis; and the viral antigen is an antigen of a virus selected from the group consisting of coronaviruses, respiratory syncytial virus (RSV), influenza A virus, influenza B virus, influenza C virus, orthomyxovirus, Sogoto virus, and Isa virus.

[0012] In another aspect, the present invention relates to a method for inducing an immune response in a subject in need, the method comprising administering a PIV5-based antigen vaccine to the subject, wherein the immune response comprises a humoral immune response and / or a cellular immune response, and wherein the subject is susceptible to infection with Gram-negative bacteria selected from the group consisting of Burkholderia melioides, Borrelia burgdorferi, and Chlamydia trachomatis. In another embodiment, the subject is susceptible to viral infection, wherein the virus is selected from the group consisting of coronaviruses, respiratory syncytial virus (RSV), influenza A virus, influenza B virus, influenza C virus, orthomyxovirus, Sogoto virus, and Isa virus.

[0013] In some embodiments, the subject has previously been vaccinated against a bacterial or viral infection, and the method includes administering the composition to the subject. In other embodiments, the composition may be administered intranasally, intramuscularly, topically, or orally.

[0014] Other advantages of the invention will be set forth in part in the description which follows, and will be apparent in part from the description, or may be learned by practice of the invention. The advantages of the invention will be realized and achieved by means of the elements and combinations particularly pointed out in the appended claims. It should be understood that the foregoing general description and the following detailed description are exemplary and explanatory only, and do not limit the claimed invention. Attached Figure Description

[0015] The accompanying drawings, which are incorporated in and form part of this specification, illustrate several embodiments of the invention and, together with the description, serve to explain the principles of the invention.

[0016] Figure 1 A schematic diagram of PIV5-based constructs of Burkholderia melioides and Burkholderia pseudomelioides (Pal) cDNA is shown. PIV5-Pal contains the natural form of Pal, and PIV5-NTTM-Pal contains the N-terminus of PIV5 NH and a transmembrane domain (NTTM) added to the N-terminus of Pal.

[0017] Figure 2 Western blot analysis is shown to illustrate the expression of the Pal protein in cells infected with MHD11 (PIV5-Pal) and MHD39 (PIV5-NTTM-Pal) recombinant viruses and in the cell supernatant.

[0018] Figures 3A - 3B The PIV5 AVLP-based Burkholderia melioides and Burkholderia melioides (Pal) constructs and their immunogenicity are shown. Figure 3A This is a schematic diagram of the constructs of Burkholderia melioides and Burkholderia pseudomallei (Pal) cDNA based on PIV5 AVLP. PIV5 AVLP-Pal contains the natural form of Pal, and PIV5 AVLP-NTTM-Pal contains the N-terminus of PIV5 HN and a transmembrane domain (NTTM) added to the N-terminus of Pal. Figure 3B This is a graph showing the data on the anti-Pal serum IgG antibody response in AVLP-immunized mice. Five six- to eight-week-old female Balb / c mice were immunized with 3.8 x 10... 4Mice were immunized with either MHD30 or GOB10 AVLP or a PBS-only control. Animals were exsanguinated 14 days post-immunization, followed by a booster immunization with the same dose of each construct 21 days post-immunization. Mice were humanely euthanized 28 days post-immunization, and blood was collected. Serum samples were used for ELISA to quantify anti-Pal antibodies. GOD10 (AVLP-NTTM-Pal) elicited anti-Pal antibodies, while MHD30 (AVLP-Pal) did not.

[0019] Figures 4A - 4B A schematic diagram of pCAGGS and the cDNA of the PIV5-based Lyme disease (OspA / BPBPk) construct is shown. Figure 4A As shown, pMHD22 contains the human codon-optimized OspA BPBPk gene, and pMHD24 contains the human codon-optimized OspA BPBPk gene, with the addition of the PIV5 HN N-terminus and transmembrane domain (NTTM) at the N-terminus. Figure 4B The diagram shows that pMHD35 is PIV5 (W3A), containing a human codon-optimized OspA BPBPk gene, with a PIV5 HNN terminus and a transmembrane domain (NTTM) added to the N-terminus, and the chimeric gene inserted between the SH and HN genes; pMHD36 is PIV5 (W3A), containing a human codon-optimized OspA B31 gene, with a PIV5 HN N-terminus and a transmembrane domain (NTTM) added to the N-terminus, which is inserted between the SH and HN genes; pMHD106 is CPI, containing a human codon-optimized OspA BPBPk gene, with a PIV5 HN N-terminus and a transmembrane domain (NTTM) added to the N-terminus, which is inserted between the SH and HN genes; and pMHD116 is CVB, containing a human codon-optimized OspA BPBPk gene, with a PIV5 HN N-terminus and a transmembrane domain (NTTM) added to the N-terminus to replace the SH gene.

[0020] Figures 5A - 5B Western blot analysis of OspA expression in cell lysates of pMHD22 and pMHD24 transfected cells is shown. Figure 5A ) and bar chart ( Figure 5B ).

[0021] Figures 6A - 6C It was transfected with pMHD22 ( Figure 6B ) and pMHD24 ( Figure 6C Immunofluorescence analysis image of OspA expression on the surface of the construct cells. Images are shown at 20x magnification.

[0022] Figure 7The image shows the results of OspA expression detection using Western blot. Vero cells were subjected to simulated infection or infected with the PIV5 vector control at 1 MOI, and PIV5-A... B31 Or PIV5-A BPBPk Forty-eight hours post-infection, cells were lysed, and the lysate was separated on an SDS-PAGE gel and immunoblotted with anti-OspA and anti-PIV5 NP antibodies.

[0023] Figure 8 A graph showing serum IgG data for anti-OspA is presented. Mice were subcutaneously administered two doses of alum alone or alum + 20 μg rOspA. B31 Protein, or 10 nasal doses 6 PFU PIV5 vector control, PIV5-A BPBPk (MHD35) or PIV5-A B31 (MHD36), 21 days apart. Blood samples were collected on day 17 after primary immunization and at 3, 6, or 12 months post-immunization. Anti-OspA was quantified by ELISA. B31 IgG antibody titer. LOD is represented by a dashed line. Statistical significance of each vaccine group compared to pre-boost immunization levels was calculated using nonparametric tests and Dunn's multiple comparisons. (*p≤0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0024] Figures 9A - 9C This is a data graph showing the results of inoculation with PIV5-A. BPBPk Or PIV5-A B31 The neutralization of Borrelia burgdorferi by animal serum. On day 117, 4 months after the initial vaccination ( Figure 9A ), 270 days after the first vaccination (9 months) Figure 9B ) and 533 days after the first vaccination (18 months) Figure 9C Mice were challenged with ticks, and serum was collected from vaccinated mice afterward. Serum from the 4-month trial was pooled by group (n=5 mice / group) due to insufficient volume per individual. In subsequent trials (9 months and 18 months), serum from each group (n=3–6 mice) was tested individually. Statistical significance of each vaccine group compared to day 0 was calculated using nonparametric tests and Dunn's multiple comparisons. (*p≤0.05, **p<0.01, ****p<0.0001). *Bb*, *Borrelia burgdorferi*.

[0025] Figures 10A - 10F This is a data graph showing the results of PIV5-A vaccination. BPBPk Or PIV5-A B31The protective effect of the vaccine against multiple strains of Borrelia burgdorferi ticks in mice was investigated. All vaccinated mice were euthanized at 4, 9, and 15 months post-primary vaccination, and bladder, heart, and joint tissues were collected for Bb DNA purification and flaB qPCR. Bladder and heart tissues were also cultured in BSK-H medium to analyze Borrelia burgdorferi viability, and samples were collected for PCR confirmation. The Bb load in the tissues is shown. Figures 10A - 10C ) and the vitality of Bb in the organization ( Figures 10D - 10F A representative diagram of ). Bb, Borrelia burgdorferi.

[0026] Figure 11 This is a schematic diagram describing the cDNA of the pCAGGS Chlamydia (MOMP) construct. pCVL168 contains a codon-optimized MOMP gene, pCVL169 contains a human codon-optimized MOMP gene in which the signal peptide has been deleted and replaced with the CD5 signal peptide, pCVL170 contains a human codon-optimized MOMP gene in which a PIV5 HN N-terminal and transmembrane domain (NTTM) has been added to the N-terminus, pCVL171 contains a human codon-optimized MOMP gene in which the signal peptide has been deleted and replaced with a PIV5 HN N-terminal and transmembrane domain (NTTM), and pCVL172 contains a human codon-optimized MOMP gene in which the signal peptide has been deleted and replaced with a PIV5 HN N-terminal domain (NT).

[0027] Figure 12 This is a schematic diagram illustrating the cDNA of a PIV5-based Chlamydia trachomatis (MOMP) vaccine viral construct. pMHD175 is a CPI containing a human codon-optimized MOMP gene, where the signal peptide has been deleted and replaced with a CD5 signal peptide located between the SH and HN genes. pMHD177 is a CPI containing a human codon-optimized MOMP gene, where the signal peptide has been deleted and replaced with a PIV5 HN N-terminal domain (NT) located between the SH and HN genes. pMHD181 is a CVB containing a human codon-optimized MOMP gene, where the signal peptide has been deleted and replaced with a CD5 signal peptide instead of SH. pMHD182 is a CVB containing a human codon-optimized MOMP gene, where the signal peptide has been deleted and replaced with a PIV5 HN N-terminal domain (NT) instead of SH.

[0028] Figures 13A - 13F This is an immunofluorescence analysis image of total MOMP expression in cells transfected with the pCVL168-pCVL172 construct.

[0029] Figure 14This is a Western blot image of MOMP expression in cell lysates of cells transfected with the pCVL168-pCVL172 construct. Detailed Implementation

[0030] The invention can be more readily understood by referring to the following detailed description of preferred embodiments of the invention and the examples included therein, as well as by referring to the accompanying drawings and their preceding and following description.

[0031] I. Definition

[0032] To facilitate understanding of the principles and features of the various embodiments of this disclosure, various exemplary embodiments will be explained herein. While exemplary embodiments of this disclosure have been explained in detail, it should be understood that other embodiments are also covered. Therefore, the scope of this disclosure is not intended to be limited to the details of the construction and arrangement of the components set forth in the specification or embodiments. This disclosure can have other embodiments and can be practiced or implemented in various ways.

[0033] For clarity, specific terminology will be used in describing exemplary embodiments. Unless the context clearly requires otherwise, the singular forms “a,” “an,” and “the” as used in this specification and the appended claims include multiple indicators. For example, reference to a component is also intended to include a combination of multiple components. Reference to a composition containing “a” component is intended to include components other than those specified.

[0034] The range herein may be expressed as “about” or “approximately” or “generally” a particular value and / or to “about” or “approximately” or “generally” another particular value. When expressing such a range, other exemplary embodiments include from one particular value and / or to another particular value.

[0035] Similarly, as used in this article, expressions such as "substantially contains" or "substantially pure" can include "at least substantially contains" or "at least substantially pure" or "completely contains" or "completely pure".

[0036] In this document, the terms “patient,” “individual,” “subject,” and “animal” are used interchangeably and refer to mammals, including but not limited to humans and veterinary animals (e.g., cats, dogs, cows, horses, sheep, pigs, etc.) and laboratory animal models. In a preferred embodiment, the subject is a human.

[0037] As described herein, the term "vaccination" generally refers to the sequential administration of one or more antigens to a subject to produce and / or enhance an immune response against said antigens. Sequential administration includes primary immunization followed by one or more booster immunizations.

[0038] In the context of this invention, the term "pathogen" refers to any substance capable of causing a pathological condition. Examples of "pathogens" include, but are not limited to, cells (e.g., bacterial cells, diseased mammalian cells, cancerous mammalian cells), fungi, parasites, viruses, prions, or toxins. Preferred pathogens are infectious pathogens. In one specific embodiment, the infectious pathogen is a virus, such as coronaviruses, respiratory syncytial virus (RSV), human metapneumovirus (hMPV), orthomyxoviruses, influenza A viruses (including all strains with different HA and NA proteins, such as (non-limiting examples) H1N1, H1N2, H2N2, H3N2, H7N7, and H3N8); influenza B viruses, influenza C viruses, Sogoto viruses (including Dorivirus, Batken virus, SiAR 126 virus), chicken anemia virus, Isavirus (e.g., infectious salmon anemia virus), etc. In another embodiment, the infectious pathogen is a bacterial pathogen, such as Burkholderia melioides, Borrelia burgdorferi, Chlamydia trachomatis, Elizabethan meningitidis, etc.

[0039] As used herein, an antigen refers to any molecule capable of evoking an immune response from T cells or B cells in a subject. Pathogen-specific antigens are typically components obtained from or derived from said pathogen, containing epitopes and capable of evoking an immune response against said pathogen. Depending on the pathogenic material, antigens can be of various natures, such as (poly)peptides, proteins, nucleic acids, lipids, cells, etc. Attenuated live pathogens (e.g., bacteria, viruses) or their inactivated or inactivated forms, or substances purified therefrom, such as proteins, peptides, lipids, etc., may also be used. Antigens can be naturally occurring or artificially manufactured. They can be exogenous or endogenous in the treated mammal (e.g., tumor antigens). Antigens can be produced using techniques known in the art itself, such as synthetic or recombinant techniques or enzymatic methods.

[0040] In one specific embodiment, the antigen is a protein, polypeptide, and / or peptide. The terms “polypeptide,” “peptide,” and “protein” are used interchangeably herein to refer to a polymer of amino acid residues. These terms also apply to amino acid polymers, where one or more amino acid residues may be modified or non-naturally occurring residues, such as artificial chemical mimics of the corresponding naturally occurring amino acids. It should be understood that the term “protein” also includes fragments or variants of different antigens, such as fragments containing epitopes, or proteins obtained from pathogens that have subsequently been modified by enzymatic, chemical, mechanical, or thermal means.

[0041] Those skilled in the art will understand that any bacterial or viral antigen can be used in the system of the present invention to enhance the immune response against a desired antigen associated with that bacterial or viral antigen by expressing it as AOS. The scope of the invention also contemplates antigens that are not normally presented to the immune system, such as antigens in the cytosol, which, when used in conjunction with the system of the present invention, are also presented to the outer surface of viral particles, thereby enabling the immune system to generate an immunogenic response to these antigens that were previously “hidden” outside the immune system.

[0042] "Therapeutic effective amount" means the amount of a compound (e.g., the CVB-based composition described herein) that is sufficient to achieve therapeutic effect when administered to a subject in a therapeutic state, condition, or disease. "Therapeutic effective amount" can vary depending on the compound or bacteria administered, the disease and its severity, and the age, weight, physical condition, and responsiveness of the mammal being treated.

[0043] The phrase “pharmaceutical acceptable” as used with respect to compositions of this disclosure refers to physiologically tolerable molecular entities and other components of such compositions that generally do not produce adverse effects when administered to mammals (e.g., humans). Preferably, as used herein, the term “pharmaceutical acceptable” means approved by federal or state regulatory agencies or listed in the United States Pharmacopeia or other generally recognized pharmacopoeia for use in animals and more specifically in humans.

[0044] As used herein, the term "pharmaceuticalally acceptable composition" means a composition comprising at least one compound disclosed herein formulated together with one or more pharmaceutically acceptable carriers.

[0045] The term "administration" refers to the introduction of a predetermined amount of a substance into a patient's body by a suitable method. The compositions disclosed herein can be administered via any commonly used route, as long as it reaches the desired tissue, such as, but not limited to, inhalation, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, intrapulmonary, or rectal administration. However, because peptides are digested after oral administration, the active ingredient of compositions intended for oral administration should be coated or formulated to prevent degradation in the stomach.

[0046] The term "dosage" refers to a single amount of compound or agent administered to a patient; and / or "regimen" refers to multiple predetermined doses, which may differ or be similar in quantity, administered at different time intervals, which may differ or be similar in duration. In some embodiments, the regimen also includes the duration of a delivery period (e.g., administration period or treatment period). Alternatively, a regimen may be multiple predetermined evaporation doses administered at predetermined time intervals.

[0047] The term "carrier" refers to a diluent, adjuvant, excipient, or medium used to administer a compound. Such drug carriers can be sterile liquids such as water and oils, including those of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Water or aqueous saline solutions, as well as aqueous solutions of dextran and glycerol, are preferred as carriers, particularly for injectable solutions. Alternatively, carriers can be solid dosage form carriers, including, but not limited to, one or more binders (for pelleting), flow aids, encapsulating agents, flavoring agents, and coloring agents. Suitable drug carriers are described in EW Martin's "Remington's Pharmaceutical Sciences".

[0048] The term "treat" or "treatment" for a condition, symptom, or disease includes: (1) preventing or delaying the onset of at least one clinical or subclinical symptom of the condition, symptom, or disease in a subject who may have or is susceptible to the condition, symptom, or disease but has not yet developed or is exhibiting clinical or subclinical symptoms of the condition, symptom, or disease; or (2) suppressing the condition, symptom, or disease, i.e., preventing, reducing, or delaying the development of the disease or its recurrence (in the case of maintenance therapy) or at least one clinical or subclinical symptom of the disease; or (3) alleviating the disease, i.e. leading to the resolution of the condition, symptom, or disease or at least one clinical or subclinical symptom of the disease. The benefit is either statistically significant for the subject to be treated or at least perceptible to the patient or physician.

[0049] "Comprising," "containing," or "including" means that the composition, article, or method contains at least the said compound, ingredient, particle, or method step, but does not exclude the presence of other compounds, materials, particles, or method steps, even if such other compounds, materials, particles, or method steps have the same function as the said one.

[0050] As used herein, the term “parainfluenza virus 5 (PIV5)” includes, for example, but not limited to, strains KNU-11, CC-14, D277, 1168-1, and 08-1990. Non-restrictive examples of PIV5 genomes are listed in GenBank accessions NC_006430.1, AF052755.1, KC852177.1, KP893891.1, KC237065.1, KC237064.1, and KC237063.1, which are hereby incorporated by reference.

[0051] As used herein, the term "expression" refers to the process by which a polynucleotide is transcribed into mRNA and translated into a peptide, polypeptide, or protein. If the polynucleotide is derived from genomic DNA, then its expression may involve mRNA splicing, provided a suitable eukaryotic host cell or organism is selected. In the context of this invention, the term also includes the yield of the F gene mRNA and the F protein obtained after its expression.

[0052] As used herein, the terms “F protein” or “fusion protein” or “F protein polypeptide” or “fusion protein polypeptide” refer to a polypeptide or protein having all or part of the amino acid sequence of an RSV fusion protein polypeptide. Many RSV fusion proteins and attachment proteins have been described and are known to those skilled in the art. WO / 2008 / 114149 (incorporated herein by reference in its entirety) lists exemplary F protein and G protein variants (e.g., naturally occurring variants).

[0053] As used herein, the term "combination" in the context of a CVB-based composition with at least one second pharmaceutically active ingredient means at least two, but any desired combination of, compounds that can be delivered simultaneously or sequentially (e.g., within 24 hours). It is contemplated that the compositions and methods of this disclosure can be used in conjunction with other treatments / agents applicable to the same or similar diseases when used to treat various conditions. Such other treatments / agents can be co-administered (simultaneously or sequentially, in any order) to produce an additive or synergistic effect. Due to additive or synergistic effects, the appropriate therapeutically effective dose of each agent may be reduced. Furthermore, two or more embodiments of this disclosure can also be co-administered to produce an additive or synergistic effect.

[0054] As used herein, “Gram-negative bacteria” is a term used for the purposes of this specification and claims, and may, but is not limited to, describe one or more of the following bacterial species (i.e., combinations of one or more): Acinetobacter baumannii, Acinetobacter hemolyticus, Actinobacter actinomycetii, Aeromonas hydrophila, Bacteroides fragilis, Bacteroides polymorpha, Bacteroides ovalis, Bacteroides vulgaris, Bordetella pertussis, Borrelia burgdorferi, Brucella ovis, Burkholderia cepacia, Burkholderia cepacia, Burkholderia melioides, Burkholderia melioides, Prevotella humanis, Prevotella intermedius, Prevotella pulpis, Porphyromonas suisophilus, Campylobacter jejuni, Campylobacter coli, Campylobacter fetus, Citrobacter freundii, Citrobacter kurstii, Edwardsiella tarda, Ekenella erosus, Enterobacter cloacae, Enterobacter aerogenes, Enterobacter aggregates, Escherichia coli, Tula Francisella, Haemophilus influenzae, Haemophilus ducreyi, Helicobacter pylori, and Klebsiella kinesi. Bacteria, Klebsiella pneumoniae, Klebsiella oxytocinoides, Klebsiella rhinosclerotium, Klebsiella odorata, Legionella pneumophila, Moraxella catarrhalis, Morganella morganii, Neisseria gonorrhoeae, Neisseria meningitidis, Pasteurella multocida, Shigella-like bacteria, Proteus mirabilis, Proteus vulgaris, Proteus penebrion, Proteus myxobolus, Providencia squarrosa, Providencia retinae, Providencia alkaliformis, Pseudomonas aeruginosa, Pseudomonas fluorescens, Salmonella typhi. Salmonella paratyphi, Serratia marcescens, Shigella flexneri, Shigella boydii, Shigella sonnei, Shigella dysenteriae, Stenotrophomonas maltophilia, Streptococcus candida, Streptococcus pneumoniae, Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio alginolyticus, Yersinia enterocolitica, Yersinia plague, Yersinia pseudotuberculosis, Chlamydia pneumoniae, Chlamydia trachomatis, Rickettsia prowleri, Coxella burgdorferi, Ehrlichia chaffeensis, or Bartonella hensenae.

[0055] References to one or more method steps do not preclude the presence of additional or intermediate method steps between those explicitly identified steps. Similarly, it should be understood that references to one or more components in a composition do not preclude the presence of additional components besides those explicitly identified components.

[0056] The materials described to form the various elements of this disclosure are intended to be illustrative and not limiting. Many suitable materials that will perform the same or similar functions as the materials described herein are intended to be included within the scope of this disclosure. Such other materials not described herein may include, but are not limited to, materials developed, for example, after the time of development of this disclosure.

[0057] II. Composition

[0058] This article provides compositions using PIV5 (W3A, CPI, or CVB backbone) expression-modified bacterial proteins that increase antigen expression on the cell surface (AOS), thereby enhancing the immunogenicity of vaccine candidates.

[0059] A. Parainfluenza virus 5 (PIV5)

[0060] Parainfluenza virus 5 (PIV5) is a negative-sense RNA virus belonging to the genus Mumpsvirus in the family Paramyxoviridae. This genus includes many important human and animal pathogens, such as mumps virus, human parainfluenza virus types 2 and 4, Newcastle disease virus, Sendai virus, HPIV3, measles virus, canine distemper virus, rinderpest virus, and respiratory syncytial virus. PIV5 was formerly known as simian virus 5 (SV5). Although PIV5 is a virus that can infect many animals and humans, it has not yet been found to be associated with any known symptoms or diseases in humans. Unlike most paramyxoviruses, PIV5 exhibits little cytopathic effect when infecting normal cells. As a negative-sense RNA virus, the PIV5 genome is very stable. Because PIV5 lacks a DNA phase in its life cycle and replicates only in the cytoplasm, it cannot integrate into the host genome. Therefore, using PIV5 as a vector avoids potential unintended consequences due to genetic modification of the host cell's DNA. PIV5 can be grown at high titers in cells, including Vero cells, which have been approved by the WHO and FDA for vaccine production. Therefore, PIV5 has many advantages as a vaccine vector.

[0061] The PIV5 vaccine vector of the present invention can be constructed using any of a variety of methods, including but not limited to the reverse genetics system described in more detail in He et al. (Virology; 237(2):249-60, 1997). PIV5 encodes eight viral proteins. The nucleocapsid protein (NP), phosphoprotein (P), and large RNA polymerase (L) proteins are essential for the transcription and replication of the viral RNA genome. The V protein plays an important role in viral pathogenesis and viral RNA synthesis. The fusion (F) protein is a glycoprotein that mediates cell-cell and virus-cell fusion in a pH-independent manner, which is essential for viral entry into cells. The structure of the F protein has been determined, and key amino acid residues for efficient fusion have been identified. The hemagglutinin-neuraminidase (HN) glycoprotein is also involved in the process of viral entry into and release from host cells. The matrix (M) protein plays an important role in viral assembly and budding. The hydrophobic (SH) protein is a hydrophobic integrated membrane protein consisting of 44 residues, with its N-terminus located in the cytoplasm. For reviews on the molecular biology of paramyxoviruses, see, for example, Whelan et al., 2004, Curr Top Microbiol Immunol; 283:61-119; and Lamb & Parks, (2006). Paramyxoviridae: the viruses and their replication. In Fields Virology, 5th ed., pp. 1449–1496. Edited by DM Knipe & PM Howley. Philadelphia, PA: Lippincott Williams & Wilkins.

[0062] Previously, recombinant PIV5 viruses expressing foreign genes from a wide range of pathogens, including influenza virus, rabies virus, respiratory syncytial virus, tuberculosis virus, Burkholderia virus, and MERS-CoV virus, have been generated and tested as vaccine candidates (Li, Z., et al., J Virol, 87(1):354 (2013); Chen, Z., et al., J Virol, 87(6): 2986 (2013); Wang, D., et al., J Virol, 91(11) (2017); Chen, Z., et al., Vaccine, 33(51):7217 (2015); Lafontaine, ER, et al., Vaccine X., 1:100002 (2018); Li, K., et al., mBio, 11(2) (2020)). Because PIV5 vector vaccines actively replicate in the respiratory tract after intranasal immunization, they can generate mucosal immunity, including antigen-specific IgA antibodies and long-lived IgA plasma cells (Wang, D., et al., J Virol, 91(11) (2017). Xiao, P., et al., Front Immunol, 12:623996 (2021)). Recently, PIV5 vector vaccines expressing spike proteins from SARS-CoV-2 (WA1; CVXGA1) have been shown to be effective in mice and ferrets. A single intranasal dose of CVXGA1 induced WA1 neutralizing antibodies and protected K18-hACE2 mice from lethal infection with SARS-CoV-2 WA1. In addition, a single intranasal dose of CVXGA1 protected ferrets from infection with SARS-CoV-2 WA1 and blocked the transmission of the virus to uninfected ferrets in the same cage (An, D., et al., Sci Adv, 7(27) (2021)). While these studies have established its efficacy against SARS-CoV-2 WA1, further research is needed to determine its efficacy against SARS-CoV-2 variants.

[0063] 1. PIV5 - W3A

[0064] In one embodiment, the PIV5-based vaccine vector of the present invention is based on any of a variety of wild-type, mutant, or recombinant (rPIV5) strains. Wild-type strains include, but are not limited to, PIV5 strains W3A, WR (ATCC® number VR-288TM), canine parainfluenza virus strain 78-238 (ATCC number VR-1573) (Evermann et al., 1980, J Am VetMed Assoc; 177:1132-1134; and Evermann et al., 1981, Arch Virol; 68:165-172), canine parainfluenza virus strain D008 (ATCC number VR-399) (Binn ​​et al., 1967, Proc Soc Exp Biol Med; 126:140-145), MIL, DEN, LN, MEL, cryptovirus, CPI+, CPI-, H221, 78524, T1, and SER. See, for example, Chatziandreou et al., 2004, J Gen Virol; 85(Pt 10):3007-16; Choppin, 1964, Virology: 23:224-233; and Baumgartner et al., 1987, Intervirology; 27:218-223. Alternatively, the PIV5 strain used in commercial kennel cough vaccines, such as BI, FD, Merck, and Merial vaccines, can be used.

[0065] 2. PIV5 – CP1

[0066] In another embodiment, the PIV5-based vaccine vector of the present invention is based on the PIV5 CPI strain vector backbone. The most significant difference between the PIV5 CPI and PIV5 W3A strain backbones is that the CPI strain has an additional 22 amino acid extension in the cytoplasmic tail region of the PIV5 F protein. This F protein extension is believed to inhibit viral fusion properties. Compared to W3A-based viruses, CPI-based viruses exhibit stronger lytic activity and produce more progeny viruses in infected cells, whereas W3A-based viruses lack the extended PIV5 F protein tail and exhibit additional amino acid differences compared to CPI.

[0067] 3. PIV5-CVB

[0068] In yet another embodiment, the PIV5 virus vaccine of the present invention will have mutations, alterations, or deletions in one or more of the eight proteins of the PIV5 genome. For example, the PIV5 virus expression vector may contain one or more mutations, including but not limited to any of those mutations described herein. In some aspects, a combination of two or more (two, three, four, five, six, seven or more) mutations may be advantageous and may exhibit enhanced activity.

[0069] In one implementation, the PIV5 vector is further modified by introducing a mutation into the PIV5 V / P gene and deleting the PIV5 SH gene, thereby further improving vaccine efficacy. Previous studies have shown that S157F and S308A in the PIV5 V / P gene can enhance viral polymerase activity and increase viral titer or yield (Timani KA, Sun D, ​​Sun M, et al. JVirol., 82(18):9123-9133 (2008); Sun D, ​​Luthra P, Li Z, He B., PLoS Pathog., 5(7):e1000525 (2009)). In rats, RSV vaccines based on the PIV5 W3A strain containing a single S157F mutation induced higher levels of neutralizing antibodies than PIV5 CPI vector RSV vaccines (see Table 19). The PIV5 W3A strain, lacking the SH gene and expressing the influenza virus H5 HA protein, induces higher levels of antibodies and provides better protection against influenza virus attack (Li Z, Gabbard JD, Mooney A, et al., J Virol., 87(17): 9604–9609 (2013)). In one embodiment, this paper provides a newly generated modified PIV5 viral vector backbone, named CVB, by introducing S157F into the V / P gene and deleting the SH gene from the PIV5 W3A viral genome. In another embodiment, a CVB-vectored SARS-CoV-2 vaccine for intranasal immunization was generated.

[0070] Mutations include, but are not limited to, V / P gene mutations, mutations at the N-terminus shared by V and P proteins, mutations at residues 26, 32, 33, 50, 102, 156 and / or 157 at the N-terminus shared by V and P proteins, mutations lacking the C-terminus of V protein, mutations lacking small hydrophobic (SH) protein, fusion (F) protein mutations, phosphoprotein (P) mutations, large RNA polymerase (L) protein mutations, mutations incorporating canine parainfluenza virus residues, and / or mutations enhancing syncytial formation.

[0071] Mutations may include, but are not limited to, rPIV5-V / P-CPI-, rPIV5-CPI-, rPIV5-CPI+, rPIV5V ΔC, rPIV-Rev, rPIV5-RL, rPIV5-P-S156N, rPIV5-P-S157A, rPIV5-P-S308A, rPIV5-L-A1981D, rPIV5-F-S443P, rPIV5-MDA7, rPIV5 ΔSH-CPI-, rPIV5 ΔSH-Rev, and combinations thereof.

[0072] PIV5 can efficiently infect cells in many cell types with minimal cytopathic effect (CPE). In some cell types, PIV5 infection leads to syncytial formation, where many cells fuse together, resulting in cell death. Mutations can include one or more mutations that promote syncytial formation (see, for example, Paterson et al., 2000, Virology; 270:17-30).

[0073] The V protein of PIV5 plays a crucial role in blocking virus-induced apoptosis. Recombinant PIV5 lacking the conserved cysteine-rich C-terminus (rPIV5V ΔC) of the V protein induces apoptosis in various cell types via an intrinsic apoptotic pathway, possibly triggered by endoplasmic reticulum (ER) stress (Sun et al., 2004, J Virol; 78:5068-5078). Mutant recombinant PIV5 with mutations at the N-terminus of the V / P gene product, such as rPIV5-CPI-, also induces apoptosis (Wansley and Parks, 2002, J Virol; 76:10109-10121). Mutations include, but are not limited to, rPIV5 ΔSH, rPIV5-CPI-, rPIV5VΔC, and combinations thereof.

[0074] 4. PIV5-AVLP

[0075] This disclosure provides PIV5-based AVLP compositions and methods for use as bacterial vaccines. This disclosure relates to PIV5-based AVLP compositions and constructs for their manufacture, which can be used to introduce an expressible target polynucleotide sequence into host cells. In some embodiments, the PIV5-based AVLP composition is an isolated polynucleotide sequence that transcribes a single-stranded RNA encoding a portion of the negative-strand PIV5 genome, wherein the polynucleotide sequence transcribes at least a portion of the negative-strand NP, V / P, and L genes, and wherein the polynucleotide sequence lacks or is unable to transcribe one or more of the M, F, SH, and HN genes, and wherein the polynucleotide sequence contains a heterologous non-PIV5 nucleotide sequence inserted between the V / P and L genes (e.g.,Figure 1 A). In some embodiments, one or all of the M, F, SH, and HN genes are completely removed and replaced with an expressible target heteronucleotide sequence. In some embodiments, the isolated polynucleotide contains all of the NP, V / P, and L genes. In some embodiments, the isolated polynucleotide contains all of at least one of the NP, V / P, and L genes.

[0076] In some embodiments, the PIV5-based AVLP composition comprises at least one AVLP particle containing isolated polynucleotides as described herein. In other embodiments, the PIV5-based AVLP composition comprises a plurality of AVLP particles containing isolated polynucleotides as described herein.

[0077] The compositions disclosed herein can be formulated into various forms of pharmaceutical preparations to suit a chosen route of administration. Those skilled in the art will understand that the compositions will vary depending on the method of administration and dosage unit.

[0078] The agents of the present invention can be administered in a variety of ways, including but not limited to intravenous, local, oral, intranasal, subcutaneous, intraperitoneal, intramuscular, and intratumoral delivery. In some aspects, the agents of the present invention can be formulated for controlled or sustained release. One advantage of intranasal immunization is the potential to induce a mucosal immune response.

[0079] B. Bacterial antigens expressed by the PIV5 vector

[0080] This document provides compositions in which a PIV5, AVLP, CPI, or CVB backbone is used as a viral vector to express bacterial antigens on the surface of virus-infected cells by introducing a PIV5HN N-terminal cytoplasmic tail region or a cytoplasmic tail region with a transmembrane domain. This invention overcomes the difficulties in displaying bacterial antigens on cell surfaces and improves antigen-specific immune responses. Three exemplary bacterial antigens successfully expressed by the PIV5 vector are listed here: Burkholderia melioides Pal protein, Lyme disease (Breorhizium burgdorferi) OspA protein, and Chlamydia trachomatis MOMP protein.

[0081] 1. Burkholderia melioides Pal protein

[0082] Lipopolysaccharide (LPS, an endotoxin) is a bacterial component released by Gram-negative bacteria and has been shown to play an important role in inducing sepsis. An early, pioneering study showed that polyclonal antiserum against heat-inactivated *E. coli* J5 (with an exposed LPS core) in humans reduced GNS-related mortality by half. Subsequent studies showed that antibodies targeting only the LPS core were not protective. It was later discovered that IgG in the J5 antiserum could bind to three *E. coli* outer membrane proteins: Lpp, OmpA, and peptidoglycan-associated lipoprotein (Pal). Since these studies, in vitro and in vivo results have further demonstrated the association of Pal with the pathology of GNS.

[0083] Burkholderia melioides is a host-adapted Gram-negative bacterium that causes melioidosis, a highly fatal zoonotic disease that primarily affects horses, donkeys, and mules. The organism is highly contagious through contact with infected equines and is also susceptible to infection in humans (Khan I, et al., Transbound Emerg Dis. 60(3):204–221 (2013)).

[0084] Comparative analysis indicates that *Burkholderia melioides* evolved from *Burkholderia melioides* through genome reduction. The latter is common in water and moist soil in equatorial countries and causes the emerging tropical disease melioidosis globally (Wiersinga WJ, et al., *Nat Rev Dis Primers* 4:17107 (2018); Limmathurotsakul D, G, et al., *Nat Microbiol.* 1:1 (2016)). The genes retained by *Burkholderia melioides* show high sequence identity with their orthologs, and the two species share many virulence determinants. The clinical and pathological manifestations of the diseases caused by these organisms are also very similar. In humans, infection typically occurs through broken skin and the respiratory tract, most commonly manifesting as life-threatening pneumonia and bacteremia (Carr-Gregory B, Waag DM. Glanders. In: Dembek ZF, ed. Medical aspects of biological warfare. Borden Institute, Office of the Surgeon General, AMEDD Center and School, Texas, US; 2007. pp. 121–146.; Wiersinga WJ, et al., Nat Rev Dis Primers 4:17107 (2018)). Pathogenesis is complex, involving the simultaneous expression of genes supporting intracellular and extracellular bacterial replication, colonization of deep tissues and organ systems, and the development of a hallmark chronic condition that is difficult to treat and eliminate.

[0085] Peptidoglycan-associated lipoproteins (Pal) are highly conserved in Enterobacteriaceae but are also found in most Gram-negative bacteria. Pal is a component of the Tol-Pal system. These proteins form a complex that stretches the inner and outer membranes of Gram-negative bacteria and plays a role in maintaining cell wall integrity (Dyke, JS et al., Virulence 11, 1024–1040 (2020)). Pal can be used to access macromolecules such as bacteriocins and bacteriophages (Godlewska R et al., FEMS Microbiol Lett. 298(1):1–11 (2009); Duche D, EcoSal Plus. 8:2 (2019)).

[0086] 2. Lyme disease (Borrelia burgdorferi) OspA protein

[0087] Lyme disease is a tick-borne disease caused by *Borrelia burgdorferi* (SL). The disease is characterized by an enlarging red rash at the tick bite site, which may be followed by systemic complications including meningitis, myocarditis, or arthritis. Almost all cases of Lyme disease are caused by one of three genotypes: *Borrelia auriculata*, *Borrelia galbana*, and *Borrelia burgdorferi* (SS). In Europe, all three species have been found to infect humans. However, in North America, only one species, *Borrelia burgdorferi*, has been found. *Borrelia burgdorferi* is a Gram-negative bacterium belonging to the genus *Borrelia* in the class Spirochetes. Antibiotic treatment for Lyme disease is usually effective, but some patients develop a chronic, disabling disease affecting the joints or nervous system that does not significantly improve even with parenteral antibiotic therapy, highlighting the need for vaccination for high-risk populations.

[0088] Outer surface protein A (OspA) is a 31 kDa antigen expressed by the species *Borrelia burgdorferi* s. / ., which is present in the midgut of hard ticks. OspA has been shown to be effective in preventing Lyme disease in North America (Steere et al., N. Engl. J. Med. 339: 209-15, 1998; Sigal et al., N. Engl. J. Med. 339: 216-22, 1998; erratum in: N. Engl. J. Med. 339: 571, 1998). The fully processed OspA has a cysteine ​​residue at its amino terminus, which is modified post-translation by three fatty acyl chains that anchor the protein to the outer surface of the bacterial membrane (Bouchon et al., Anal. Biochem. 246: 52-61, 1997). It has been reported that lipidation of OspA stabilizes the molecule (US8623375B2) and is crucial for protection in the absence of strong adjuvants (Erdile et al., Infect. Immun. 61: 81-90, 1993). The structure of OspA was determined by co-crystallizing a soluble recombinant form lacking an amino-terminal lipid membrane anchor with a Fab fragment of an agglutinating mouse monoclonal antibody. The results showed that OspA contains 21 antiparallel β-sheets and a single α-helix (Li et al., Proc. Natl. Acad. Sci. USA 94:3584-9, 1997).

[0089] A monovalent OspA-based vaccine (LYMErix®) is available in the United States for the prevention of Lyme disease. However, in Europe, OspA sequences are heterogeneous across the three gene strains, thus single-strain OspA-based vaccines do not provide broad protection (Gern et al., Vaccine 15:1551-7, 1997). Seven major OspA serotypes have been identified in European isolates (designated serotypes 1 to 7, Wilske et al., J. Clin. Microbiol. 31:340-50, 1993). OspA serotypes are species-related; serotype 1 corresponds to *Borrelia burgdorferi* ss, serotype 2 to *Borrelia auriculata*, and serotypes 3 to 7 to *Borrelia gargiensis*.

[0090] Protective immunity acquired through OspA immunization is unusual because the interaction between the host's immune response and the pathogen occurs not within the host, but rather within the midgut of the tick vector. In the case of Lyme disease, ticks are the vectors or carriers that transmit Lyme disease from animals to humans. OspA-specific antibodies acquired by infected ticks during blood-feeding can prevent the transmission of Borrelia burgdorferi sl to immune mammalian hosts (de Silva et al., J. Exp. Med. 183:271-5, 1996). The protective effect is antibody-mediated, primarily acting through bactericidal antibodies, although antibodies that block spirochetal attachment to the tick's intestinal epithelial lining receptors may also be effective (Pal et al., J. Immunol. 166: 7398-403, 2001).

[0091] The rational development of effective OspA vaccines requires the identification of protective epitopes, such as those defined by the protective monoclonal antibody LA-2 (Golde et al., Infect. Immun. 65: 882-9, 1997). X-ray crystallography and NMR analysis have been used to identify immunologically important hypervariable domains in OspA and to localize the LA-2 epitope to amino acids 203-257 (Ding et al., J. Mol. Biol. 302: 1153-64, 2000; Luft et al. J Infect Dis. 185(Suppl. 1): S46-51, 2002).

[0092] 3. Chlamydia trachomatis MOMP protein

[0093] Chlamydia is a prevalent sexually transmitted infection affecting more than 100 million people worldwide. While most individuals infected with Chlamydia trachomatis are initially asymptomatic, symptoms may develop if not diagnosed promptly. Long-term infection can lead to debilitating side effects such as pelvic inflammatory disease, infertility, and blindness. Therefore, Chlamydia infection poses a significant public health threat and underscores the need for a vaccine.

[0094] Chlamydia strains express a major outer membrane protein (MOMP), which has proven to be an effective vaccine antigen. However, the production of functional recombinant MOMP proteins for vaccine development has been challenging due to their poor solubility, low yield, and protein misfolding.

[0095] Chlamydia are obligate intracellular pathogens of eukaryotic cells. Currently, four species of Chlamydia are known—Chlamydia trachomatis, Chlamydia pneumoniae, Chlamydia sheep, and Chlamydia psittaci—and the genome sequences of each species are publicly available ((1999) Nature Genetics 21:385-389; (2000) Nucleic Acids Res 28:1397-1406; (2000) Nucleic Acids Res 28:2311-2314; (1998) Science 282:754-759).

[0096] Chlamydia trachomatis organisms exist in two states, alternating between two forms: 1) the infectious "epithecus" (EB), which is endocytosed by mucosal cells to form vesicle inclusion bodies; and 2) the metabolically active intracellular "reticulum" (RB). RB replicates and redifferentiates into EB, which is then released to infect neighboring cells.

[0097] Treatment of EB with n-lauroyl sarcosine produces the "Chlamydia OM complex" (COMC), which contains three relatively detergent-resistant, cysteine-rich proteins: the major outer membrane protein (MOMP) encoded by ompA, and OmcB and OmcA encoded by omp2 and omp3, respectively (BMC Microbiology 2005, 5:5, and references therein). There are also many other Chlamydia outer membrane proteins, such as PorB, PmpB, PmpC, PmpD, PmpG, and PmpH.

[0098] MOMP is expressed in both EB and RB and is located on the outer membrane, functioning as a porin. It accounts for approximately 60% of the membrane proteins in infectious EB. Structural and functional analyses show that native MOMP exists in oligomeric form; the native conformation of Chlamydia trachomatis MOMP is trimer, with its monomers possessing a β-barrel, β-sheet secondary structure. In EB, MOMP forms trimers, with disulfide bridges existing within and between its monomers (~40 kDa) and between trimers (J. Bacterid. 189:6222-6235, 2007).

[0099] At least 19 different Chlamydia trachomatis serotypes can infect humans (i.e., A to K, Ba, Da, La, Ja, L1 to L3 and L2a), and these serotypes are classified based on serological differences in antigenic epitopes on the MOMP. The MOMP encoded by each of these 19 different serotypes contains five highly conserved regions and four variable sequence segments or domains (referred to as VS or VD1 to VD4). Subspecies- and serotype-specific antigenic epitopes or determinants are located on the variable domains. Based on amino acid homology, serotypes are further subdivided into the following serogroups or categories: class B (B, Ba, D, Da, E, L1, L2, and L2a), class C (A, C, H, I, La, J, K, and L3), and intermediate classes (F and G). Infection with any Chlamydia trachomatis serotype can lead to disease: serotypes A, B, Ba, and C cause trachoma; serotypes L1-L3 are the pathogens of lymphogranuloma venereum; serotype DK causes sexually transmitted infections; and serotypes G, I, and D are associated with cervical cancer.

[0100] C. Viral antigens expressed by the PIV5 vector

[0101] Vaccines typically contain multiple antigenic components, such as those derived from different proteins and / or different epitope regions of the same protein. For example, a vaccine against a viral disease may contain one or more polypeptide sequences derived from a virus, which, when administered to a host, can elicit an immunogenic or protective response against viral attack.

[0102] As previously described, this disclosure can also be used to prepare polypeptide multimers, for example, wherein antigenic preparations composed of more than one polypeptide are produced. For example, a viral capsid can be composed of more than one polypeptide subunit. By transducing host cells with vectors carrying different viral envelope sequences, when proteins are expressed in the cells, these proteins can self-assemble into three-dimensional structures containing more than one protein subunit (e.g., in their native conformation).

[0103] In other embodiments, the expressible heterologous nucleotide sequence is derived from another virus besides PIV5. For example, the heterologous nucleotide sequence may encode influenza HA, RSV F, HIV Gag, and / or Env (from any strain). Such embodiments can be used to develop vaccines and / or vaccination methods. The examples given herein are non-limiting, as those skilled in the art will understand that nucleotide sequences from a variety of pathogens (including bacteria, parasites, etc.) may be suitable for AVLP vaccine compositions and / or vaccination methods.

[0104] Examples of viruses from which vaccines can be generated according to this disclosure include, for example, orthomyxoviruses, influenza A viruses (including all strains with different HA and NA proteins, such as (non-limiting examples) H1N1, H1N2, H2N2, H3N2, H7N7, and H3N8); influenza B viruses, influenza C viruses, sogoto viruses (including dorivirus, batken virus, and SiAR 126 virus), respiratory syncytial virus (RSV), and issaviruses (e.g., infectious salmon anemia virus), coronaviruses, etc. These include influenza viruses isolated or transmitted from all species types, including viruses isolated from invertebrates, vertebrates, mammals, humans, non-human primates, monkeys, pigs, dairy cows and other livestock, birds, poultry (such as turkeys, chickens, quails, and ducks), wild birds (including waterfowl and landfowl), reptiles, etc. These also include existing strains that have been altered, for example through mutation, antigenic drift, antigenic conversion, recombination, etc., especially strains that have been enhanced in virulence and / or spread across species (e.g., human-to-human transmission).

[0105] D. Composition sequence

[0106] 1. PIV5 vaccine sequence

[0107] a. pMHD11- PIV5-Pal

[0108] The pMHD11-PIV5-Pal nucleic acid sequence provided in this article, from 5' to 3', is as follows:

[0109] ACCAAGGGGAAAATGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCT ATGGCCTTCGTGACCGACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCGA CACAAAAATGTCATCCGTGCTTAAAGCATATGAGCGATTCACGCTCACTCAAGAACTGCAAGATCAGAGTGAGGAA GGTACAATCCCACCTACAACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGAT CCCGGCTTCTTCTATTCTGCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACT CACAATGTTTTCGCTACCATCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATC GAAAGGGTAGAGATCGATGGCTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCTCGTTCAGGTATGAGCCGTG GAGAGATCAATGCCTATGCTGCACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTCGTTGATTC CGAAGTCGAGGGAACTGCATGGGATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGG ATAGTGACTTGCAAGTGCATGACTGCGCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAATATCGTC AGCAAGGCAGGATCAACCCGAGATATCTCCTGCAACCGGAGGCTCGACGAATAATCCAGAATGTAATCCGGAAGGG AATGGTGGTCAGACATTTCCTCACCTTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTAT GCTATGGTAGGGGATGTTGGAAAGTATATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAAATATGCAT TAGGAACTAGATGGCCCACACTTGCTTTAGCTGCATTTTCAGGAGAGCTAACAAAGCTAAAGTCCTCATGGCATT ATACCAGACCCTTGGTGAGCAGGCCCGATATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTGCTGCAGCA AACTACCCACTGCTATATAGCTATGCTATGGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTTCT CCAGATCATACATGAACAAGACATATTTCCAATTGGGAATGGAACTGCAAGAAAACAACAGGGTGCAGTTGACAT GAGGATGGCAGAAGATCTCGGTCTAACTCAAGCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACA GCAAATCGAGGGGCAGACACCAGGGGAGGAGTCAACCCGTTCTCATCTGTCACTGGGACAACTCAGGTGCCCGCTG CAGCAACAGGTGACACACTCGAGAGTTACATGGCAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCA TGATGATGAGATGCCACCATTGGAAGAGGAAGAGGACGACACATCTGCAGGTCCACGCACTGGACCAACTCTT GAACAAGTGGCCTTGGACATCCAGAACGCAGCAGTTGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGG GTGATCTTGACATCTAGACAAATTCAGATCCCAATCTAAAATTGACATACCTAATTGATTAGTTAGATGGAACTACA GTGGATTCCATAAGGTTCCTGCCTACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGAC TGCCGGTGCCAACAGCGCAATCCACAATCTACAATGGATCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATA AGCTCATAGAGACAGGCCTGAATACTGTAGGTATTTTACTTCCCAACAAGTCACAGGAACATCCCTCTTTGGAAA GAATACAATACCACCAGGGGTCACAGGACTACTAACCAATGCTGCAGAGGCAAAGATCCAAGAGTCAACTAACCAT CAGAAGGGCTCAGTTGGTGGGGGGTGCAAACCAAAGAAACCGCGACCAAAAATTGCCATTGTGCCAGCAGATGACA AAACAGTGCCCGGAAAGCCGATCCCAAACCCTCTTATTAGGTCTGGACTCCACCCCGAGCACCCAAACTGTGCTTGA TCTAAGTGGGAAACATTACCATCAGGATCCTATAAGGGGGTTAAGCTTGCGAAATTTGGAAAGAAAATCTGATG ACACGGTTCATCGAGGAACCCAGAGAGAATCCTATCGCAACCAGTTCCCCCATCGATTTTAAGAGGGGCAGGGATA CCGGCGGGTTCCATAGAAGGGAGTACTCAATCGGATGGGTGGAGATGAAGTCAAGGTCACTGAGTGGTGCAATCC ATCCTGTTTCTCCAATCACCGCTGCAGCAAGGCGATTTGAATGCACTTGTCACCAGTGTCCAGTCACTTGCTCTGAA TGTGAACGAGATACTTAATACAGTGAGAAATTTGGACTTCCGGATGAATCAACTGGAGACAAAAGTAGATCGCATT CTTCCATCTCAGTCTCTAATCCAGACCATCAAGAATGACATAGTTGGACTTAAAGCAGGGATGGCTACTTTAGAAG GAATGATTACAACTGTGAAAATCATGGACCCGGGAGTTCCCAGTAATGTTACTGTGGAAGATGTACGCAAGACACT AAGTAACCATGCTGTTGTTGTCCAGAATCATTCCAATGATAGTTTCTTGACTCAATCTGAAGATGTAATTTCACTT GATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAGAAGATTGTCAGGAAGGTTCCTCCTCAGAAGGATCTGACTG GATTGAAGATTACACTAGAGCAATTGGCAAAGGATTGCATCAGCAAACCGAAGATGAGGGAAGAGTATCTCCTCAA AATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCTCAAGAAAGCAATCATCCGCAGTGCAATTTGATCAAGA AACACCCAATTACACTACACTGGTATGACACTGTACTAACCCTGAGGGTTTTAGAAAAAACGATTAACGATAAATA AGCCCGAACACTACACACTACCTGAGGCAGCCATGCCATCCATCAGCATTCCCGCAGACCCCACCAATCCACGTCA ATCAATAAAAGCGTTCCCAATTGTGATCAACAGTGATGGGGGTGAGAAAGGCCGCTTGGTTAAACAACTACGCACA ACCTACTTGAATGACCTAGATACTCATGAGCCACTGGTGACATTCATAAATACCTATGGATTCATCTACGAACAGG ATCGGGGGAATACCATTGTCGGAGAGGATCAACTTGGGAAGAAAAGAGAGGCTGTGACCGCTGCAATGGTTACCCT TGGATGTGGGCCTAATCTACCATCATTAGGGAATGTCCTGGGACAACTGAGGGAATTCCAGGTCACTGTTAGGAAG ACATCCAGCAAAGCGGAAGAGATGGTCTTTGAAATTGTTAAGTATCCGAGAATATTTCGGGGTCATACATTAATCC AGAAAGGACTAGTCTGTGTCTCCGCAGAAAAATTTGTTAAGTCACCAGGGAAAATACAATCTGGAATGGACTATCT CTTCATTCCGACATTTCTGTCAGTGACTTACTGTCCAGCTGCAATCAAATTTCAGGTACCTGGCCCCATGTTGAAA ATGAGATCAAGATACACTCAGAGCTTACAACTTGAACTAATGATAAGAATCCTGTGTAAGCCCGATTCGCCACTTA TGAAGGTCCATACCCCTGACAAGGAGGGAAGAGGATGTCTTGTATCAGTATGGCTGCATGTATGCAACATCTTCAA ATCAGGAAACAAGAATGGCAGTGAGTGGCAGGAATACTGGATGAGAAAGTGTGCTAACATGCAACTTGAAGTGTCG ATTGCAGATATGTGGGGACCAACTATCATAATTCATGCCAGAGGTCACATTCCCAAAAGTGCTAAGTTGTTTTTTG GAAAGGGTGGATGGAGCTGCCATCCACTTCACGAAGTTGTTCCAAGTGTCACTAAAACACTATGGTCCGTGGGCTG TGAGATTACAAAGGCGAAGGCAATAATACAAGAGAGTAGCATCTCTCTTCTCGTGGAGACTACTGACATCATAAGT CCAAAAGTCAAAATTTCATCTAAGCATCGCCGCTTTGGGAAATCAAATTGGGGTCTGTTCAAGAAAACTAAATCAC TGCCTAACCTGACGGAGCTGGAATGACTGACCTCTAATCGAGACTACACCGCCGCAAACTATAGGTGGGTGGTACC TCAGTGATTAATCTTGTAAGCACTGATCGTAGGCTACAACACACTAATATTATCCAGATTAGAGAGCTTAATTAGC TCTGTATTAATAATAACACTACTATTCCAATAACTGGAATCACCAGCTTGATTTATCTCCAAAATGATTCAAAGAA AACAAATCATATTAAGACTATCCTAAGCACGAACCCATATCGTCCTTCAAATCATGGGTACTATAATTCAATTTCT GGTGGTCTCCTGTCTATTGGCAGGAGCAGGCAGCCTTGATCCAGCAGCCCTCATGCAAATCGGTGTCATTCCAACA AATGTCCGGCAACTTATGTATTATACTGAAGCTTCATCAGCATTCATTGTTGTGAAGTTAATGCCTACAATTGACT CGCCGATTAGTGGATGTAATATAACATCAATTTCAAGCTATAATGCAACAGTGACAAAACTCCTACAGCCGATCGG TGAGAATTTGGAGACGATTAGGAACCAGTTGATTCCAACTCGGAGGAGACGCCGGTTTGCAGGGGTGGTGATTGGA TTAGCTGCATTAGGAGTAGCTACTGCCGCACAGGTCACTGCCGCAGTGGCACTAGTAAAGGCAAATGAAAATGCTG CGGCTATACTCAATCTCAAAAATGCAATCCAAAAAACAAATGCAGCAGTTGCAGATGTGGTCCAGGCCACACAATC ACTAGGAACGGCAGTTCAAGCAGTTCAAGATCACATAAACAGTGTGGTAAGTCCAGCAATTACAGCAGCCAATTGT AAGGCCCAAGATGCTATCATTGGCTCAATCCTCAATCTCTATTTGACCGAGTTGACAACCATCTTCCACAATCAAA TTACAAACCCTGCATTGAGTCCCATTACAATTCAAGCTTTAAGGATCCTACTGGGGAGTACCTTGCCGACTGTGGT CGAAAAATCTTTCAATACCCAGATAAGTGCAGCTGAGCTTCTCTCATCAGGGTTATTGACAGGCCAGATTGTGGGA TTAGATTTGACCTATATGCAGATGGTCATAAAAATTGAGCTGCCAACTTTAACTGTACAACCTGCAACCCAGATCA TAGATCTGGCCACCATTTCTGCATTCATTAACAATCAAGAAGTCATGGCCCAATTACCAACACGTGTTATGGTGAC TGGCAGCTTGATCCAAGCCTATCCCGCATCGCAATGCACCATTACACCCAACACTGTGTACTGTAGGTATAATGAT GCCCAAGTACTCTCAGATGATACTATGGCTTGCCTCCAAGGTAACTTGACAAGATGCACCTTCTCTCCAGTGGTTG GGAGCTTTCTCACTCGATTCGTGCTGTTCGATGGAATAGTTTATGCAAATTGCAGGTCGATGTTGTGCAAGTGCAT GCAACCTGCTGCTGTGATCCTACAGCCGAGTTCATCCCCTGTAACTGTCATTGACATGTACAAATGTGTGAGTCTG CAGCTTGACAATCTCAGATTCACCATCACTCAATTGGCCAATGTAACCTACAATAGCACCATCAAGCTTGAATCAT CCCAGATCTTGTCTATTGATCCGTTGGATATATCCCAGAATCTAGCTGCGGTGAATAAGAGTCTAAGTGATGCACT ACAACACTTAGCACAAAGTGACACATATCTTTCTGCAATCACATCAGCTACGACTACAAGTGTATTATCCATAATA GCAATCTGTCTTGGATCGTTAGGTTTAATATTAATAATCTTGCTCAGTGTAGTTGTGTGGAAGTTATTGACCATTG TCGTTGCTAATCGAAATAGAATGGAGAATTTTGTTTATCATAAATAAGCATTCCACCACTCACGATCTGATCTCAG TGAGAAAAATCAACCTGCAACTCTTGGAACAAGATAAGACAGTCATCCATTAGTAATTTTTAAGAAAAAAACGATA GGACCGAACCTAGTATTGAAAGAACCGTCTCGGTCAATCTAGGTAATCGAGCTGATACCGTCTCGGAAAGCTCAAA TCATGCTGCCTGATCCGGAAGATCCGGAAAGCAAGAAAGCTACAAGGAGAGCAGGAAACCTAATTATCTGCTTCCT ATTCATCTTCTTTCTGTTTGTAACCTTCATTGTTCCAACTCTAAGACACTTGCTGTCCTAACACCTGCTATAGGCT ATCCACTGCATCATCTTACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGGT GCCAACAGCGCAATCCAGTCGACGCGGCCGCCACCATGAAATCAGGAGTGAAACTGGACGAGCACGCTAATCAGGG AGACGCAGTCAGCACCCAGCCAAACCCCGAGAATGTCGCACAGGTCACCGTGGACCCACTGAACGATCCCAATTCT CCTCTGGCCAAGCGGAGCGTGTACTTCGACTTTGATAGCTATTCCGTGCAGGACCAGTACCAGGCCCTGCTGCAGC AGCACGCCCAGTATCTGAAGAGCCACCCCCAGAGGCACATCCTGATCCAGGGCAACACCGATGAGCGCGGCACATC CGAGTACAATCTGGCCCTGGGCCAGAAGAGGGCAGAGGCCGTGCGGAGAGCCCTGTCCCTGCTGGGCGTGGGCGAC GCCCAGATGGAGGCCGTGTCTCTGGGCAAGGAGAAGCCCGTGGCCCTGGGGCACGACGAAGCCTCTTGGGCACAGA ATCGCCGAGCAGACCTGGTGTATCAGCAGTGATGAACGCGTGCTAGCTCTCCTGCCATACTTCCTACTCACATCAT ATCTATTTTAAAGAAAAAATAGGCCCGAACACTAATCGTGCCGGCAGTGCCACTGCACACACAACACTACACATAC AATACACTACAATGGTTGCAGAAGATGCCCCTGTTAGGGCCACTTGCCGAGTATTATTTCGAACAACAACTTTAAT CTTTCTATGCACACTACTAGCATTAAGCATCTCTATCCTTTATGAGAGTTTAATAACCCAAAAGCAAATCATGAGC CAAGCAGGCTCAACTGGATCTAATTCTGGATTAGGAAGTATCACTGATCTTCTTAATAATATTCTCTCTGTCGCAA ATCAGATTATATATAACTCTGCAGTCGCTCTACCTCTACAATTGGACACTCTTGAATCAACACTCCTTACAGCCAT TAAGTCTCTTCAAACCAGTGACAAGCTAGAACAGAACTGCTCGTGGAGTGCTGCACTGATTAATGATAATAGATAC ATTAATGGCATCAATCAGTTCTATTTTTCAATTGCTGAGGGTCGCAATCTGACACTTGGCCCACTTCTTAATATGC CTAGTTTCATTCCAACTGCCACGACACCAGAGGGCTGCACCAGGATCCCATCATTCTCGCTCACTAAGACACACTG GTGTTATACACACAATGTTATCCTGAATGGATGCCAGGATCATGTATCCTCAAATCAATTTGTTTCTATGGGAATC ATTGAACCCACTTCTGCCGGGTTTCCATTCTTTCGAACCCTAAAGACTCTATATCTCAGCGATGGGGTCAATCGTA AGAGCTGCTCTATCAGTACAGTTCCGGGGGGTTGTATGATGTACTGTTTTGTTTCTACTCAACCAGAGAGGGATGA CTACTTTTCTGCCGCTCCTCCAGAACAACGAATTATTATAATGTACTATAATGATACAATCGTGGAGCGCATAATT AATCCACCCGGGGTACTAGATGTATGGGCAACATTGAACCCAGGAACAGGAAGCGGGGTATATTATTTAGGTTGGG TGCTCTTTCCAATATATGGCGGCGTGATTAAAGGTACGAGTTTATGGAATAATCAAGCAAATAAATACTTTATCCC CCAGATGGTTGCTGCTCTCTGCTCACAAAACCAGGCAACTCAAGTCCAAAATGCTAAGTCATCATACTATAGCAGC TGGTTTGGCAATCGAATGATTCAGTCTGGGATCCTGGCATGTCCTCTTCGACAGGATCTAACCAATGAGTGTTTAG TTCTGCCCTTTTCTAATGATCAGGTGCTTATGGGTGCTGAAGGGAGATTATACATGTATGGTGACTCGGTGTATTA CTATCAAAGAAGCAATAGTTGGTGGCCTATGACCATGCTGTATAAGGTAACCATAACATTCACTAATGGTCAGCCA TCTGCTATATCAGCTCAGAATGTGCCCACACAGCAGGTCCCTAGACCTGGGACAGGAGACTGCTCTGCAACCAATA GATGTCCCGGTTTTTGCTTGACAGGAGTGTATGCCGATGCCTGGTTACTGACCAACCCTTCGTCTACCAGTACATT TGGATCAGAAGCAACCTTCACTGGTTCTTATCTCAACACAGCAACTCAGCGTATCAATCCGACGATGTATATCGCG AACAACACACAGATCATAAGCTCACAGCAATTTGGATCAAGCGGTCAAGAAGCAGCATATGGCCACACAACTTGTT TTAGGGACACAGGCTCTGTTATGGTATACTGTATCTATATTATTGAATTGTCCTCATCTCTCTTAGGACAATTTCA GATTGTCCCATTTATCCGTCAGGTGACACTATCCTAAAGGCAGAAGCCTTCAGGTCTGACCCAGCCAATCAAAGCA TTATACCAGACCATGGAATGCATACCAAACATTATTGACACTAATGACACACAAAATTGGTTTTAAGAAAAACCAA GAGAACAATAGGCCAGAATGGCTGGGTCTCGGGAGATATTACTCCCTGAAGTCCATCTCAATTCACCAATTGTAAA GCATAAGCTATACTATTACATTCTACTTGGAAACCTCCCAAATGAGATCGACCTTGACGATTTAGGTCCATTACAT AATCAAAATTGGAATCAGATAGCACATGAAGAGTCTAACTTAGCTCAACGCTTGGTAAATGTAAGAAATTTTCTAA TTACCCACATCCCTGATCTTAGAAAGGGCCATTGGCAAGAGTATGTCAATGTAATACTGTGGCCGCGAATTCTTCC CTTGATCCCGGATTTTAAAATCAATGACCAATTGCCTCTGCTCAAAAATTGGGACAAGTTAGTTAAAGAATCATGT TCAGTAATCAATGCAGGTACTTCCCAGTGCATTCAGAATCTCAGCTATGGACTGACAGGTCGTGGGAACCTCTTTA CACGATCACGTGAACTCTCTGGTGACCGCAGGGATATTGATCTTAAGACAGTTGTGGCAGCATGGCATGACTCAGA CTGGAAAAGAATAAGTGATTTTTGGATTATGATCAAATTCCAGATGAGACAATTAATTGTTAGGCAAACAGATCAT AATGATTCTGATTTAATCACGTATATCGAAAATAGAGAAGGCATAATCATCATAACCCCTGAACTGGTAGCATTAT TTAACACTGAGAATCATACACTAACATACATGACCTTTGAAATTGTACTGATGGTTTCAGATATGTACGAAGGTCG TCACAACATTTTATCACTATGCACAGTTAGCACTTACCTGAATCCTCTGAAGAAAAGAATAACATATTTATTGAGC CTTGTAGATAACTTAGCTTTTCAGATAGGTGATGCTGTATATAACATAATTGCTTTGCTAGAATCCTTTGTATATG CACAGTTGCAAATGTCAGATCCCATCCCAGAACTCAGAGGACAATTCCATGCATTCGTATGTTCTGAGATTCTTGA TGCACTAAGAGGAACTAATAGTTTCACCCAGGATGAATTAAGAACTGTGACAACTAATTTGATATCCCCATTCCAA GATCTGACCCCAGATCTTACGGCTGAATTGCTCTGTATAATGAGGCTTTGGGGACACCCCATGCTCACTGCCAGTC AAGCTGCAGGAAAGGTACGCGAGTCTATGTGTGCTGGAAAAGTATTAGACTTTCCCACCATTATGAAAACACTAGC CTTTTTCCATACTATTCTGATCAATGGATACAGGAGGAAGCATCATGGAGTATGGCCACCCTTAAACTTACCGGGT AATGCTTCAAAGGGTCTCACGGAACTTATGAATGACAATACTGAAATAAGCTATGAATTCACACTTAAGCATTGGA AGGAAGTCTCTCTTATAAAATTCAAGAAATGTTTTGATGCAGACGCAGGTGAGGAACTCAGTATATTTATGAAAGA TAAGGCAATTAGTGCCCCAAAACAAGACTGGATGAGTGTGTTTAGAAGAAGCCTAATCAAACAGCGCCATCAGCAT CATCAGGTCCCCCTACCAAATCCATTCAATCGACGGCTGTTGCTAAACTTTCTCGGAGATGACAAATTCGACCCGA ATGTGGAGCTACAGTATGTAACATCAGGTGAGTATCTACATGATGACACGTTTTGTGCATCATATTCACTAAAAGA GAAGGAAATTAAACCTGATGGTCGAATTTTTGCAAAGTTGACTAAGAGAATGAGATCATGTCAAGTTATAGCAGAA TCCTTTTTAGGAACCATGCTGGGAAGTTAATGAAAGAGAATGGTGTTGTGATGAATCAGCTATCATTAACAAAAT CACTATTAACAATGAGTCAGATTGGAATAATATCCGAGAAAGCTAGAAAGTCAACTCGAGATAACATAAATCAACC TGGTTTCCAGAATATCCAGAGAAATAAATCACATCACTCCAAGCAAGTCAATCAGCGAGATCCAAGTGATGACTTT GAATTGGCAGCATCTTTTTTAACTACTGATCTCAAAAAATATTGTTTACAATGGAGGTACCAGACAATTATCCCAT TTGCTCAATCATTAAACAGAATGTATGGTTATTCCTCATCTCTTTGAGTGGATTCACTTACGGCTAATGCGTAGTAC ACTTTACGTGGGGGATCCCTTCAACCCACCAGCAGATACCAGTCAATTTGATCTAGATAAAGTAATTAATGGAGAT ATCTTCATTGTATCACCCAGAGGTGGAATTGAAGGGCTGTGTCAAAAGGCTTGGACAATGATATCTATCGCTGTGA TAATTCTATCTGCCACAGAGTCTGACACGAGTAATGAGTATGGTGCAGGGAGATAATCAAGCAATTGCTGTCAC CACACGAGTACCAAGGAGCCTGCCGACTCTTGAGAAAAAGACTATTGCTTTAGATCTTGTAATCTATTCTTTTGAG AGGTTAAAATGTAATAATTTTGGATTAGGTCACCATTTGAAAGAACAAGAGACTATCATTAGTTCTCACTTCTTTG TTTATAGCAAGAGAATATTCTATCAGGGGAGGATTCTAACGCAAGCCTTAAAAAATGCTAGTAAGCTCTGCTTGAC AGTCTGATGTCCTAGGAGAATGCACCCAATCATCATGTTCTAATCTTGCAACTACTGTCATGAGGTTAACTGAGAAT GGTGTTGAAAAAGATATCTGTTTCTACTTGAATATCTATATGACCATCAAACAGCTCTCCTATGATATCATCTTCC CTCAAGTGTCAATTCCTGGAGATCAGATCACATTAGAATACATAAATAATCCACACCTGGTATCACGATTGGCTCT TTTGCCATCCCAGTTAGGAGGTCTAAACTACCTGTCATGCAGTAGGCTGTTCAATCGAAACATAGGCGACCCGGTG GTTTCCGCAGTTGCAGATCTTAAGAGATTAATTAAATCAGGATGTATGGATTACTGGATCCTTTATAACTTATTAG GGAGAAAACCGGGAAACGGCTCATGGGCTACTTTAGCAGCTGACCCGTACTCAATCAATATAGAGTATCAATACCC TCCAACTACAGCTCTTAAGAGGCACACCCAACAAGCTCTGATGGAACTCAGTACGAATCCAATGTTACGTGGCATA TTCTCTGACAATGCACAGGCAGAAGAAAATAACCTTGCTAGGTTTCTCCTGGATAGGGAGGTGATCTTTCCGCGTG TAGCTCACATCATCATTGAGCAAACCAGTGTCGGGAGGAGAAAACAGATTCAAGGATATTTGGATTCAACTAGATC GATAATGAGGAAATCACTAGAAATTAAGCCCTTATCCAATAGGAAGCTTAATGAAATACTGGATTACAACATCAAT TACCTAGCTTACAATTTGGCATTACTCAAGAATGCTATTGAACCTCCGACTTATTTGAAGGCAATGACACTTGAAA CATGTAGCATCGACATTGCAAGGAACCTCCGGAAGCTCTCCTGGGCCCCACTCTTGGGTGGGAGAAATCTTGAAGG ATTAGAGACGCCAGATCCCATTGAAATTACTGCAGGAGCATTAATTGTTGGATCGGGCTACTGTGAACAGTGTGCT GCAGGAGACAATCGATTCACATGGTTTTTCTTGCCATCTGGTATCGAGATAGGAGGGGATCCCCGTGATAATCCTC CTATCCGTGTACCGTACATTGGCTCCAGGACTGATGAGAGGAGGGTAGCCTCAATGGCATACATCAGGGGTGCCTC GAGTAGCTTAAAAGCAGTTCTTAGACTGGCGGGAGTGTACATCTGGGCATTCGGAGATACTCTGGAGAATTGGATA GATGCACTGGATTTGTCTCACACTAGAGTTAACATCACACTTGAACAGCTGCAATCCCTCACCCCACTTCCAACCT CTGCCAATCTAACCCATCGGTTGGATGATGGCACAACTACCCTAAAGTTTACTCCTGCGAGCTCTTACACCTTTTC AAGTTTCACTCATATATCAAATGATGAGCAATACCTGACAATTAATGACAAAACTGCAGATTCAAATATAATCTAC CAACAGTTAATGATCACTGGACTCGGAATCTTAGAAACATGGAATAATCCCCCAATCAATAGAACATTCGAAGAAT CTACCCTACATTTGCACACTGGTGCATCATGTTGTGTCCGACCTGTGGACTCCTGCATTCTCTCAGAAGCATTAAC AGTCAAGCCACATATTACAGTACCGTACAGCAATAAATTTGTATTTGATGAGGACCCGCTATCTGAATATGAAACT GCAAAACTGGAATCGTTATCATTCCAAGCCCAATTAGGCAACATTGATGCTGTAGATATGACAGGTAAATTAACAT TATTGTCCCAATTCACTGCAAGGCAGATTATCAATGCAATCACTGGACTCGATGAGTCTGTCTCTCTTACTAATGA TGCCATTGTTGCATCAGACTATGTCTCCAATTGGATTAGTGAATGCATGTATACCAAATTAGATGAATTATTTATG TATTGTGGGTGGGAACTACTATTGGAACTATCCTATCAAATGTATTATCTGAGGGTAGTTGGGTGGAGTAATATAG TGGATTATTCTTACATGATCTTGAGAAGAATCCCGGGTGCAGCATTAAACAATCTGGCATCTACATTAAGTCATCC AAAACTTTTCCGACGAGCTATCAACCTAGATATAGTTGCCCCCTTAAATGCTCCTCATTTTGCATCTCTGGACTAC ATCAAGATGAGTGTGGATGCAATACTCTGGGGCTGTAAAAGAGTCATCAATGTGCTCTCCAATGGAGGGGACTTAG AATTAGTTGTGACATCTGAAGATAGCCTTATTCTCAGTGACCGATCCATGAATCTCATTGCAAGGAAATTAACTTT ATTATCACTGATTCACCATAATGGTTTGGAACTACCAAAGATTAAGGGGTTCTCTCCTGATGAGAAGTGTTTCGCT TTGACAGAATTTTTGAGGAAAGTGGTGAACTCAGGGTTGAGTTCAATAGAGAACCTATCAAATTTTATGTACAATG TGGAGAACCCACGGCTTGCAGCATTCGCCAGCAACAATTACTACCTGACCAGAAAATTATTGAATTCAATACGAGA TACTGAGTCGGGTCAAGTAGCAGTCACCTCATATTATGAATCATTAGAATATATTGATAGTCTTAAGCTAACCCCA CATGTGCCTGGCACCTCATGCATTGAGGATGATAGTCTATGTACAAATGATTACATAATCTGGATCATAGAGTCTA ATGCAAACTTGGAGAAGTATCCAATTCCAAATAGCCCTGAGGATGATTCCAATTTCCATAACTTTAAGTTGAATGC TCCATCGCACCATACCTTACGCCCATTAGGGTTGTCATCAACTGCTTGGTATAAGGGTATAAGCTGCTGCAGGTAC CTTGAGCGATTAAAGCTACCACAAGGTGATCATTTATATATTGCAGAAGGTAGTGGTGCCAGTATGACAATCATAG AATACCTATTCCCAGGAAGAAAGATATATTACAATTCTTTATTTAGTAGTGGTGACAATCCCCCACAAAGAAATTA TGCACCAATGCCTACTCAGTTCATTGAGAGTGTCCCATACAAGCTCTGGCAAGCACACACAGATCAATATCCCGAG ATTTTTGAGGACTTCATCCCTCTATGGAACGGAAACGCCGCCATGACTGACATAGGAATGACAGCTTGTGTAGAAT TCATCATCAATCGAGTCGGCCCAAGGACTTGCAGTTTAGTACATGTAGATTTGGAATCAAGTGCAAGCTTAAATCA ACAATGCCTGTCAAAGCCGATAATTAATGCTATCATCACTGCTACAACTGTTTTGTGCCCTCATGGGGTGCTTATT CTGAAATATAGTTGGTTGCCATTTACTAGATTTAGTACTTTGATCACTTTCTTATGGTGCTACTTTGAGAGAATCA CTGTTCTTAGGAGCACATATTCTGATCCAGCTAATCATGAGGTTTATTTAATTTGTATCCTTGCCAACAACTTTGC ATTCCAGACTGTCTCGCAGGCAACAGGAATGGCGATGACTTTAACTGATCAAGGGTTTACTTTGATATCACCTGAA AGAATAAATCAGTATTGGGATGGTCACTTGAAGCAAGAACGTATCGTAGCAGAAGCAATTGATAAGGTGGTTCTAG GAGAAAATGCTCTATTTAATTCGAGTGATAATGAATTAATTCTCAAATGTGGAGGGACACCAAATGCACGGAATCT CATCGATATCGAGCCAGTCGCAACTTTCATAGAATTTGAACAATTGATCTGCACAATGTTGACAACCCACTTGAAG GAAATAATTGATATAACAAGGTCTGGAACCCAGGATTATGAAAGTTTATTACTCACTCCTTACAATTTAGGTCTTC TTGGTAAAATCAGTACGATAGTGAGATTATTAACAGAAAGGATTCTAAATCATACTATCAGGAATTGGTTGATCCT CCCACCTTCGCTCCGGATGATCGTGAAGCAGGACTTGGAATTCGGCATATTCAGGATTACTTCCATCCTCAATTCT GATCGGTTCCTGAAGCTTTCTCCAAATAGGAAATACTTGATTGCACATTAACTGCAGGCTACATTAGGAAATTGA TTGAGGGGGATTGCAATATCGATCTAACCAGACCTATCCAAGCAAATCTGGAAAGCATTAGGTTGTGTAGTCTA TTGTCACGATCCAATGGATCAAAGGGAGTCAACAGAGTTTATTGATATAAATATTAATGAAGAAATAGACCGCGGG ATCGATGGCGAGGAAATCTAAACATATCAAGAATCAGAATTAGTTTAAGAAAAAAGAAGAGGATTAATCTTGGTTT TCCCCTTGGT

[0110] The underlined sequence is the PIV5 genomic nucleic acid sequence. The italicized and underlined sequences are the entire Pal nucleic acid sequence.

[0111] b. pMHD39-PIV5-NTTM-Pal

[0112] The pMHD39-PIV5-NTTM-Pal nucleic acid sequence provided in this article, from 5' to 3', is as follows:

[0113] ACCAAGGGGAAAATGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCTA TGGCCTTCGTGACCGACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCGACA CAAAAATGTCATCCGTGCTTAAAGCATATGAGCGATTCACGCTCACTCAAGAACTGCAAGATCAGAGTGAGGAAGGT ACAATCCCACCTACAACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGATCCCG GCTTCTTCTATTCTGCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACTCACAA TGTTTTCGCTACCATCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATCGAAAGG GTAGAGATCGATGGCTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCTCGTTCAGGTATGAGCCGTGGAGAGAT CAATGCCTATGCTGCACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTCGTTGATTCCGAAGTCG AGGGAACTGCATGGGATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGGATAGTGACT TGCAAGTGCATGACTGCGCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAATATCGTCAGCAAGGCAG GATCAACCCGAGATATCTCCTGCAACCGGAGGCTCGACGAATAATCCAGAATGTAATCCGGAAGGGAATGGTGGTCA GACATTTCCTCACCTTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTATGCTATGGTAGGG GATGTTGGAAAGTATATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAATATGCATTAGGAACTAGATG GCCCACACTTGCTTTAGCTGCATTTTCAGGAGAGCTAACAAAGCTAAAGTCCCTCATGGCATTATACCAGACCCTTG GTGAGCAGGCCCGATATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTTGCTGCAGCAAACTACCCACTGCTA TATAGCTATGCTATGGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTCTCCAGATCATACATGAA CAAGACATATTTCCAATTGGGAATGGAAACTGCAAGAAAACAACAGGGTGCAGTTGACATGAGGATGGCAGAAGATC TCGGTCTAACTCAAGCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACAGCAAATCGAGGGGCAGAC ACCAGGGGAGGAGTCAACCCGTTCTCATCTGTCACTGGGACAACTCAGGTGCCCGCTGCAGCAACAGGTGACACACT CGAGAGTTACATGGCAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCATGATGATGAGATGCCACCAT TGGAAGAGGAGGAAGAGGACGACACATCTGCAGGTCCACGCACTGGACCAACTCTTGAACAAGTGGCCTTGGACATC CAGAACGCAGCAGTTGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGGGTGATCTTGACATCTAGACAAT TCAGATCCCAATCTAAAATTGACATACCTAATTGATTAGTTAGATGGAACTACAGTGGATTCCATAAGGTTCCTGCC TACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGGTGCCAACAGCGCAATCCA CAATCTACAATGGATCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATAAGCTCATAGAGACAGGCCTGAATAC TGTAGAGTATTTTACTTCCCAACAAGTCACAGGAACATCCTCTCTTGGAAAGAATACAATACCACCAGGGGTCACAG GACTACTAACCAATGCTGCAGAGGCAAAGATCCAAGAGTCAACTAACCATCAGAAGGGCTCAGTTGGTGGGGGTGCA AAACCAAAGAAACCGCGACCAAAAATTGCCATTGTGCCAGCAGATGACAAAACAGTGCCCGGAAAGCCGATCCCAAA CCCTCTATTAGGTCTGGACTCCACCCCGAGCACCCAAACTGTGCTTGATCTAAGTGGGAAAACATTACCATCAGGAT CCTATAAGGGGGTTAAGCTTGCGAAATTTGGAAAAGAAAATCTGATGACACGGTTCATCGAGGAACCCAGAGAGAAT CCTATCGCAACCAGTTCCCCCATCGATTTTAAGAGGGGCAGGGATACCGGCGGGTTCCATAGAAGGGAGTACTCAAT CGGATGGGTGGGAGATGAAGTCAAGGTCACTGAGTGGTGCAATCCATCCTGTTCTCCAATCACCGCTGCAGCAAGGC GATTTGAATGCACTTGTCACCAGTGTCCAGTCACTTGCTCTGAATGTGAACGAGATACTTAATACAGTGAGAAATTT GGACTCTCGGATGAATCAACTGGAGACAAAAGTAGATCGCATTCTCTCATCTCAGTCTCTAATCCAGACCATCAAGA ATGACATAGTTGGACTTAAAGCAGGGATGGCTACTTTAGAAGGAATGATTACAACTGTGAAAATCATGGACCCGGGA GTTCCCAGTAATGTTACTGTGGAAGATGTACGCAAGACACTAAGTAACCATGCTGTTGTTGTGCCAGAATCATTCAA TGATAGTTTCTTGACTCAATCTGAAGATGTAATTTCACTTGATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAGA AGATTGTCAGGAAGGTTCCTCCTCAGAAGGATCTGACTGGATTGAAGATTACACTAGAGCAATTGGCAAAGGATTGC ATCAGCAAACCGAAGATGAGGGAAGAGTATCTCCTCAAAATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCT CAAGAAAGCAATCATCCGCAGTGCAATTTGATCAAGAAACACCCAATTACACTACACTGGTATGACACTGTACTAAC CCTGAGGGTTTTAGAAAAAACGATTAACGATAAATAAGCCCGAACACTACACACTACCTGAGGCAGCCATGCCATCC ATCAGCATTCCCGCAGACCCCACCAATCCACGTCAATCAATAAAAGCGTTCCCAATTGTGATCAACAGTGATGGGGG TGAGAAAGGCCGCTTGGTTAAACAACTACGCACAACCTACTTGAATGACCTAGATACTCATGAGCCACTGGTGACAT TCATAAATACCTATGGATTCATCTACGAACAGGATCGGGGGAATACCATTGTCGGAGAGGATCAACTTGGGAAGAAA AGAGAGGCTGTGACCGCTGCAATGGTTACCCTTGGATGTGGGCCTAATCTACCATCATTAGGGAATGTCCTGGGACA ACTGAGGGAATTCCAGGTCACTGTTAGGAAGACATCCAGCAAAGCGGAAGAGATGGTCTTTGAAATTGTTAAGTATC CGAGAATATTTCGGGGTCATACATTAATCCAGAAAGGACTAGTCTGTGTCTCCGCAGAAAAATTTGTTAAGTCACCA GGGAAAATACAATCTGGAATGGACTATCTCTTCATTCCGACATTTCTGTCAGTGACTTACTGTCCAGCTGCAATCAA ATTTCAGGTACCTGGCCCCATGTTGAAAATGAGATCAAGATACACTCAGAGCTTACAACTTGAACTAATGATAAGAA TCCTGTGTAAGCCCGATTCGCCACTTATGAAGGTCCATACCCCTGACAAGGAGGGAAGAGGATGTCTTGTATCAGTA TGGCTGCATGTATGCAACATCTTCAAATCAGGAAACAAGAATGGCAGTGAGTGGCAGGAATACTGGATGAGAAAGTG TGCTAACATGCAACTTGAAGTGTCGATTGCAGATATGTGGGGACCAACTATCATAATTCATGCCAGAGGTCACATTC CCAAAAGTGCTAAGTTGTTTTTTGGAAAGGGTGGATGGAGCTGCCATCCACTTCACGAAGTTGTTCCAAGTGTCACT AAAACACTATGGTCCGTGGGCTGTGAGATTACAAAGGCGAAGGCAATAATACAAGAGAGTAGCATCTCTCTTCTCGT GGAGACTACTGACATCATAAGTCCAAAAGTCAAAATTTCATCTAAGCATCGCCGCTTTGGGAAATCAAATTGGGGTC TGTTCAAGAAAACTAAATCACTGCCTAACCTGACGGAGCTGGAATGACTGACCTCTAATCGAGACTACACCGCCGCA AACTATAGGTGGGTGGTACCTCAGTGATTAATCTTGTAAGCACTGATCGTAGGCTACAACACACTAATATTATCCAG ATTAGAGAGCTTAATTAGCTCTGTATTAATAATAACACTACTATTCCAATAACTGGAATCACCAGCTTGATTTATCT CCAAAATGATTCAAAGAAAACAAATCATATTAAGACTATCCTAAGCACGAACCCATATCGTCCTTCAAATCATGGGT ACTATAATTCAATTTCTGGTGGTCTCCTGTCTATTGGCAGGAGCAGGCAGCCTTGATCCAGCAGCCCTCATGCAAAT CGGTGTCATTCCAACAAATGTCCGGCAACTTATGTATTATACTGAAGCTTCATCAGCATTCATTGTTGTGAAGTTAA TGCCTACAATTGACTCGCCGATTAGTGGATGTAATATAACATCAATTTCAAGCTATAATGCAACAGTGACAAAACTC CTACAGCCGATCGGTGAGAATTTGGAGACGATTAGGAACCAGTTGATTCCAACTCGGAGGAGACGCCGGTTTGCAGG GGTGGTGATTGGATTAGCTGCATTAGGAGTAGCTACTGCCGCACAGGTCACTGCCGCAGTGGCACTAGTAAAGGCAA ATGAAAATGCTGCGGCTATACTCAATCTCAAAAATGCAATCCAAAAAACAAATGCAGCAGTTGCAGATGTGGTCCAG GCCACACAATCACTAGGAACGGCAGTTCAAGCAGTTCAAGATCACATAAACAGTGTGGTAAGTCCAGCAATTACAGC AGCCAATTGTAAGGCCCAAGATGCTATCATTGGCTCAATCCTCAATCTCTATTTGACCGAGTTGACAACCATCTTCC ACAATCAAATTACAAACCCTGCATTGAGTCCCATTACAATTCAAGCTTTAAGGATCCTACTGGGGAGTACCTTGCCG ACTGTGGTCGAAAAATCTTTCAATACCCAGATAAGTGCAGCTGAGCTTCTCTCATCAGGGTTATTGACAGGCCAGAT TGTGGGATTAGATTTGACCTATATGCAGATGGTCATAAAAATTGAGCTGCCAACTTTAACTGTACAACCTGCAACCC AGATCATAGATCTGGCCACCATTTCTGCATTCATTAACAATCAAGAAGTCATGGCCCAATTACCAACACGTGTTATG GTGACTGGCAGCTTGATCCAAGCCTATCCCGCATCGCAATGCACCATTACACCCAACACTGTGTACTGTAGGTATAA TGATGCCCAAGTACTCTCAGATGATACTATGGCTTGCCTCCAAGGTAACTTGACAAGATGCACCTTCTCTCCAGTGG TTGGGAGCTTTCTCACTCGATTCGTGCTGTTCGATGGAATAGTTTATGCAAATTGCAGGTCGATGTTGTGCAAGTGC ATGCAACCTGCTGCTGTGATCCTACAGCCGAGTTCATCCCCTGTAACTGTCATTGACATGTACAAATGTGTGAGTCT GCAGCTTGACAATTCTCAGATTCACCATCACTCAATTGGCCAATGTAACCTACAATAGCACCATCAAGCTTGAATCAT CCCAGATCTTGTCTATTGATCCGTTGGATATATCCCAGAATCTAGCTGCGGTGAATAAGAGTCTAAGTGATGCACTA CAACACTTAGCACAAAGTGACACATATCTTTCTGCAATCACATCAGCTACGACTACAAGTGTATTATCCATAATAGC AATCTGTCTTGGATCGTTAGGTTTAATATTAATAATCTTGCTCAGTGTAGTTGTGTGGAAGTTATTGACCATTGTCG TTGCTAATCGAAATAGAATGGAGAATTTTGTTTATCATAAATAAGCATTCCACCACTCACGATCTGATCTCAGTGAG AAAAATCAACCTGCAACTCTTGGAACAAGATAAGACAGTCATCCATTAGTAATTTTTAAGAAAAAACGATAGGACC GAACCTAGTATTGAAAGAACCGTCTCGGTCAATCTAGGTAATCGAGCTGATACCGTCTCGGAAAGCTCAAATCATGC TGCCTGATCCGGAAGATCCGGAAAGCAAGAAAGCTACAAGGAGAGCAGGAAACCTAATTATCTGCTTCCTATTCATC TTCTTTCTGTTTGTAACCTTCATTGTTCCAACTCTAAGACACTTGCTGTCCTAACACCTGCTATAGGCTATCCACTG CATCATCTTACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGAC

[0114]

[0115] AGCACGCTAATCAGGGAGACGCAGTCAGCACCCAGCCAAACCCCGAGAATGTCGCACAGGTCACCGTG GACCCACTGAACGATCCCAATTCTCCTCTGGCCAAGCGGAGCGTGTACTTCGACTTTGATAGCTATTCCGTGCAGG ACCAGTACCAGGCCCTGCTGCAGCAGCACGCCCAGTATCTGAAGAGCCACCCCCAGAGGCACATCCTGATCCAGGG CAACACCGATGAGCGCGGCACATCCGAGTACAATCTGGCCCTGGGCCAGAAGAGGGCAGAGGCCGTGCGGAGAGCC CTGTCCCTGCTGGGCGTGGGCGACGCCCAGATGGAGGCCGTGTCTCTGGGCAAGGAGAAGCCCGTGGCCCTGGGGC ACGACGAAGCCTCTTGGGCACAGAATCGCCGAGCAGACCTGGTGTATCAGCAGTGATGAACGCGTGCTAGCTCTCC TGCCATACTTCCTACTCACATCATATCTATTTTAAAGAAAAAATAGGCCCGAACACTAATCGTGCCGGCAGTGCCA CTGCACACACAACACTACACATACAATACACTACAATGGTTGCAGAAGATGCCCCTGTTAGGGCCACTTGCCGAGT ATTATTTCGAACAACAACTTTAATCTTTCTATGCACACTACTAGCATTAAGCATCTCTATCCTTTATGAGAGTTTA ATAACCCAAAAGCAAATCATGAGCCAAGCAGGCTCAACTGGATCTAATTCTGGATTAGGAAGTATCACTGATCTTC TTAATAATATTCTCTCTGTCGCAAATCAGATTATATATAACTCTGCAGTCGCTCTACCTCTACAATTGGACACTCT TGAATCAACACTCCTTACAGCCATTAAGTCTCTTCAAACCAGTGACAAGCTAGAACAGAACTGCTCGTGGAGTGCT GCACTGATTAATGATAATAGATACATTAATGGCATCAATCAGTTCTATTTTTCAATTGCTGAGGGTCGCAATCTGA CACTTGGCCCACTTCTTAATATGCCTAGTTTCATTCCAACTGCCACGACACCAGAGGGCTGCACCAGGATCCCATC ATTCTCGCTCACTAAGACACACTGGTGTTATACACACAATGTTATCCTGAATGGATGCCAGGATCATGTATCCTCA AATCAATTTGTTTCTATGGGAATCATTGAACCCACTTCTGCCGGGTTTCCATTCTTTCGAACCCTAAAGACTCTAT ATCTCAGCGATGGGGTCAATCGTAAGAGCTGCTCTATCAGTACAGTTCCGGGGGGTTGTATGATGTACTGTTTTGT TTCTACTCAACCAGAGAGGGATGACTACTTTTCTGCCGCTCCTCCAGAACAACGAATTATTATAATGTACTATAAT GATACAATCGTGGAGCGCATAATTAATCCACCCGGGGTACTAGATGTATGGGCAACATTGAACCCAGGAACAGGAA GCGGGGTATATTATTTAGGTTGGGTGCTCTTTCCAATATATGGCGGCGTGATTAAAGGTACGAGTTTATGGAATAA TCAAGCAAATAAATACTTTATCCCCCAGATGGTTGCTGCTCTCTGCTCACAAAACCAGGCAACTCAAGTCCAAAAT GCTAAGTCATCATACTATAGCAGCTGGTTTGGCAATCGAATGATTCAGTCTGGGATCCTGGCATGTCCTCTTCGAC AGGATCTAACCAATGAGTGTTTAGTTCTGCCCTTTTCTAATGATCAGGTGCTTATGGGTGCTGAAGGGAGATTATA CATGTATGGTGACTCGGTGTATTACTATCAAAGAAGCAATAGTTGGTGGCCTATGACCATGCTGTATAAGGTAACC ATAACATTCACTAATGGTCAGCCATCTGCTATATCAGCTCAGAATGTGCCCACACAGCAGGTCCCTAGACCTGGGA CAGGAGACTGCTCTGCAACCAATAGATGTCCCGGTTTTTGCTTGACAGGAGTGTATGCCGATGCCTGGTTACTGAC CAACCCTTCGTCTACCAGTACATTTGGATCAGAAGCAACCTTCACTGGTTCTTATCTCAACACAGCAACTCAGCGT ATCAATCCGACGATGTATATCGCGAACAACACACAGATCATAAGCTCACAGCAATTTGGATCAAGCGGTCAAGAAG CAGCATATGGCCACACAACTTGTTTTAGGGACACAGGCTCTGTTATGGTATACTGTATCTATATTATTGAATTGTC CTCATCTCTCTTAGGACAATTTCAGATTGTCCCATTTATCCGTCAGGTGACACTATCCTAAAGGCAGAAGCCTTCA GGTCTGACCCAGCCAATCAAAGCATTATACCAGACCATGGAATGCATACCAAACATTATTGACACTAATGACACAC AAAATTGGTTTTAAGAAAAACCAAGAGAACAATAGGCCAGAATGGCTGGGTCTCGGGAGATATTACTCCCTGAAGT CCATCTCAATTCACCAATTGTAAAGCATAAGCTATACTATTACATTCTACTTGGAAACCTCCCAAATGAGATCGAC CTTGACGATTTAGGTCCATTACATAATCAAAATTGGAATCAGATAGCACATGAAGAGTCTAACTTAGCTCAACGCT TGGTAAATGTAAGAAATTTTCTAATTACCCACATCCCTGATCTTAGAAAGGGCCATTGGCAAGAGTATGTCAATGT AATACTGTGGCCGCGAATTCTTCCCTTGATCCCGGATTTTAAAATCAATGACCAATTGCCTCTGCTCAAAAATTGG GACAAGTTAGTTAAAGAATCATGTTCAGTAATCAATGCAGGTACTTCCCAGTGCATTCAGAATCTCAGCTATGGAC TGACAGGTCGTGGGAACCTCTTTACACGATCACGTGAACTCTCTGGTGACCGCAGGGATATTGATCTTAAGACAGT TGTGGCAGCATGGCATGACTCAGACTGGAAAAGAATAAGTGATTTTTGGATTATGATCAAATTCCAGATGAGACAA TTAATTGTTAGGCAAACAGATCATAATGATTCTGATTTAATCACGTATATCGAAAATAGAGAAGGCATAATCATCA TAACCCCTGAACTGGTAGCATTATTTAACACTGAGAATCATACACTAACATACATGACCTTTGAAATTGTACTGAT GGTTTCAGATATGTACGAAGGTCGTCACAACATTTTATCACTATGCACAGTTAGCACTTACCTGAATCCTCTGAAG AAAAGAATAACATATTTATTGAGCCTTGTAGATAACTTAGCTTTTCAGATAGGTGATGCTGTATATAACATAATTG CTTTGCTAGAATCCTTTGTATATGCACAGTTGCAAATGTCAGATCCCATCCCAGAACTCAGAGGACAATTCCATGC ATTCGTATGTTCTGAGATTCTTGATGCACTAAGAGGAACTAATAGTTTCACCCAGGATGAATTAAGAACTGTGACA ACTAATTTGATATCCCCATTCCAAGATCTGACCCCAGATCTTACGGCTGAATTGCTCTGTATAATGAGGCTTTGGG GACACCCCATGCTCACTGCCAGTCAAGCTGCAGGAAAGGTACGCGAGTCTATGTGTGCTGGAAAAGTATTAGACTT TCCCACCATTATGAAAACACTAGCCTTTTTCCATACTATTCTGATCAATGGATACAGGAGGAAGCATCATGGAGTA TGGCCACCCTTAAACTTACCGGGTAATGCTTCAAAGGGTCTCACGGAACTTATGAATGACAATACTGAAATAAGCT ATGAATTCACACTTAAGCATTGGAAGGAAGTCTCTCTTATAAAATTCAAGAAATGTTTTGATGCAGACGCAGGTGA GGAACTCAGTATATTTATGAAAGATAAGGCAATTAGTGCCCCAAAACAAGACTGGATGAGTTGTTTAGAAGAAGC CTAATCAAACAGCGCCATCAGCATCATCAGGTCCCCCTACCAAATCCATTCAATCGACGGCTGTTGCTAAACTTTC TCGGAGATGACAAATTCGACCCGAATGTGGAGCTACAGTATGTAACATCAGGTGAGTATCTACATGATGACACGTT TTGTGCATCATATTCACTAAAAGAGAAGGAAATTAAACCTGATGGTCGAATTTTTGCAAAGTTGACTAAGAGAATG AGATCATGTCAAGTTATAGCAGAATCTCTTTTAGCGAACCATGCTGGGAAGTTAATGAAAGAGAATGGTGTTGTGA TGAATCAGCTATCATTAACAAAATCACTATTAACAATGAGTCAGATTGGAATAATATCCGAGAAAGCTAGAAAGTC AACTCGAGATAACATAAATCAACCTGGTTTCCAGAATATCCAGAGAAATAAATCACATCACTCCAAGCAAGTCAAT CAGCGAGATCCAAGTGATGACTTTGAATTGGCAGCATCTTTTTTAACTACTGATCTCAAAAAATATTGTTTACAAT GGAGGTACCAGACAATTATCCCATTTGCTCAATCATTAAACAGAATGTATGGTTATTCCTCATCTCTTTGAGTGGAT TCACTTACGGCTAATGCGTAGTACACTTTACGTGGGGGATCCCTTCAACCCACCAGCAGATACCAGTCAATTTGAT CTAGATAAAGTAATTAATGGAGATATCTTCATTGTATCACCCAGAGGTGGAATTGAAGGGCTGTGTCAAAAGGCTT GGACAATGATATCTATCGCTGTGATAATTCTATCTGCCACAGAGTCTGGCACACGAGTAATGAGTATGGTGCAGGG AGATAATCAAGCAATTGCTGTCACCACAGAGTACCAAGGAGCCTGCCGACTCTTGAGAAAAAGACTATTGCTTTT AGATCTTGTAATCTATTCTTTGAGAGGTTAAAATGTAATAATTTTGGATTAGGTCACCATTTGAAAGAACAAGAGA CTATCATTAGTTCTCACTTCTTTGTTTATAGCAAGAGAATATTCTATCAGGGGGAGGATTCTAACGCAAGCCTTAAA AAATGCTAGTAAGCTCTGCTTGACAGCTGATGTCCTAGGAGAATGCACCCAATCATCATGTTCTAATCTTGCAACT ACTGTCATGAGGTTAACTGAGAATGGTGTTGAAAAGATATCTGTTTCTACTTGAATATCTATATGACCATCAAAC AGCTCTCCTATGATATCATCTTCCCTCCAAGTGTCAATTCCTGGAGATCAGATCACATGAGAATACATAAATAATCC ACACCTGGTATCACGATTGGCTCTTTTGCCATCCCAGTTAGGAGGTCTAAACTACCTGTCATGCAGTAGGCTGTTC AATCGAAACATAGGCGACCCGGTGGTTTCCGCAGTTGCAGATCTTAAGAGATTAATTAAATCAGGATGTATGGATT ACTGGATCCTTTATAACTTATTAGGGAGAAACCGGGAAACGGCTCATGGGCTACTTTAGCAGCTGACCCGTACTC AATCAATATAGAGTATCAATACCCTCCAACTACAGCTCTTAAGAGGGCACACCCAACAAGCTCTGATGGAACTCAGT ACGAATCCAATGTTACGTGGCATATTCTCTGACAATGCACAGGCAGAAGAAATAACCTTGCTAGGTTTCTCCTGG ATAGGGAGGTGATCTTTCCGCGTGTAGCTCACATCATCATTGAGCAAACCAGTGTCGGGAGGAGAAAACAGATTCA AGGATATTTGGATTCAACTAGATCGATAATGAGGAAATCACTAGAAATTAAGCCCTTATCCAATAGGAAGCTTAAT GAAATACTGGATTACAACATCAATTACCTAGCTTACAATTTGGCATTACTCAAGAATGCTATTGAACCTCCGACTT ATTTGAAGGCAATGACACTTGAAACATGTAGCATCGACATTGCAAGGAACCTCCGGAAGCTCTCCTGGGCCCCACT CTTGGGTGGGAGAAATCTTGAAGGATTAGAGACGCCAGATCCCATTGAAATTACTGCAGGAGCATTAATTGTTGGA TCGGGCTACTGTGAACAGTGTGCTGCAGGAGACAATCGATTCACATGGTTTTTCTTGCCATCTGGTATCGAGATAG GAGGGGATCCCCGTGATAATCCTCCTATCCGTGTACCGTACATTGGCTCCAGGACTGATGAGAGGAGGGTAGCCTC AATGGCATACATCAGGGGTGCCTCGAGTAGCTTAAAAGCAGTTCTTAGACTGGCGGGAGTGTACATCTGGGCATTC GGAGATACTCTGGAGAATTGGATAGATGCACTGGATTTGTCTCACACTAGAGTTAACATCACACTTGAACAGCTGC AATCCCTCACCCCACTTCCAACCTCTGCCAATCTAACCCATCGGTTGGATGATGGCACAACTACCCTAAAGTTTAC TCCTGCGAGCTCTTACACCTTTTCAAGTTTCACTCATATATCAAATGATGAGCAATACCTGACAATTAATGACAAA ACTGCAGATTCAAATATAATCTACCAACAGTTAATGATCACTGGACTCGGAATCTTAGAAACATGGAATAATCCCC CAATCAATAGAACATTCGAAGAATCTACCCTACATTTGCACACTGGTGCATCATGTTGTGTCCGACCTGTGGACTC CTGCATTCTCTCAGAAGCATTAACAGTCAAGCCACATATTACAGTACCGTACAGCAATAAATTTGTATTTGATGAG GACCCGCTATCTGAATATGAAACTGCAAAACTGGAATCGTTATCATTCCAAGCCCAATTAGGCAACATTGATGCTG TAGATATGACAGGTAAATTAACATTATTGTCCCAATTCACTGCAAGGCAGATTATCAATGCAATCACTGGACTCGA TGAGTCTGTCTCTCTTACTAATGATGCCATTGTTGCATCAGACTATGTCTCCAATTGGATTAGTGAATGCATGTAT ACCAAATTAGATGAATTATTTATGTATTGTGGGTGGGAACTACTATTGGAACTATCCTATCAAATGTATTATCTGA GGGTAGTTGGGTGGAGTAATATAGTGGATTATTCTTACATGATCTTGAGAAGAATCCCGGGTGCAGCATTAAACAA TCTGGCATCTACATTAAGTCATCCAAAACTTTTCCGACGAGCTATCAACCTAGATATAGTTGCCCCCTTAAATGCT CCTCATTTTGCATCTCTGGACTACATCAAGATGAGTGTGGATGCAATACTCTGGGGCTGTAAAAGAGTCATCAATG TGCTCTCCAATGGAGGGGACTTAGAATTAGTTGTGACATCTGAAGATAGCCTTATTCTCAGTGACCGATCCATGAA TCTCATTGCAAGGAAATTAACTTTATTATCACTGATTCACCATAATGGTTTGGAACTACCAAAGATTAAGGGGTTC TCTCCTGATGAGAAGTGTTTCGCTTTGACAGAATTTTTGAGGAAAGTGGTGAACTCAGGGTTGAGTTCAATAGAGA ACCTATCAAATTTTATGTACAATGTGGAGAACCCACGGCTTGCAGCATTCGCCAGCAACAATTACTACCTGACCAG AAAATTATTGAATTCAATACGAGATACTGAGTCGGGTCAAGTAGCAGTCACCTCATATTATGAATCATTAGAATAT ATTGATAGTCTTAAGCTAACCCCACATGTGCCTGGCACCTCATGCATTGAGGATGATAGTCTATGTACAAATGATT ACATAATCTGGATCATAGAGTCTAATGCAAACTTGGAGAAGTATCCAATTCCAAATAGCCCTGAGGATGATTCCAA TTTCCATAACTTTAAGTTGAATGCTCCATCGCACCATACCTTACGCCCATTAGGGTTGTCATCAACTGCTTGGTAT AAGGGTATAAGCTGCTGCAGGTACCTTGAGCGATTAAAGCTACCACAAGGTGATCATTTATATATTGCAGAAGGTA GTGGTGCCAGTATGACAATCATAGAATACCTATTCCCAGGAAAAAGATATATTACATTCTTTATTTAGTAGTGG TGACAATCCCCCCACAAAGAAATTATGCACCAATGCCTACTCAGTTCATTGAGAGTGTCCCATACAAGCTCTGGCAA GCACACACAGATCAATCCCGAGATTTTTGAGGACTTCCATCCCTCTATGGAACGGAAACGCCGCCATGACTGACA TAGGAATGACAGCTTGTGTGAGAATTCATCATCAATCGAGTCGGCCCAAGGACTTGCAGTTTAGTACATGTAGATTT GGAATCAAGTGCAAGCTTAAATCAACAATGCCTGTCAAAGCCGATAATTAATGCTATCATCACTGCTACAACTGTT TTGTGCCCTCATGGGGTGCTTATTCTGAAATATAGTTGGTTGCCATTTACTAGATTTAGTACTTTGATCACTTTCT TATGGTGCTACTTTGAGAGAATCACTGTTCTTAGGAGCACATATTCTGATCCAGCTAATCATGAGGTTTTATTTAAT TTGTATCCTTGCCAACAACTTTGCATTCCAGACTGTCTCGCAGGCAACAGGAATGGCGATGACTTTAACTGATCAA GGGTTTACTTTGATATCACCTGAAAGAATAAATCAGTATTGGGATGGTCACTTGAAGCAAGAACGTATCGTAGCAG AAGCAATTGATAAGGTGGTTCTAGGAGAAAATGCTCTATTTAATTCGAGTGATAATGAATTAATTCTCAAATGTGG AGGGACACCAAATGCACGGAATCTCATCGATATCGAGCCAGTCGCAACTTTCATAGAATTTGAACAATTGATCTGC ACAATGTTGACAACCCACTTGAAGGAAATAATTGATATAACAAGGTCTGGAACCCAGGATTATGAAAGTTTATTAC TCACTCCTTACAATTTAGGTCTTCTTGGTAAAATCAGTACGATAGTGAGATTATTAACAGAAAGGATTCTAAATCA TACTATCAGGAATTGGTTGATCCTCCCACCTTCGCTCCGGATGATCGTGAAGCAGGACTTGGAATTCGGCATATTC AGGATTACTTCCATCCTCAATTCTGATCGGTTCCTGAAGCTTTCTCCAAATAGGAAATACTTGATTGCACAATTAA CTGCAGGCTACATTAGGAAATTGATTGAGGGGGATTGCAATATCGATCTAACCAGACCTATCCAAAAGCAAATCTG GAAAGCATTAGGTTGTGTAGTCTATTGTCACGATCCAATGGATCAAAGGGAGTCAACAGAGTTTATTGATATAAAT ATTAATGAAGAAATAGACCGCGGGATCGATGGCGAGGAAATCTAAACATATCAAGAATCAGAATTAGTTTAAGAAA AAAGAAGAGGATTAATCTTGGTTTTCCCCTTGGT

[0116] The underlined sequence is the PIV5 genomic nucleic acid sequence. The italicized and underlined sequence is the entire Pal nucleic acid sequence. The double-underlined sequence is the AOS-modified NTTM-Pal nucleic acid sequence.

[0117] c. pMHD35- PIV5-NTTM-OspA BPBPk

[0118] This article provides pMHD35-PIV5-NTTM-OspA BPBPk The nucleic acid sequences from 5' to 3' are as follows:

[0119] ACCAAGGGGAAAATGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCTA TGGCCTTCGTGACCGACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCGACA CAAAAATGTCATCCGTGCTTAAAGCATATGAGCGATTCACGCTCACTCAAGAACTGCAAGATCAGAGTGAGGAAGGT ACAATCCCACCTACAACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGATCCCG GCTTCTTCTATTCTGCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACTCACAA TGTTTTCGCTACCATCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATCGAAAGG GTAGAGATCGATGGCTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCTCGTTCAGGTATGAGCCGTGGAGAGAT CAATGCCTATGCTGCACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTCGTTGATTCCGAAGTCG AGGGAACTGCATGGGATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGGATAGTGACT TGCAAGTGCATGACTGCGCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAATATCGTCAGCAAGGCAG GATCAACCCGAGATATCTCCTGCAACCGGAGGCTCGACGAATAATCCAGAATGTAATCCGGAAGGGAATGGTGGTCA GACATTTCCTCACCTTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTATGCTATGGTAGGG GATGTTGGAAAGTATATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAATATGCATTAGGAACTAGATG GCCCACACTTGCTTTAGCTGCATTTTCAGGAGAGCTAACAAAGCTAAAGTCCCTCATGGCATTATACCAGACCCTTG GTGAGCAGGCCCGATATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTTGCTGCAGCAAACTACCCACTGCTA TATAGCTATGCTATGGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTCTCCAGATCATACATGAA CAAGACATATTTCCAATTGGGAATGGAAACTGCAAGAAAACAACAGGGTGCAGTTGACATGAGGATGGCAGAAGATC TCGGTCTAACTCAAGCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACAGCAAATCGAGGGGCAGAC ACCAGGGGAGGAGTCAACCCGTTCTCATCTGTCACTGGGACAACTCAGGTGCCCGCTGCAGCAACAGGTGACACACT CGAGAGTTACATGGCAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCATGATGATGAGATGCCACCAT TGGAAGAGGAGGAAGAGGACGACACATCTGCAGGTCCACGCACTGGACCAACTCTTGAACAAGTGGCCTTGGACATC CAGAACGCAGCAGTTGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGGGTGATCTTGACATCTAGACAAT TCAGATCCCAATCTAAAATTGACATACCTAATTGATTAGTTAGATGGAACTACAGTGGATTCCATAAGGTTCCTGCC TACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGGTGCCAACAGCGCAATCCA CAATCTACAATGGATCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATAAGCTCATAGAGACAGGCCTGAATAC TGTAGAGTATTTTACTTCCCAACAAGTCACAGGAACATCCTCTCTTGGAAAGAATACAATACCACCAGGGGTCACAG GACTACTAACCAATGCTGCAGAGGCAAAGATCCAAGAGTCAACTAACCATCAGAAGGGCTCAGTTGGTGGGGGTGCA AAACCAAAGAAACCGCGACCAAAAATTGCCATTGTGCCAGCAGATGACAAAACAGTGCCCGGAAAGCCGATCCCAAA CCCTCTATTAGGTCTGGACTCCACCCCGAGCACCCAAACTGTGCTTGATCTAAGTGGGAAAACATTACCATCAGGAT CCTATAAGGGGGTTAAGCTTGCGAAATTTGGAAAAGAAAATCTGATGACACGGTTCATCGAGGAACCCAGAGAGAAT CCTATCGCAACCAGTTCCCCCATCGATTTTAAGAGGGGCAGGGATACCGGCGGGTTCCATAGAAGGGAGTACTCAAT CGGATGGGTGGGAGATGAAGTCAAGGTCACTGAGTGGTGCAATCCATCCTGTTCTCCAATCACCGCTGCAGCAAGGC GATTTGAATGCACTTGTCACCAGTGTCCAGTCACTTGCTGAATGTGAACGAGATACTTAATACAGTGAGAAATTT GGACTCTCGGATGAATCAACTGGAGACAAAAGTAGATCGCATTCTCTCATCTCAGTCTAATCCAGACCATCAAGA ATGACATAGTTGGACTTAAAGCAGGGATGGCTACTTTAGAAGGAATGATTACAACTGTGAAAATCATGGACCCGGGA GTTCCCAGTAATGTTACTGTGGAAGATGTACGCAAGACACTAAGTAACCATGCTGTTGTTGTGCCAGAATCATTCAA TGATAGTTTCTTGACTCAATCTGAAGATGTAATTTCACTTGATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAGA AGATTGTCAGGAAGGTTCCTCCTCAGAAGGATCTGACTGGATTGAAGATTACACTAGAGCAATTGGCAAAGGATTGC ATCAGCAAACCGAAGATGAGGGAAGAGTATCTCCTCAAAATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCT CAAGAAAGCAATCATCCGCAGTGCAATTTGATCAAGAAACACCCAATTACACTACACTGGTATGACACTGTACTAAC CCTGAGGGTTTTAGAAAAACACGATTAACGATAAATAAGCCCGAACACTACACACTACCTGAGGCAGCCATGCCATCC ATCAGCATTCCCGCAGACCCCACCAATCCACGTCAATCAATAAAAGCGTTCCCAATTGTGATCAACAGTGATGGGGG TGAGAAAGGCCGCTTGGTTAAACAACTACGCACAACCTACTTGAATGACCTAGATACTCATGAGCCACTGGTGACAT TCATAAATACCTATGGATTCATCTACGAACAGGATCGGGGGAATACCATTGTCGGAGAGGATCAACTTGGGAAGAAA AGAGAGGCTGTGACCGCTGCAATGGTTACCCTTGGATGTGGGCCTAATCTACCATCATTAGGGAATGTCCTGGGACA ACTGAGGGAATTCCAGGTCACTGTTAGGAAGACATCCAGCAAAGCGGAAGAGATGGTCTTTGAAATTGTTAAGTATC CGAGAATATTTCGGGGTCATACATTAATCCAGAAAGGACTAGTCTGTGTCTCCGCAGAAAAATTTGTTAAGTCACCA GGGAAAATACAATCTGGAATGGACTATCTCTTCATTCCGACATTTCTGTCAGTGACTTACTGTCCAGCTGCAATCAA ATTTCAGGTACCTGGCCCCATGTTGAAAATGAGATCAAGATACACTCAGAGCTTACAACTTGAACTAATGATAAGAA TCCTGTGTAAGCCCGATTCGCCACTTATGAAGGTCCATACCCCTGACAAGGAGGGAAGAGGATGTCTTGTATCAGTA TGGCTGCATGTATGCAACATCTTCAAATCAGGAAACAAGAATGGCAGTGAGTGGCAGGAATACTGGATGAGAAAGTG TGCTAACATGCAACTTGAAGTGTCGATTGCAGATATGTGGGGACCAACTATCATAATTCATGCCAGAGGTCACATTC CCAAAAGTGCTAAGTTGTTTTTTGGAAAGGGTGGATGGAGCTGCCATCCACTTCACGAAGTTGTTCCAAGTGTCACT AAAACACTATGGTCCGTGGGCTGTGAGATTACAAAGGCGAAGGCAATAATACAAGAGAGTAGCATCTCTCTTCTCGT GGAGACTACTGACATCATAAGTCCAAAAGTCAAAATTTCATCTAAGCATCGCCGCTTTGGGAAATCAAATTGGGGTC TGTTCAAGAAAACTAAATCACTGCCTAACCTGACGGAGCTGGAATGACTGACCTCTAATCGAGACTACACCGCCGCA AACTATAGGTGGGTGGTACCTCAGTGATTAATCTTGTAAGCACTGATCGTAGGCTACAACACACTAATATTATCCAG ATTAGAGAGCTTAATTAGCTCTGTATTAATAATAACACTACTATTCCAATAACTGGAATCACCAGCTTGATTTATCT CCAAAATGATTCAAAGAAAACAAATCATATTAAGACTATCCTAAGCACGAACCCATATCGTCCTTCAAATCATGGGT ACTATAATTCAATTTCTGGTGGTCTCCTGTCTATTGGCAGGAGCAGGCAGCCTTGATCCAGCAGCCCTCATGCAAAT CGGTGTCATTCCAACAAATGTCCGGCAACTTATGTATTATACTGAAGCTTCATCAGCATTCATTGTTGTGAAGTTAA TGCCTACAATTGACTCGCCGATTAGTGGATGTAATATAACATCAATTTCAAGCTATAATGCAACAGTGACAAAACTC CTACAGCCGATCGGTGAGAATTTGGAGACGATTAGGAACCAGTTGATTCCAACTCGGAGGAGACGCCGGTTTGCAGG GGTGGTGATTGGATTAGCTGCATTAGGAGTAGCTACTGCCGCACAGGTCACTGCCGCAGTGGCACTAGTAAAGGCAA ATGAAAATGCTGCGGCTATACTCAATCTCAAAAATGCAATCCAAAAAACAAATGCAGCAGTTGCAGATGTGGTCCAG GCCACACAATCACTAGGAACGGCAGTTCAAGCAGTTCAAGATCACATAAACAGTGTGGTAAGTCCAGCAATTACAGC AGCCAATTGTAAGGCCCAAGATGCTATCATTGGCTCAATCCTCAATCTCTATTTGACCGAGTTGACAACCATCTTCC ACAATCAAATTACAAACCCTGCATTGAGTCCCATTACAATTCAAGCTTTAAGGATCCTACTGGGGAGTACCTTGCCG ACTGTGGTCGAAAAATCTTTCAATACCCAGATAAGTGCAGCTGAGCTTCTCTCATCAGGGTTATTGACAGGCCAGAT TGTGGGATTAGATTTGACCTATATGCAGATGGTCATAAAAATTGAGCTGCCAACTTTAACTGTACAACCTGCAACCC AGATCATAGATCTGGCCACCATTTCTGCATTCATTAACAATCAAGAAGTCATGGCCCAATTACCAACACGTGTTATG GTGACTGGCAGCTTGATCCAAGCCTATCCCGCATCGCAATGCACCATTACACCCAACACTGTGTACTGTAGGTATAA TGATGCCCAAGTACTCTCAGATGATACTATGGCTTGCCTCCAAGGTAACTTGACAAGATGCACCTTCTCTCCAGTGG TTGGGAGCTTTCTCACTCGATTCGTGCTGTTCGATGGAATAGTTTATGCAAATTGCAGGTCGATGTTGTGCAAGTGC ATGCAACCTGCTGCTGTGATCCTACAGCCGAGTTCATCCCCTGTAACTGTCATTGACATGTACAAATGTGTGAGTCT GCAGCTTGACAATCTCAGATTCACCATCACTCAATTGGCCAATGTAACCTACAATAGCACCATCAAGCTTGAATCAT CCCAGATCTTGTCTATTGATCCGTTGGATATATCCCAGAATCTAGCTGCGGTGAATAAGAGTCTAAGTGATGCACTA CAACACTTAGCACAAAGTGACACATATCTTTCTGCAATCACATCAGCTACGACTACAAGTGTATTATCCATAATAGC AATCTGTCTTGGATCGTTAGGTTTAATATTAATAATCTTGCTCAGTGTAGTTGTGTGGAAGTTATTGACCATTGTCG TTGCTAATCGAAATAGAATGGAGAATTTTGTTTATCATAAATAAGCATTCCACCACTCACGATCTGATCTCAGTGAG AAAAATCAACCTGCAACTCTTGGAACAAGATAAGACAGTCATCCATTAGTAATTTTTAAGAAAAAAACGATAGGACC GAACCTAGTATTGAAAGAACCGTCTCGGTCAATCTAGGTAATCGAGCTGATACCGTCTCGGAAAGCTCAAATCATGC TGCCTGATCCGGAAGATCCGGAAAGCAAGAAAGCTACAAGGAGAGCAGGAAACCTAATTATCTGCTTCCTATTCATC TTCTTTCTGTTTGTAACCTTCATTGTTCCAACTCTAAGACACTTGCTGTCCTAACACCTGCTATAGGCTATCCACTG CATCATCTTACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGAC

[0120]

[0121] GCCTGATCCTGGCCCTGATCGCCTGCAAGCAGAACGTGAGCTCCCTGGACGAGAAGAATAGCGTGTCC GTGGATCTGCCCGGCGAGATGAAGGTGCTGGTGAGCAAGGAGAAGAACAAGGACGGCAAGTATGATCTGATCGCCA CAGTGGACAAGCTGGAGCTGAAGGGCACCAGCGATAAGAACAATGGATCCGGCGTGCTGGAGGGAGTGAAGGCAGA CAAGTCTAAGGTGAAGCTGACCATCAGCGACGATCTGGGCCAGACCACACTGGAGGTGTTCAAGGAGGACGGCAAG ACCCTGGTGTCCAAGAAGGTGACATCTAAGGATAAGTCTAGCACCGAGGAGAAGTTTAACGAGAAGGGCGAGGTGT CCGAGAAGATCATCACACGGGCGACGCACCAGACTGGAGTACACAGGCATCAAGTCCGATGGCTGCTGGCAAGGC CAAGGAGGTGCTGAAGGGCTTCACACTGGAGGGCAAGGTGGCCAATGATAAGGTGACCCTGGAGGTGAAGGAGGGC ACCGTGACACTGTCCAAGAACATCTCTAAGAGCGGCGAGGTGTCTGTGGAGCTGAATGACACCGATTCCTCTGCCG CCAAAAAAGACCGCCGCCTGGAACAGCAAGACCTCCACACTGACCATCTCTGTGAATAGCAAGAAGACCACACA GCTGGTGTTTACAAAGCAGGACACAATCACCGTGCAGAAGTATGATAGCGCCGGAACCAACCTGGAGGGAACAGCA GTGGAGATCAAGACCCTGGACGAGCTGAAGGAATGCCCTGAAGTGAACGCGTGCTAGCTCTCCTGCCACTACTTCCTA CTCACATCATATCTATTTAAAAGAAAAAATAGGCCCGAACACTAATCGTGCCGGCAGTGCCACTGCACACACAACA CTACACATACAATACACTACAATGGTTGCAGAAGATGCCCCTGTTAGGGCCACTTGCCGAGTATTATTTCGAACAA CAACTTTAATCTTTCTATGCACACTACTAGCATTAAGCATCTCTATCCTTTATGAGAGTTTAATAACCCAAAAGCA AATCATGAGCCAAGCAGGCTCAACTGGATCTAATTCTGGATTAGGAAGTATCACTGATCTTCTTAATAATATTCTC TCTGTCGCAAATCAGATTATATATAACTCTGCAGTCGCTCTACCTCTACAATTGGACACTCTTGAATCCAACACTCC TTACAGCCATTAAGTCTCTTCAAACCAGTGACAAGCTAGAACAGAACTGCTCGTGGAGTGCTGCACTGATTAATGA TAATAGATACATTAATGGCATCAATCAGTTCTATTTTTCAATTGCTGAGGGTCGCAATCTGACACTTGGCCCACTT CTTAATATGCCTAGTTTCATTCCAACTGCCACGACACCAGAGGGCTGCACCAGGATCCCATCATTCTCGCTCACTA AGACACACTGGTGTTATACACACAATGTTATCCTGAATGGATGCCAGGATCATGTATCCTCAAATCAATTTGTTTC TATGGGAATCATTGAACCCACTTCTGCCGGGTTTCCATTCTTTCGAACCCTAAAGACTCTATATCTCAGCGATGGG GTCAATCGTAAGAGCTGCTCTATCAGTACAGTTCCGGGGGGTTGTATGATGTACTGTTTTGTTTCTACTCAACCAG AGAGGGATGACTACTTTTCTGCCGCTCCTCCAGAACAACGAATTATTATAATGTACTATAATGATACAATCGTGGA GCGCATAATTAATCCACCCGGGGTACTAGATGTATGGGCAACATTGAACCCAGGAACAGGAAGCGGGGTATATTAT TTAGGTTGGGTGCTCTTTCCAATATATGGCGGCGTGATTAAAGGTACGAGTTTATGGAATAATCAAGCAAATAAAT ACTTTATCCCCCAGATGGTTGCTGCTCTCTGCTCACAAAACCAGGCAACTCAAGTCCAAAATGCTAAGTCATCATA CTATAGCAGCTGGTTTGGCAATCGAATGATTCAGTCTGGGATCCTGGCATGTCCTCTTCGACAGGATCTAACCAAT GAGTGTTTAGTTCTGCCCTTTTCTAATGATCAGGTGCTTATGGGTGCTGAAGGGAGATTATACATGTATGGTGACT CGGTGTATTACTATCAAAGAAGCAATAGTTGGTGGCCTATGACCATGCTGTATAAGGTAACCATAACATTCACTAA TGGTCAGCCATCTGCTATATCAGCTCAGAATGTGCCCACACAGCAGGTCCCTAGACCTGGGACAGGAGACTGCTCT GCAACCAATAGATGTCCCGGTTTTTGCTTGACAGGAGTGTATGCCGATGCCTGGTTACTGACCAACCCTTCGTCTA CCAGTACATTTGGATCAGAAGCAACCTTCACTGGTTCTTATCTCAACACAGCAACTCAGCGTATCAATCCGACGAT GTATATCGCGAACAACACACAGATCATAAGCTCACAGCAATTTGGATCAAGCGGTCAAGAAGCAGCATATGGCCAC ACAACTTGTTTTAGGGACACAGGCTCTGTTATGGTATACTGTATCTATATTATTGAATTGTCCTCATCTCTCTTAG GACAATTTCAGATTGTCCCATTTATCCGTCAGGTGACACTATCCTAAAGGCAGAAGCCTTCAGGTCTGACCCAGCC AATCAAAGCATTATACCAGACCATGGAATGCATACCAAACATTATTGACACTAATGACACACAAAATTGGTTTTAA GAAAAACCAAGAGAACAATAGGCCAGAATGGCTGGGTCTCGGGAGATATTACTCCCTGAAGTCCATCTCAATTCAC CAATTGTAAAGCATAAGCTATACTATTACATTCTACTTGGAAACCTCCCAAATGAGATCGACCTTGACGATTTAGG TCCATTACATAATCAAAATTGGAATCAGATAGCACATGAAGAGTCTAACTTAGCTCAACGCTTGGTAAATGTAAGA AATTTTCTAATTACCCACATCCCTGATCTTAGAAAGGGCCATTGGCAAGAGTATGTCAATGTAATACTGTGGCCGC GAATTCTTCCCTTGATCCCGGATTTTAAAATCAATGACCAATTGCCTCTGCTCAAAAATTGGGACAAGTTAGTTAA AGAATCATGTTCAGTAATCAATGCAGGTACTTCCCAGTGCATTCAGAATCTCAGCTATGGACTGACAGGTCGTGGG AACCTCTTTACACGATCACGTGAACTCTCTGGTGACCGCAGGGATATTGATCTTAAGACAGTTGTGGCAGCATGGC ATGACTCAGACTGGAAAAGAATAAGTGATTTTTGGATTATGATCAAATTCCAGATGAGACAATTAATTGTTAGGCA AACAGATCATAATGATTCTGATTTAATCACGTATATCGAAAATAGAGAAGGCATAATCATCATAACCCCTGAACTG GTAGCATTATTTAACACTGAGAATCATACACTAACATACATGACCTTTGAAATTGTACTGATGGTTTCAGATATGT ACGAAGGTCGTCACAACATTTTATCACTATGCACAGTTAGCACTTACCTGAATCCTCTGAAGAAAAGAATAACATA TTTATTGAGCCTTGTAGATAACTTAGCTTTTCAGATAGGTGATGCTGTATATAACATAATTGCTTTGCTAGAATCC TTTGTATATGCACAGTTGCAAATGTCAGATCCCATCCCAGAACTCAGAGGACAATTCCATGCATTCGTATGTTCTG AGATTCTTGATGCACTAAGAGGAACTAATAGTTTCACCCAGGATGAATTAAGAACTGTGACAACTAATTTGATATC CCCATTCCAAGATCTGACCCCAGATCTTACGGCTGAATTGCTCTGTATAATGAGGCTTTGGGGACACCCCATGCTC ACTGCCAGTCAAGCTGCAGGAAAGGTACGCGAGTCTATGTGTGCTGGAAAAGTATTAGACTTTCCCACCATTATGA AAACACTAGCCTTTTTCCATACTATTCTGATCAATGGATACAGGAGGAAGCATCATGGAGTATGGCCACCCTTAAA CTTACCGGGTAATGCTTCAAAGGGTCTCACGGAACTTATGAATGACAATACTGAAATAAGCTATGAATTCACACTT AAGCATTGGAAGGAAGTCTCTCTTATAAAATTCAAGAAATGTTTTGATGCAGACGCAGGTGAGGAACTCAGTATAT TTATGAAAGATAAGGCAATTAGTGCCCCAAAACAAGACTGGATGAGTGTGTTTAGAAGAAGCCTAATCAAACAGCG CCATCAGCATCATCAGGTCCCCCTACCAAATCCATTCAATCGACGGCTGTTGCTAAACTTTCTCGGAGATGACAAA TTCGACCCGAATGTGGAGCTACAGTATGTAACATCAGGTGAGTATCTACATGATGACACGTTTTGTGCATCATATT CACTAAAAGAGAAGGAAATTAAACCTGATGGTCGAATTTTTGCAAAGTTGACTAAGAGAATGAGATCATGTCAAGT TATAGCAGAATCTCTTTTAGCGAACCATGCTGGGAAGTTAATGAAAGAGAATGGTGTTGTGATGAATCAGCTATCA TTAACAAAATCACTATTAACAATGAGTCAGATTGGAATAATATCCGAGAAAGCTAGAAAGTCAACTCGAGATAACA TAAATCAACCTGGTTTCCAGAATATCCAGAGAAATAAATCACATCACTCCAAGCAAGTCAATCAGCGAGATCCAAG TGATGACTTTGAATTGGCAGCATCTTTTTTAACTACTGATCTCAAAATATTGTTTACAATGGAGGTACCAGACA ATTATCCCATTTGCTCAATCATTAAACAGAATGTATGGTTATTCCTCATCTCTTTGAGTGGATTCACTTACGGCTAA TGCGTAGTACACTTTACGTGGGGGATCCCTTCAACCCACCAGCAGATACCAGTCAATTTGATCTAGATAAAGTAAT TAATGGAGATATCTTCATTGTATCACCCAGAGGTGGAATTGAAGGGCTGTGTCAAAAGGCTTGGACAATGATATCT ATCGCTGTGATAATTCTATCTGCCACAGAGTCTGGCACACGAGTAATGAGTATGGTGCAGGGAGATAATCAAGCAA TTGCTGTCACCACACGAGTACCAAGGAGCCTGCCGACTCTTGAGAAAAAGACTATTGCTTTTTAGATCTTGTAATCT ATTCTTTGAGAGGTTAAAATGTAATAATTTTGGATTAGGTCACCATTTGAAAGAACAAGAGACTATCATTAGTTCT CACTTCTTTGTTTATAGCAAGAGAATATTCTATCAGGGGAGGATTCTAACGCAAGCCTTAAAAAATGCTAGTAAGC TCTGCTTGACAGCTGATGTCCTAGGAGAATGCACCCAATCATCATGTTCTAATCTTGCAACTACTGTCATGAGGTT AACTGAGAATGGTGTTGAAAAAGATATCTGTTTCTACTTGAATATCTATATGACCATCAAACAGCTCTCCTATGAT ATCATCTTCCCTCAAGTGTCAATTCCTGGAGATCAGATCACATTAGAATACATAAATAATCCACACCTGGTATCAC GATTGGCTCTTTTGCCATCCCAGTTAGGAGGTCTAAACTACCTGTCATGCAGTAGGCTGTTCAATCGAAACATAGG CGACCCGGTGGTTTCCGCAGTTGCAGATCTTAAGAGATTAATTAAATCAGGATGTATGGATTACTGGATCCTTTAT AACTTATTAGGGAGAAAACCGGGAAACGGCTCATGGGCTACTTTAGCAGCTGACCCGTACTCAATCAATATAGAGT ATCAATACCCTCCAACTACAGCTCTTAAGAGGCACACCCAACAAGCTCTGATGGAACTCAGTACGAATCCAATGTT ACGTGGCATATTCTCTGACAATGCACAGGCAGAAGAAAATAACCTTGCTAGGTTTCTCCTGGATAGGGAGGTGATC TTTCCGCGTGTAGCTCACATCATCATTGAGCAAACCAGTGTCGGGAGGAGAAAACAGATTCAAGGATATTTGGATT CAACTAGATCGATAATGAGGAAATCACTAGAAATTAAGCCCTTATCCAATAGGAAGCTTAATGAAATACTGGATTA CAACATCAATTACCTAGCTTACAATTTGGCATTACTCAAGAATGCTATTGAACCTCCGACTTATTTGAAGGCAATG ACACTTGAAACATGTAGCATCGACATTGCAAGGAACCTCCGGAAGCTCTCCTGGGCCCCACTCTTGGGTGGGAGAA ATCTTGAAGGATTAGAGACGCCAGATCCCATTGAAATTACTGCAGGAGCATTAATTGTTGGATCGGGCTACTGTGA ACAGTGTGCTGCAGGAGACAATCGATTCACATGGTTTTTCTTGCCATCTGGTATCGAGATAGGAGGGGATCCCCGT GATAATCCTCCTATCCGTGTACCGTACATTGGCTCCAGGACTGATGAGAGGAGGGTAGCCTCAATGGCATACATCA GGGGTGCCTCGAGTAGCTTAAAAGCAGTTCTTAGACTGGCGGGAGTGTACATCTGGGCATTCGGAGATACTCTGGA GAATTGGATAGATGCACTGGATTTGTCTCACACTAGAGTTAACATCACACTTGAACAGCTGCAATCCCTCACCCCA CTTCCAACCTCTGCCAATCTAACCCATCGGTTGGATGATGGCACAACTACCCTAAAGTTTACTCCTGCGAGCTCTT ACACCTTTTCAAGTTTCACTCATATATCAAATGATGAGCAATACCTGACAATTAATGACAAAACTGCAGATTCAAA TATAATCTACCAACAGTTAATGATCACTGGACTCGGAATCTTAGAAACATGGAATAATCCCCCAATCAATAGAACA TTCGAAGAATCTACCCTACATTTGCACACTGGTGCATCATGTTGTGTCCGACCTGTGGACTCCTGCATTCTCTCAG AAGCATTAACAGTCAAGCCACATATTACAGTACCGTACAGCAATAAATTTGTATTTGATGAGGACCCGCTATCTGA ATATGAAACTGCAAAACTGGAATCGTTATCATTCCAAGCCCAATTAGGCAACATTGATGCTGTAGATATGACAGGT AAATTAACATTATTGTCCCAATTCACTGCAAGGCAGATTATCAATGCAATCACTGGACTCGATGAGTCTGTCTCTC TTACTAATGATGCCATTGTTGCATCAGACTATGTCTCCAATTGGATTAGTGAATGCATGTATACCAAATTAGATGA ATTATTTATGTATTGTGGGTGGGAACTACTATTGGAACTATCCTATCAAATGTATTATCTGAGGGTAGTTGGGTGG AGTAATAGTGGATTATTCTTACATGATCTTGAAGAATCCCGGGTGCAGCATTAAACAATCTGGCATCTACAT TAAGTCATCCAAAACTTTTCCGACGAGCTATCAACCTAGATATAGTTGCCCCCTTAAATGCTCCTCATTTTGCATC TCTGGACTACATCAAGATGAGTGTGGATGCAATACTCTGGGGCTGTAAAAGAGTCATCAATGTGCTCTCCAATGGA GGGGACTTAGAATTAGTTGTGACATCTGAAGATAGCCTTATTCTCAGTGACCGATCCATGAATCTCATTGCAAGGA AATTAACTTTATTCACTGATTCACCATAATGGTTTGGAACTACCAAAGATTAAGGGTTCTCTCCTGATGAGAA GTGTTTCGCTTTGACAGAATTTGAGGAAAGTGGTGAACTCAGGGTTGAGTTCAATAGAGAACCTATCAAATTTT ATGTACAATGTGGAGAACCCACGGCTTGCAGCATTCGCCAGCAACAATTACTACCTGACCAGAAAATTATTGAATT CAATACGAGATACTGAGTCGGGTCAAGTAGCAGTCCACCTCATATTATGAATCATTAGAATATTGATAGTCTTAA GCTAACCCCACATGTGCCTGGCACCTCATGCATTGAGGATGATAGTCTATGTACAAATGATTACATAATCTGGATC ATAGAGTCTAATGCAAACTTGGAGAAGTATCCAATTCCAAATAGCCCTGAGGATGATTCCAATTTCCATAACTTTA AGTTGAATGCTCCATCGCACCATACCTTACGCCCATTAGGGTTGTCATCAACTGCTTGGTATAAGGGTATAAGCTG CTGCAGGTACCTTGAGCGATTAAAGCTACCACAAGGTGATCATTTATATTGCAGAAGGTAGTGGTGCCAGTATG ACAATCATAGAATACCTATTCCCAGGAAAAGATATTACAATTCTTTATTTAGTAGTGGTGACAATCCCCCAC AAAGAAATTATGCACCAATGCCTACTCAGTTCATTGAGAGTGTCCCATACAAGCTCTGGCAAGCACACACAGATCA ATATCCCGAGATTTTTGAGGACTTCATCCCTCTATGGAACGGAAACGCCGCCATGACTGACATAGGAATGACAGCT TGTGTAGAATTCATCATCAATCGAGTCGGCCCAAGGACTTGCAGTTTAGTACATGTAGATTTGGAATCAAGTGCAA GCTTAAATCAACAATGCCTGTCAAAGCCGATAATTAATGCTATCATCACTGCTACAACTGTTTTGTGCCCTCATGG GGTGCTTATTCTGAAATATAGTTGGTTGCCATTTACTAGATTTAGTACTTTGATCACTTTCTTATGGTGCTACTTT GAGAGAATCACTGTTCTTAGGAGCACATATTCTGATCCAGCTAATCATGAGGTTTATTTAATTTGTATCCTTGCCA ACAACTTTGCATTCCAGACTGTCTCGCAGGCAACAGGAATGGCGATGACTTTAACTGATCAAGGGTTTACTTTGAT ATCACCTGAAAGAATAAATCAGTATTGGGATGGTCACTTGAAGCAAGAACGTATCGTAGCAGAAGCAATTGATAAG GTGGTTCTAGGAGAAAATGCTCTATTTAATTCGAGTGATAATGAATTAATTCTCAAATGTGGAGGGACACCAAATG CACGGAATCTCATCGATATCGAGCCAGTCGCAACTTTCATAGAATTTGAACAATTGATCTGCACAATGTTGACAAC CCACTTGAAGGAAATAATTGATATAACAAGGTCTGGAACCCAGGATTATGAAAGTTTATTACTCACTCCTTACAAT TTAGGTCTTCTTGGTAAAATCAGTACGATAGTGAGATTATTAACAGAAAGGATTCTAAATCATACTATCAGGAATT GGTTGATCCTCCCACCTTCGCTCCGGATGATCGTGAAGCAGGACTTGGAATTCGGCATATTCAGGATTACTTCCAT CCTCAATTCTGATCGGTTCCTGAAGCTTTCTCCAAATAGGAAATACTTGATTGCACAATTAACTGCAGGCTACATT AGGAAATTGATTGAGGGGGATTGCAATATCGATCTAACCAGACCTATCCAAAAGCAAATCTGGAAAGCATTAGGTT GTGTAGTCTATTGTCACGATCCAATGGATCAAAGGGAGTCAACAGAGTTTATTGATATAAATATTAATGAAGAAAT AGACCGCGGGATCGATGGCGAGGAAATCTAAACATATCAAGAATCAGAATTAGTTTAAGAAAAAAGAAGAGGATTA ATCTTGGTTTTCCCCTTGGT

[0122] The underlined sequence is the PIV5 genomic nucleic acid sequence. The italicized and underlined sequences are the entire OspA nucleic acid sequence. The double-underlined sequence is AOS-modified NTTM-OspA. BPBPk Nucleic acid sequence.

[0123] d. pMHD36- PIV5-NTTM-OspA B31

[0124] This article provides pMHD36-PIV5-NTTM-OspA B31 The nucleic acid sequences from 5' to 3' are as follows:

[0125] ACCAAGGGGAAAATGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCTA TGGCCTTCGTGACCGACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCGACA CAAAAATGTCATCCGTGCTTAAAGCATATGAGCGATTCACGCTCACTCAAGAACTGCAAGATCAGAGTGAGGAAGGT ACAATCCCACCTACAACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGATCCCG GCTTCTTCTATTCTGCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACTCACAA TGTTTTCGCTACCATCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATCGAAAGG GTAGAGATCGATGGCTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCTCGTTCAGGTATGAGCCGTGGAGAGAT CAATGCCTATGCTGCACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTCGTTGATTCCGAAGTCG AGGGAACTGCATGGGATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGGATAGTGACT TGCAAGTGCATGACTGCGCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAATATCGTCAGCAAGGCAG GATCAACCCGAGATATCTCCTGCAACCGGAGGCTCGACGAATAATCCAGAATGTAATCCGGAAGGGAATGGTGGTCA GACATTTCCTCACCTTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTATGCTATGGTAGGG GATGTTGGAAAGTATATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAATATGCATTAGGAACTAGATG GCCCACACTTGCTTTAGCTGCATTTTCAGGAGAGCTAACAAAGCTAAAGTCCCTCATGGCATTATACCAGACCCTTG GTGAGCAGGCCCGATATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTTGCTGCAGCAAACTACCCACTGCTA TATAGCTATGCTATGGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTCTCCAGATCATACATGAA CAAGACATATTTCCAATTGGGAATGGAAACTGCAAGAAAACAACAGGGTGCAGTTGACATGAGGATGGCAGAAGATC TCGGTCTAACTCAAGCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACAGCAAATCGAGGGGCAGAC ACCAGGGGAGGAGTCAACCCGTTCTCATCTGTCACTGGGACAACTCAGGTGCCCGCTGCAGCAACAGGTGACACACT CGAGAGTTACATGGCAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCATGATGATGAGATGCCACCAT TGGAAGAGGAGGAAGAGGACGACACATCTGCAGGTCCACGCACTGGACCAACTCTTGAACAAGTGGCCTTGGACATC CAGAACGCAGCAGTTGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGGGTGATCTTGACATCTAGACAAT TCAGATCCCAATCTAAAATTGACATACCTAATTGATTAGTTAGATGGAACTACAGTGGATTCCATAAGGTTCCTGCC TACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGGTGCCAACAGCGCAATCCA CAATCTACAATGGATCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATAAGCTCATAGAGACAGGCCTGAATAC TGTAGAGTATTTTACTTCCCAACAAGTCACAGGAACATCCTCTCTTGGAAAGAATACAATACCACCAGGGGTCACAG GACTACTAACCAATGCTGCAGAGGCAAAGATCCAAGAGTCAACTAACCATCAGAAGGGCTCAGTTGGTGGGGGTGCA AAACCAAAGAAACCGCGACCAAAAATTGCCATTGTGCCAGCAGATGACAAAACAGTGCCCGGAAAGCCGATCCCAAA CCCTCTATTAGGTCTGGACTCCACCCCGAGCACCCAAACTGTGCTTGATCTAAGTGGGAAAACATTACCATCAGGAT CCTATAAGGGGGTTAAGCTTGCGAAATTTGGAAAAGAAAATCTGATGACACGGTTCATCGAGGAACCCAGAGAGAAT CCTATCGCAACCAGTTCCCCCATCGATTTTAAGAGGGGCAGGGATACCGGCGGGTTCCATAGAAGGGAGTACTCAAT CGGATGGGTGGGAGATGAAGTCAAGGTCACTGAGTGGTGCAATCCATCCTGTTCTCCAATCACCGCTGCAGCAAGGC GATTTGAATGCACTTGTCACCAGTGTCCAGTCACTTGCTCTGAATGTGAACGAGATACTTAATACAGTGAGAAATTT GGACTCTCGGATGAATCAACTGGAGACAAAAGTAGATCGCATTCTCTCATCTCAGTCTCTAATCCAGACCATCAAGA ATGACATAGTTGGACTTAAAGCAGGGATGGCTACTTTAGAAGGAATGATTACAACTGTGAAAATCATGGACCCGGGA GTTCCCAGTAATGTTACTGTGGAAGATGTACGCAAGACACTAAGTAACCATGCTGTTGTTGTGCCAGAATCATTCAA TGATAGTTTCTTGACTCAATCTGAAGATGTAATTTCACTTGATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAGA AGATTGTCAGGAAGGTTCCTCCTCAGAAGGATCTGACTGGATTGAAGATTACACTAGAGCAATTGGCAAAGGATTGC ATCAGCAAACCGAAGATGAGGGAAGAGTATCTCCTCAAAATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCT CAAGAAAGCAATCATCCGCAGTGCAATTTGATCAAGAAACACCCAATTACACTACACTGGTATGACACTGTACTAAC CCTGAGGGTTTTAGAAAAAACGATTAACGATAAATAAGCCCGAACACTACACACTACCTGAGGCAGCCATGCCATCC ATCAGCATTCCCGCAGACCCCACCAATCCACGTCAATCAATAAAAGCGTTCCCAATTGTGATCAACAGTGATGGGGG TGAGAAAGGCCGCTTGGTTAAACAACTACGCACAACCTACTTGAATGACCTAGATACTCATGAGCCACTGGTGACAT TCATAAATACCTATGGATTCATCTACGAACAGGATCGGGGGAATACCATTGTCGGAGAGGATCAACTTGGGAAGAAA AGAGAGGCTGTGACCGCTGCAATGGTTACCCTTGGATGTGGGCCTAATCTACCATCATTAGGGAATGTCCTGGGACA ACTGAGGGAATTCCAGGTCACTGTTAGGAAGACATCCAGCAAAGCGGAAGAGATGGTCTTTGAAATTGTTAAGTATC CGAGAATATTTCGGGGTCATACATTAATCCAGAAAGGACTAGTCTGTGTCTCCGCAGAAAAATTTGTTAAGTCACCA GGGAAAATACAATCTGGAATGGACTATCTCTTCATTCCGACATTTCTGTCAGTGACTTACTGTCCAGCTGCAATCAA ATTCAGGTACCTGGCCCCATGTTGAAAATGAGATCAAGATACACTCAGAGCTTACAACTTGAACTAATGATAAGAA TCCTGTGTAAGCCCGATTCGCCACTTATGAAGGTCCATACCCCTGACAAGGAGGGAAGAGGATGTCTTGTATCAGTA TGGCTGCATGTATGCAACATCTTCAAATCAGGAAACAAGAATGGCAGTGAGTGGCAGGAATACTGGATGAGAAAGTG TGCTAACATGCAACTTGAAGTGTCGATTGCAGATATGTGGGGACCAACTATCATAATTCATGCCAGAGGTCACATTC CCAAAAGTGCTAAGTTGTTTTTTGGAAAGGGTGGATGGAGCTGCCATCCACTTCACGAAGTTGTTCCAAGTGTCACT AAAACACTATGGTCCGTGGGCTGTGAGATTACAAAGGCGAAGGCAATAATACAAGAGAGTAGCATCTCTCTTCTCGT GGAGACTACTGACATCATAAGTCCAAAAGTCAAAATTTCATCTAAGCATCGCCGCTTTGGGAAATCAAATTGGGGTC TGTTCAAGAAAACTAAATCACTGCCTAACCTGACGGAGCTGGAATGACTGACCTCTAATCGAGACTACACCGCCGCA AACTATAGGTGGGTGGTACCTCAGTGATTAATCTTGTAAGCACTGATCGTAGGCTACAACACACTAATATTATCCAG ATTAGAGAGCTTAATTAGCTCTGTATTAATAATAACACTACTATTCCAATAACTGGAATCACCAGCTTGATTTATCT CCAAAATGATTCAAAGAAAACAAATCATATTAAGACTATCCTAAGCACGAACCCATATCGTCCTTCAAATCATGGGT ACTATAATTCAATTTCTGGTGGTCTCCTGTCTATTGGCAGGAGCAGGCAGCCTTGATCCAGCAGCCCTCATGCAAAT CGGTGTCATTCCAACAAATGTCCGGCAACTTATGTATTATACTGAAGCTTCATCAGCATTCATTGTTGTGAAGTTAA TGCCTACAATTGACTCGCCGATTAGTGGATGTAATATAACATCAATTTCAAGCTATAATGCAACAGTGACAAAACTC CTACAGCCGATCGGTGAGAATTTGGAGACGATTAGGAACCAGTTGATTCCAACTCGGAGGAGACGCCGGTTTGCAGG GGTGGTGATTGGATTAGCTGCATTAGGAGTAGCTACTGCCGCACAGGTCACTGCCGCAGTGGCACTAGTAAAGGCAA ATGAAAATGCTGCGGCTATACTCAATCTCAAAAATGCAATCCAAAAAACAAATGCAGCAGTTGCAGATGTGGTCCAG GCCACACAATCACTAGGAACGGCAGTTCAAGCAGTTCAAGATCACATAAACAGTGTGGTAAGTCCAGCAATTACAGC AGCCAATTGTAAGGCCCAAGATGCTATCATTGGCTCAATCCTCAATCTCTATTTGACCGAGTTGACAACCATCTTCC ACAATCAAATTACAAACCCTGCATTGAGTCCCATTACAATTCAAGCTTTAAGGATCCTACTGGGGAGTACCTTGCCG ACTGTGGTCGAAAAATCTTTCAATACCCAGATAAGTGCAGCTGAGCTTCTCTCATCAGGGTTATTGACAGGCCAGAT TGTGGGATTAGATTTGACCTATATGCAGATGGTCATAAAAATTGAGCTGCCAACTTTAACTGTACAACCTGCAACCC AGATCATAGATCTGGCCACCATTTCTGCATTCATTAACAATCAAGAAGTCATGGCCCAATTACCAACACGTGTTATG GTGACTGGCAGCTTGATCCAAGCCTATCCCGCATCGCAATGCACCATTACACCCAACACTGTGTACTGTAGGTATAA TGATGCCCAAGTACTCTCAGATGATACTATGGCTTGCCTCCAAGGTAACTTGACAAGATGCACCTTCTCTCCAGTGG TTGGGAGCTTTCTCACTCGATTCGTGCTGTTCGATGGAATAGTTTATGCAAATTGCAGGTCGATGTTGTGCAAGTGC ATGCAACCTGCTGCTGTGATCCTACAGCCGAGTTCATCCCCTGTAACTGTCATTGACATGTACAAATGTGTGAGTCT GCAGCTTGACAATTCTCAGATTCACCATCACTCAATTGGCCAATGTAACCTACAATAGCACCATCAAGCTTGAATCAT CCCAGATCTTGTCTATTGATCCGTTGGATATATCCCAGAATCTAGCTGCGGTGAATAAGAGTCTAAGTGATGCACTA CAACACTTAGCACAAAGTGACACATATCTTTCTGCAATCACATCAGCTACGACTACAAGTGTATTATCCATAATAGC AATCTGTCTTGGATCGTTAGGTTTAATATTAATAATCTTGCTCAGTGTAGTTGTGTGGAAGTTATTGACCATTGTCG TTGCTAATCGAAATAGAATGGAGAATTTTGTTTATCATAAATAAGCATTCCACCACTCACGATCTGATCTCAGTGAG AAAAAATCAACCTGCAACTCTTGGAACAAGATAAGACAGTCCATCCATTAGTAATTTTTAAGAAAAAAACGATAGGACC GAACCTAGTATTGAAAGAACCGTCTCGGTCCAATCTAGGTAATCGAGCTGATACCGTCTCGGAAAGCTCCAAATCATGC TGCCTGATCCGGAAGATCCGGAAAGCAAGAAAGCTACAAGGAGAGCAGGAAACCTAATTATCTGCTTCCTATTCATC TTCTTTCTGTTTGTAACCTTCATTGTTCCAACTCTAAGACACTTGCTGTCCTAAACACCTGCTATAGGCTATCCACTG CATCATCTTACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGAC

[0126]

[0127] GCCTGATCCTGGCCCTGATCGCCTGCAAGCAGAACGTGAGCTCCCTGGACGAGAAATAGCGTGTCC GTGGATCTGCCCGGCGAGATGAAGGTGCTGGTGAGCAAGGGAAACAAGGACGGCAAGTATGATCTGATCGCCA CCGTGGACAAGCTGGAGCTGAAGGGCACATCCGATAAGAACAATGGATCTGGCGTGCTGGAGGGAGTGAAGGCAGA CAAGAGCAAGGTGAAGCTGACCATCTCCGACGATCTGGGCCAGACCACACTGGAGGTGTTCAAGGAGGACGGCAAG ACACTGGTGTCTAAGAAGGTGACCAGCAAGGATAAGTCTAGCACAGAGGAGAGTTTAATGAGAAGGGCGAGGTGT CCGAGAAGATCATCACCCGGGCCGACGGCCAAGACTGGAGTACACCGGCATCAAGTCCGATGGCTGCTGGCAAGGC CAAGGAGGTGCTGAAGGGCTATGTGCTGGAGGGCACCCTGACAGCCGAGAAGACCACACTGGTGGTGAAGGAGGGC ACCGTGACACTGTCCAAGAACATCTCTAAGAGCGGCGAGGTGTCTGTGGAGCTGAATGACACCGATTCCTCTGCCG CCACCAAGAAGACAGCCGCCTGGAACAGCGGCACCTCCACACTGACCATCACAGTGAATAGCAAGAAGACAAAGGA CCTGGTGTTCACCAAGGAGAACACCATCACAGTGCAGCAGTACGATTCTAATGGAACCAAGCTGGAGGGAAGCGCC GTGGAGATCACAAAGCTGGATGAGATCAAGAACGCCCTGAAGTGAACGCGTGCTAGCTCTCCTGCCATACTTCCTA CTCACATCATATCTATTTTAAAGAAAAAATAGGCCCGAACACTAATCGTGCCGGCAGTGCCACTGCACACACAACA CTACACATACAATACACTACAATGGTTGCAGAAGATGCCCCTGTTAGGGCCACTTGCCGAGTATTATTTCGAACAA CAACTTTAATCTTTCTATGCACACTACTAGCATTAAGCATCTCTATCCTTTATGAGAGTTTAATAACCCAAAAGCA AATCATGAGCCAAGCAGGCTCAACTGGATCTAATTCTGGATTAGGAAGTATCACTGATCTTCTTAATAATATTCTC TCTGTCGCAAATCAGATTATATATAACTCTGCAGTCGCTCTACCTCTACAATTGGACACTCTTGAATCAACACTCC TTACAGCCATTAAGTCTCTTCAAACCAGTGACAAGCTAGAACAGAACTGCTCGTGGAGTGCTGCACTGATTAATGA TAATAGATACATTAATGGCATCAATCAGTTCTATTTTTCAATTGCTGAGGGTCGCAATCTGACACTTGGCCCACTT CTTAATATGCCTAGTTTCATTCCAACTGCCACGACACCAGAGGGCTGCACCAGGATCCCATCATTCTCGCTCACTA AGACACACTGGTGTTATACACACAATGTTATCCTGAATGGATGCCAGGATCATGTATCCTCAAATCAATTTGTTTC TATGGGAATCATTGAACCCACTTCTGCCGGGTTTCCATTCTTTCGAACCCTAAAGACTCTATATCTCAGCGATGGG GTCAATCGTAAGAGCTGCTCTATCAGTACAGTTCCGGGGGGTTGTATGATGTACTGTTTTGTTTCTACTCAACCAG AGAGGGATGACTACTTTTCTGCCGCTCCTCCAGAACAACGAATTATTATAATGTACTATAATGATACAATCGTGGA GCGCATAATTAATCCACCCGGGGTACTAGATGTATGGGCAACATTGAACCCAGGAACAGGAAGCGGGGTATATTAT TTAGGTTGGGTGCTCTTTCCAATATATGGCGGCGTGATTAAAGGTACGAGTTTATGGAATAATCAAGCAAATAAAT ACTTTATCCCCCAGATGGTTGCTGCTCTCTGCTCACAAAACCAGGCAACTCAAGTCCAAAATGCTAAGTCATCATA CTATAGCAGCTGGTTTGGCAATCGAATGATTCAGTCTGGGATCCTGGCATGTCCTCTTCGACAGGATCTAACCAAT GAGTGTTTAGTTCTGCCCTTTTCTAATGATCAGGTGCTTATGGGTGCTGAAGGGAGATTATACATGTATGGTGACT CGGTGTATTACTATCAAAGAAGCAATAGTTGGTGGCCTATGACCATGCTGTATAAGGTAACCATAACATTCACTAA TGGTCAGCCATCTGCTATATCAGCTCAGAATGTGCCCACACAGCAGGTCCCTAGACCTGGGACAGGAGACTGCTCT GCAACCAATAGATGTCCCGGTTTTTGCTTGACAGGAGTGTATGCCGATGCCTGGTTACTGACCAACCCTTCGTCTA CCAGTACATTTGGATCAGAAGCAACCTTCACTGGTTCTTATCTCAACACAGCAACTCAGCGTATCAATCCGACGAT GTATATCGCGAACAACACACAGATCATAAGCTCACAGCAATTTGGATCAAGCGGTCAAGAAGCAGCATATGGCCAC ACAACTTGTTTTAGGGACACAGGCTCTGTTATGGTATACTGTATCTATATTATTGAATTGTCCTCATCTCTCTTAG GACAATTTCAGATTGTCCCATTTATCCGTCAGGTGACACTATCCTAAAGGCAGAAGCCTTCAGGTCTGACCCAGCC AATCAAAGCATTATACCAGACCATGGAATGCATACCAAACATTATTGACACTAATGACACACAAAATTGGTTTTAA GAAAAACCAAGAGAACAATAGGCCAGAATGGCTGGGTCTCGGGAGATATTACTCCCTGAAGTCCATCTCAATTCAC CAATTGTAAAGCATAAGCTATACTATTACATTCTACTTGGAAACCTCCCAAATGAGATCGACCTTGACGATTTAGG TCCATTACATAATCAAAATTGGAATCAGATAGCACATGAAGAGTCTAACTTAGCTCAACGCTTGGTAAATGTAAGA AATTTTCTAATTACCCACATCCCTGATCTTAGAAAGGGCCATTGGCAAGAGTATGTCAATGTAATACTGTGGCCGC GAATTCTTCCCTTGATCCCGGATTTTAAAATCAATGACCAATTGCCTCTGCTCAAAAATTGGGACAAGTTAGTTAA AGAATCATGTTCAGTAATCAATGCAGGTACTTCCCAGTGCATTCAGAATCTCAGCTATGGACTGACAGGTCGTGGG AACCTCTTTACACGATCACGTGAACTCTCTGGTGACCGCAGGGATATTGATCTTAAGACAGTTGTGGCAGCATGGC ATGACTCAGACTGGAAAAGAATAAGTGATTTTTGGATTATGATCAAATTCCAGATGAGACAATTAATTGTTAGGCA AACAGATCATAATGATTCTGATTTAATCACGTATATCGAAAATAGAGAAGGCATAATCATCATAACCCCTGAACTG GTAGCATTATTTAACACTGAGAATCATACACTAACATACATGACCTTTGAAATTGTACTGATGGTTTCAGATATGT ACGAAGGTCGTCACAACATTTTATCACTATGCACAGTTAGCACTTACCTGAATCCTCTGAAGAAAAGAATAACATA TTTATTGAGCCTTGTAGATAACTTAGCTTTTCAGATAGGTGATGCTGTATATAACATAATTGCTTTGCTAGAATCC TTTGTATATGCACAGTTGCAAATGTCAGATCCCATCCCAGAACTCAGAGGACAATTCCATGCATTCGTATGTTCTG AGATTCTTGATGCACTAAGAGGAACTAATAGTTTCACCCAGGATGAATTAAGAACTGTGACAACTAATTTGATATC CCCATTCCAAGATCTGACCCCAGATCTTACGGCTGAATTGCTCTGTATAATGAGGCTTTGGGGACACCCCATGCTC ACTGCCAGTCAAGCTGCAGGAAAGGTACGCGAGTTCTATGTGTGCTGGAAAAGTATTAGACTTTCCCACCATTATGA AAACACTAGCCTTTTTCCATACTATTCTGATCAATGGATACAGGAGGAAGCATCATGGAGTATGGCCACCCTTAAA CTTACCGGGTAATGCTTCAAAGGGTCTCACGGAACTTATGAATGACAATACTGAAATAAGCTATGAATTCACACTT AAGCATTGGAAGGAAGTCTCTCTTATAAAATTCAAGAAATGTTTTGATGCAGACGCAGGTGAGGAACTCAGTATAT TTATGAAAGATAAGGCAATTAGTGCCCCAAAACAAGACTGGATGAGTGTGTTTAGAAGAAGCCTAATCAAACAGCG CCATCAGCATCATCAGGTCCCCCTACCAAATCCATTCAATCGACGGCTGTTGCTAAACTTTCTCGGAGATGACAAA TTCGACCCGAATGTGGAGCTACAGTATGTAACATCAGGTGAGTATCTACATGATGACACGTTTTGTGCATCATATT CACTAAAAGAGAAGGAAATTAAACCTGATGGTCGAATTTTTGCAAAGTTGACTAAGAGAATGAGATCATGTCAAGT TATAGCAGAATCTCTTTTAGCGAACCATGCTGGGAAGTTAATGAAAGAGAATGGTGTTGTGATGAATCAGCTATCA TTAACAAAATCACTATTAACAATGAGTCAGATTGGAATAATATCCGAGAAAGCTAGAAAGTCAACTCGAGATAACA TAAATCAACCTGGTTTCCAGAATATCCAGAGAAATAAATCACATCACTCCAAGCAAGTCAATCAGCGAGATCCAAG TGATGACTTTGAATTGGCAGCATCTTTTTTAACTACTGATCTCAAAATATTGTTTACAATGGAGGTACCAGACA ATTATCCCATTTGCTCAATCATTAAACAGAATGTATGGTTATTCCTCATCTCTTTGAGTGGATTCACTTACGGCTAA TGCGTAGTACACTTTACGTGGGGGATCCCTTCAACCCACCAGCAGATACCAGTCAATTTGATCTAGATAAAGTAAT TAATGGAGATATCTTCATTGTATCACCCAGAGGTGGAATTGAAGGGCTGTGTCAAAAGGCTTGGACAATGATATCT ATCGCTGTGATAATTCTATCTGCCACAGAGTCTGGCACACGAGTAATGAGTATGGTGCAGGGAGATAATCAAGCAA TTGCTGTCACCACACGAGTACCAAGGAGCCTGCCGACTCTTGAGAAAAAGACTATTGCTTTTTAGATCTTGTAATCT ATTCTTTGAGAGGTTAAAATGTAATAATTTTGGATTAGGTCACCATTTGAAAGAACAAGAGACTATCATTAGTTCT CACTTCTTTGTTTATAGCAAGAGAATATTCTATCAGGGGAGGATTCTAACGCAAGCCTTAAAAAATGCTAGTAAGC TCTGCTTGACAGCTGATGTCCTAGGAGAATGCACCCAATCATCATGTTCTAATCTTGCAACTACTGTCATGAGGTT AACTGAGAATGGTGTTGAAAAAGATATCTGTTTCTACTTGAATATCTATATGACCATCAAACAGCTCTCCTATGAT ATCATCTTCCCTCAAGTGTCAATTCCTGGAGATCAGATCACATTAGAATACATAAATAATCCACACCTGGTATCAC GATTGGCTCTTTTGCCATCCCAGTTAGGAGGTCTAAACTACCTGTCATGCAGTAGGCTGTTCAATCGAAACATAGG CGACCCGGTGGTTTCCGCAGTTGCAGATCTTAAGAGATTAATTAAATCAGGATGTATGGATTACTGGATCCTTTAT AACTTATTAGGGAGAAAACCGGGAAACGGCTCATGGGCTACTTTAGCAGCTGACCCGTACTCAATCAATATAGAGT ATCAATACCCTCCAACTACAGCTCTTAAGAGGCACACCCAACAAGCTCTGATGGAACTCAGTACGAATCCAATGTT ACGTGGCATATTCTCTGACAATGCACAGGCAGAAGAAAATAACCTTGCTAGGTTTCTCCTGGATAGGGAGGTGATC TTTCCGCGTGTAGCTCACATCATCATTGAGCAAACCAGTGTCGGGAGGAGAAAACAGATTCAAGGATATTTGGATT CAACTAGATCGATAATGAGGAAATCACTAGAAATTAAGCCCTTATCCAATAGGAAGCTTAATGAAATACTGGATTA CAACATCAATTACCTAGCTTACAATTTGGCATTACTCAAGAATGCTATTGAACCTCCGACTTATTTGAAGGCAATG ACACTTGAAACATGTAGCATCGACATTGCAAGGAACCTCCGGAAGCTCTCCTGGGCCCCACTCTTGGGTGGGAGAA ATCTTGAAGGATTAGAGACGCCAGATCCCATTGAAATTACTGCAGGAGCATTAATTGTTGGATCGGGCTACTGTGA ACAGTGTGCTGCAGGAGACAATCGATTCACATGGTTTTTCTTGCCATCTGGTATCGAGATAGGAGGGGATCCCCGT GATAATCCTCCTATCCGTGTACCGTACATTGGCTCCAGGACTGATGAGAGGAGGGTAGCCTCAATGGCATACATCA GGGGTGCCTCGAGTAGCTTAAAAGCAGTTCTTAGACTGGCGGGAGTGTACATCTGGGCATTCGGAGATACTCTGGA GAATTGGATAGATGCACTGGATTTGTCTCACACTAGAGTTAACATCACACTTGAACAGCTGCAATCCCTCACCCCA CTTCCAACCTCTGCCAATCTAACCCATCGGTTGGATGATGGCACAACTACCCTAAAGTTTACTCCTGCGAGCTCTT ACACCTTTTCAAGTTTCACTCATATATCAAATGATGAGCAATACCTGACAATTAATGACAAAACTGCAGATTCAAA TATAATCTACCAACAGTTAATGATCACTGGACTCGGAATCTTAGAAACATGGAATAATCCCCCAATCAATAGAACA TTCGAAGAATCTACCCTACATTTGCACACTGGTGCATCATGTTGTGTCCGACCTGTGGACTCCTGCATTCTCTCAG AAGCATTAACAGTCAAGCCACATATTACAGTACCGTACAGCAATAAATTTGTATTTGATGAGGACCCGCTATCTGA ATATGAAACTGCAAAACTGGAATCGTTATCATTCCAAGCCCAATTAGGCAACATTGATGCTGTAGATATGACAGGT AAATTAACATTATTGTCCCAATTCACTGCAAGGCAGATTATCAATGCAATCACTGGACTCGATGAGTCTGTCTCTC TTACTAATGATGCCATTGTTGCATCAGACTATGTCTCCAATTGGATTAGTGAATGCATGTATACCAAATTAGATGA ATTATTTATGTATTGTGGGTGGGAACTACTATTGGAACTATCCTATCAAATGTATTATCTGAGGGTAGTTGGGTGG AGTAATATAGTGGATTATTCTTACATGATCTTGAGAAGAATCCCGGGTGCAGCATTAAACAATCTGGCATCTACAT TAAGTCATCCAAAACTTTTCCGACGAGCTATCAACCTAGATATAGTTGCCCCCTTAAATGCTCCTCATTTTGCATC TCTGGACTACATCAAGATGAGTGTGGATGCAATACTCTGGGGCTGTAAAAGAGTCATCAATGTGCTCTCCAATGGA GGGGACTTAGAATTAGTTGTGACATCTGAAGATAGCCTTATTCTCAGTGACCGATCCATGAATCTCATTGCAAGGA AATTAACTTTATTCACTGATTCACCATAATGGTTTGGAACTACCAAAGATTAAGGGTTCTCTCCTGATGAGAA GTGTTTCGCTTTGACAGAATTTGAGGAAAGTGGTGAACTCAGGGTTGAGTTCAATAGAGAACCTATCAAATTTT ATGTACAATGTGGAGAACCCACGGCTTGCAGCATTCGCCAGCAACAATTACTACCTGACCAGAAAATTATTGAATT CAATACGAGATACTGAGTCGGGTCAAGTAGCAGTCCACCTCATATTATGAATCATTAGAATATTGATAGTCTTAA GCTAACCCCACATGTGCCTGGCACCTCATGCATTGAGGATGATAGTCTATGTACAAATGATTACATAATCTGGATC ATAGAGTCTAATGCAAACTTGGAGAAGTATCCAATTCCAAATAGCCCTGAGGATGATTCCAATTTCCATAACTTTA AGTTGAATGCTCCATCGCACCATACCTTACGCCCATTAGGGTTGTCATCAACTGCTTGGTATAAGGGTATAAGCTG CTGCAGGTACCTTGAGCGATTAAAGCTACCACAAGGTGATCATTTATATTGCAGAAGGTAGTGGTGCCAGTATG ACAATCATAGAATACCTATTCCCAGGAAAAGATATTACAATTCTTTATTTAGTAGTGGTGACAATCCCCCAC AAAGAAATTATGCACCAATGCCTACTCAGTTCATTGAGAGTGTCCCATACAAGCTCTGGCAAGCACACACAGATCA ATATCCCGAGATTTGAGGACTTCATCCCTCTATGGAACGGAAACGCCGCCATGACTGACATAGGAATGACAGCT TGTGTAGAATTCATCATCAATCGAGTCGGCCCAAGGACTTGCAGTTTAGTACATGTAGATTTGGAATCAAGTGCAA GCTTAAATCAACAATGCCTGTCAAAGCCGATAATTAATGCTATCATCACTGCTACAACTGTTTTGTGCCCTCATGG GGTGCTTATTCTGAAATATAGTTGGTTGCCATTTACTAGATTTAGTACTTTGATCACTTTCTTATGGTGCTACTTT GAGAGAATCACTGTTCTTAGGAGCACATATTCTGATCCAGCTAATCATGAGGTTTATTTAATTTGTATCCTTGCCA ACAACTTTGCATTCCAGACTGTCTCGCAGGCAACAGGAATGGCGATGACTTTAACTGATCAAGGGTTTACTTTGAT ATCACCTGAAAGAATAAATCAGTATTGGGATGGTCACTTGAAGCAAGAACGTATCGTAGCAGAAGCAATTGATAAG GTGGTTCTAGGAGAAAATGCTCTATTTAATTCGAGTGATAATGAATTAATTCTCAAATGTGGAGGGACACCAAATG CACGGAATCTCATCGATATCGAGCCAGTCGCAACTTTCATAGAATTTGAACAATTGATCTGCACAATGTTGACAAC CCACTTGAAGGAAATAATTGATATAACAAGGTCTGGAACCCAGGATTATGAAAGTTTATTACTCACTCCTTACAAT TTAGGTCTTCTTGGTAAAATCAGTACGATAGTGAGATTATTAACAGAAAGGATTCTAAATCATACTATCAGGAATT GGTTGATCCTCCCACCTTCGCTCCGGATGATCGTGAAGCAGGACTTGGAATTCGGCATATTCAGGATTACTTCCAT CCTCAATTCTGATCGGTTCCTGAAGCTTTCTCCAAATAGGAAATACTTGATTGCACAATTAACTGCAGGCTACATT AGGAAATTGATTGAGGGGGATTGCAATATCGATCTAACCAGACCTATCCAAAAGCAAATCTGGAAAGCATTAGGTT GTGTAGTCTATTGTCACGATCCAATGGATCAAAGGGAGTCAACAGAGTTTATTGATATAAATATTAATGAAGAAAT AGACCGCGGGATCGATGGCGAGGAAATCTAAACATATCAAGAATCAGAATTAGTTTAAGAAAAAAGAAGAGGATTA ATCTTGGTTTTCCCCTTGGT

[0128] The underlined sequence is the PIV5 genomic nucleic acid sequence. The italicized and underlined sequences are the entire OspA nucleic acid sequence. The double-underlined sequence is AOS-modified NTTM-OspA. B31 Nucleic acid sequence.

[0129] 2. AVLP vaccine sequence

[0130] a. pMHD30-AVLP-Pal

[0131] The pMHD30-AVLP-Pal nucleic acid sequences provided in this article, from 5' to 3', are as follows:

[0132] ACCAAGGGGAAAATGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCT ATGGCCTTCGTGACCGACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCGA CACAAAAATGTCATCCGTGCTTAAAGCATATGAGCGATTCACGCTCACTCAAGAACTGCAAGATCAGAGTGAGGAA GGTACAATCCCACCTACAACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGAT CCCGGCTTCTTCTATTCTGCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACT CACAATGTTTTCGCTACCATCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATC GAAAGGGTAGAGATCGATGGCTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCACGTTCAGGTATGAGCCGTG GAGAGATCAATGCCTATGCTGCACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTCGTTGATTC CGAAGTCGAGGGAACTGCATGGGATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGG ATAGTGACTTGCAAGTGCATGACTGCGCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAATATCGTC AGCAAGGCAGGATCAACCCGAGATATCTCCTGCAACCGGAGGCTCGACGAATAATCCAGAATGTAATCCGGAAGGG AATGGTGGTCAGACATTTCCTCACCTTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTAT GCTATGGTAGGGGATGTTGGAAAGTATATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAAATATGCAT TAGGAACTAGATGGCCCACACTTGCTTTAGCTGCATTTTCAGGAGAGCTAACAAAGCTAAAGTCCTCATGGCATT ATACCAGACCCTTGGTGAGCAGGCCCGATATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTGCTGCAGCA AACTACCCACTGCTATATAGCTATGCTATGGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTTCT CCAGATCATACATGAACAAGACATATTTCCAATTGGGAATGGAACTGCAAGAAAACAACAGGGTGCAGTTGACAT GAGGATGGCAGAAGATCTCGGTCTAACTCAAGCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACA GCAAATCGAGGGGCAGACACCAGGGGAGGAGTCAACCCGTTCTCATCTGTCACTGGGACAACTCAGGTGCCCGCTG CAGCAACAGGTGACACACTCGAGAGTTACATGGCAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCA TGATGATGAGATGCCACCATTGGAAGAGGAAGAGGACGACACATCTGCAGGTCCACGCACTGGACCAACTCTT GAACAAGTGGCCTTGGACATCCAGAACGCAGCAGTTGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGG GTGATCTTGACATCTAGACAAATTCAGATCCCAATCTAAAATTGACATACCTAATTGATTAGTTAGATGGAACTACA GTGGATTCCATAAGGTTCCTGCCTACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGAC TGCCGGTGCCAACAGCGCAATCCACAATCTACAATGGATCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATA AGCTCATAGAGACAGGCCTGAATACTGTAGGTATTTTACTTCCCAACAAGTCACAGGAACATCCCTCTTTGGAAA GAATACAATACCACCAGGGGTCACAGGACTACTAACCAATGCTGCAGAGGCAAAGATCCAAGAGTCAACTAACCAT CAGAAGGGCTCAGTTGGTGGGGGGTGCAAACCAAAGAAACCGCGACCAAAAATTGCCATTGTGCCAGCAGATGACA AAACAGTGCCCGGAAAGCCGATCCCAAACCCTCTTATTAGGTCTGGACTCCACCCCGAGCACCCAAACTGTGCTTGA TCTAAGTGGGAAACATTACCATCAGGATCCTATAAGGGGGTTAAGCTTGCGAAATTTGGAAAGAAAATCTGATG ACACGGTTCATCGAGGAACCCAGAGAGAATCCTATCGCAACCAGTTCCCCCATCGATTTTAAGAGGGGCAGGGATA CCGGCGGGTTCCATAGAAGGGAGTACTCAATCGGATGGGTGGAGATGAAGTCAAGGTCACTGAGTGGTGCAATCC ATCCTGTTTCTCCAATCACCGCTGCAGCAAGGCGATTTGAATGCACTTGTCACCAGTGTCCAGTCACTTGCTCTGAA TGTGAACGAGATACTTAATACAGTGAGAAATTTGGACTTCCGGATGAATCAACTGGAGACAAAAGTAGATCGCATT CTTCCATCTCAGTCTCTAATCCAGACCATCAAGAATGACATAGTTGGACTTAAAGCAGGGATGGCTACTTTAGAAG GAATGATTACAACTGTGAAAATCATGGACCCGGGAGTTCCCAGTAATGTTACTGTGGAAGATGTACGCAAGACACT AAGTAACCATGCTGTTGTTGTCCAGAATCATTCCAATGATAGTTTCTTGACTCAATCTGAAGATGTAATTTCACTT GATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAAGAAGATTGTCAGGAAGGTTCCTCCTCAGAAGGATCTGACTG GATTGAAGATTACACTAGAGCAATTGGCAAAGGATTGCATCAGCAAACCGAAGATGAGGGAAGAGTATCTCCTCAA AATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCTCAAGAAAGCAATCATCCGCAGTGCAATTTGATCAAGA AACACCCAATTACACTACACTGGTATGACACTGTACTAACCCTGAGGGTTTTAGAAAAAACGATTAACGATAAATA AGCCCGAACACTACACACTACCTGCGGCCGCCACCATGAAATCAGGAGTGAAACTGGACGAGCACGCTAATCAGGG AGACGCAGTCAGCACCCAGCCAAACCCCGAGAATGTCGCACAGGTCACCGTGGACCCACTGAACGATCCCAATTCT CCTCTGGCCAAGCGGAGCGTGTACTTCGACTTTGATAGCTATTCCGTGCAGGACCAGTACCAGGCCCTGCTGCAGC AGCACGCCCAGTATCTGAAGAGCCACCCCCAGAGGCACATCCTGATCCAGGGCAACACCGATGAGCGCGGCACATC CGAGTACAATCTGGCCCTGGGCCAGAAGAGGGCAGAGGCCGTGCGGAGAGCCCTGTCCCTGCTGGGCGTGGGCGAC GCCCAGATGGAGGCCGTGTCTCTGGGCAAGGAGAAGCCCGTGGCCCTGGGGCACGACGAAGCCTCTTGGGCACAGA ATCGCCGAGCAGACCTGGTGTATCAGCAGTGATGAGCTAGCTCTCCTGCCATACTTCCTACTCACATCATATCTAT TTTAAAGAAAAAATAGGCCCGAACACTAATCGTGCCGGCAGTGCCACTGCACACACAACACTACACATACAATACA CTCGCCGGCGCGCCTAGCATGAAAAAGCCTGAACTCACCGCGACGTCTGTCGAGAAGTTTCTGATCGAAAAGTTCG ACAGCGTCTCCGACCTGATGCAGCTCTCGGAGGGCGAAGAATCTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGG ATATGTCCTGCGGGTAAATAGCTGCGCCGATGGTTTCTACAAAGATCGTTATGTTTATCGGCACTTTGCATCGGCC GCGCTCCCGATTCCGGAAGTGCTTGACATTGGGGAATTCAGCGAGAGCCTGACCTATTGCATCTCCCGCCGTGCAC AGGGTGTCACGTTGCAAGACCTGCCTGAAACCGAACTGCCCGCTGTTCTGCAGCCGGTCGCGGAGGCCATGGATGC GATCGCTGCGGCCGATCTTAGCCAGACGAGCGGGTTCGGCCCATTCGGACCGCAAGGAATCGGTCAATACACTACA TGGCGTGATTTCATATGCGCGATTGCTGATCCCCATGTGTATCACTGGCAAACTGTGATGGACGACACCGTCAGTG CGTCCGTCGCGCAGGCTCTCGATGAGCTGATGCTTTGGGCCGAGGACTGCCCCGAAGTCCGGCACCTCGTGCACGC GGATTTCGGCTCCAACAATGTCCTGACGGACAATGGCCGCATAACAGCGGTCATTGACTGGAGCGAGGCGATGTTC GGGGATTCCCAATACGAGGTCGCCAACATCTTCTTCTGGAGGCCGTGGTTGGCTTGTATGGAGCAGCAGACGCGCT ACTTCGAGCGGAGGCATCCGGAGCTTGCAGGATCGCCGCGGCTCCGGGCGTATATGCTCCGCATTGGTCTTGACCA ACTCTATCAGAGCTTGGTTGACGGCAATTTCGATGATGCAGCTTGGGCGCAGGGTCGATGCGACGCAATCGTCCGA TCCGGAGCCGGGACTGTCGGGCGTACACAAATCGCCCGCAGAAGCGCGGCCGTCTGGACCGATGGCTGTGTAGAAG TACTCGCCGATAGTGGAAACCGACGCCCCAGCACTCGTGGGGATCGGGAGATGGGGGAGGCTAACTCTGGTGGCGG TGGCTCGGGCGGAGGTGGGTCGGGTGGCGGCGGATCAATGGCATCCTATCCTTGTCACCAGCACGCGTCTGCATTC GACCAGGCCGCTAGAAGCAGAGGGCACTCCAACCGCAGAACCGCACTGAGACCCCGGAGACAGCAGGAGGCCACAG AAGTGAGGCTGGAGCAGAAGATGCCCACTCTGCTGCGCGTCTACATCGACGGGCCTCACGGAATGGGCAAAACCAC AACTACCCAGCTGCTGGTGGCCCTGGGAAGTAGGGACGATATCGTGTACGTCCCTGAGCCAATGACCTATTGGCAG GTGCTGGGGGCATCAGAAACAATCGCCAACATCTACACAACTCAGCATCGACTGGACCAGGGAGAGATCTCAGCCG GGGATGCAGCTGTGGTCATGACCAGCGCACAGATTACAATGGGCATGCCATATGCAGTGACCGACGCTGTCCTGGC ACCACACATCGGAGGAGAAGCAGGAAGCTCCCATGCACCACCTCCAGCTCTGACCCTGATCTTCGATCGGCACCCT ATTGCAGCCCTGCTGTGCTACCCAGCTGCAAGATATCTGATGGGCTCTATGACCCCACAGGCCGTGCTGGCTTTTG TCGCACTGATCCCCCCTACACTGCCTGGAACTAATATTGTGCTGGGCGCTCTGCCAGAGGACCGCCATATCGATCG ACTGGCAAAGCGACAGCGACCAGGAGAAAGGCTGGACCTGGCAATGCTGGCCGCTATTAGGCGCGTGTACGGCCTG CTGGCTAACACTGTCCGGTATCTGCAGGGAGGCGGGTCTTGGAGAGAGGACTGGGGGCAGCTGAGTGGAACAGCAG TGCCACCCCAGGGAGCCGAACCTCAGAGCAATGCTGGGCCCCGACCTCACATCGGAGATACTCTGTTCACCCTGTT TCGCGCTCCAGAGCTGCTGGCACCAAACGGCGACCTGTACAACGTGTTCGCATGGGCCCTGGATGTCCTGGCCAAA AGACTGAGGCCCATGCACGTGTTTATTCTGGACTATGATCAGTCCCCTGCAGGATGTCGAGATGCTCTGCTGCAGC TGACTTCCGGCATGGTGCAGACCCATGTCACCACACCCGGGTCTATTCCAACTATTTGTGACCTGGCAAGGACTTT CGCAAGAGAGATGGGAGAGGCTAACAGTGGAGGTGGAGGCAGCGGAGGCGGAGGTTCTGGTGGAGGCGGAAGTATG GTCTTCACACTCGAAGATTTCGTTGGGGACTGGCGACAGACAGCCGGCTACAACCTGGACCAAGTCCTTGAACAGG GAGGTGTGTCCAGTTTGTTTCAGAATCTCGGGGTGTCCGTAACTCCGATCCAAAGGATTGTCCTGAGCGGTGAAAA TGGGCTGAAGATCGACATCCATGTCATCATCCCGTATGAAGGTCTGAGCGGCGACCAAATGGGCCAGATCGAAAAA ATTTTTAAGGTGGTGTACCCTGTGGATGATCATCACTTTAAGGTGATCCTGCACTATGGCACACTGGTAATCGACG GGGTTACGCCGAACATGATCGACTATTTCGGACGGCCGTATGAAGGCATCGCCGTGTTCGACGGCAAAAAGATCAC TGTAACAGGGACCCTGTGGAACGGCAACAAAATTATCGACGAGCGCCTGATCAACCCCGACGGCTCCCTGCTGTTC CGAGTAACCATCAACGGAGTGACCGGCTGGCGGCTGTGCGAACGCATTCTGGCGTAATGAACTAGTGGCAGAAGCC TTCAGGTCTGACCCAGCCAATCAAAGCATTATACCAGACCATGGAATGCATACCAAACATTATTGACACTAATGAC ACACAAAATTGGTTTTAAGAAAAACCAAGAGAACAATAGGCCAGAATGGCTGGGTCTCGGGAGATATTACTCCCTG AAGTCCATCTCAATTCACCAATTGTAAAGCATAAGCTATACTATTACATTCTACTTGGAAACCTCCCAAATGAGAT CGACCTTGACGATTTAGGTCCATTACATAATCAAAATTGGAATCAGATAGCACATGAAGAGTCTAACTTAGCTCAA CGCTTGGTAAATGTAAAAATTTCTAATTACCCACATCCCTGATCTTAGAAAGGGCCATTGGCAAGAGTATGTCA ATGTAATACTGTGGCCGCGAATTCTTCCCTTGATCCCGGATTTTAAATCAATGACCAATTGCCTCTGCTCCAAAAAA TTGGGACAAGTTAGTTAAAAGAATCATGTTCAGTAATCAATGCAGGTACTTCCCAGTGCATTCAGAATCTCAGCTAT GGACTGACAGGTCGTGGGAACCTCTTTACACGATCACGTGAACTCTCTGGTGACCGCAGGGATATTGATCTTAAGA CAGTTGTGGCAGCATGGCATGACTCAGACTGGAAAAAAAGTGATTTTTGGATTATGATCAAATTCCAGATGAG ACAATTAATTGTTAGGCAAACAGATCATAATGATTCTGATTTAATCACGTTATATCGAAAATAGAGAAGGCATAATC ATCATAACCCCTGAACTGGTAGCATTATTTAACACTGAGAATCATACACTAACATACATGACCTTTGAAATTGTAC TGATGGTTTCAGATATGTACGAAGGTCGTCACAACATTTTATCACTATGCACAGTTAGCACTTACCTGAATCCTCT GAAGAAAAGAATAACATATTTATTGAGCCTTGTAGATAACTTAGCTTTTCAGATAGGTGATGCTGTATATAACATA ATTGCTTTGCTAGAATCCTTTGTATATGCACAGTTGCAAATGTCAGATCCATCCCAGAACTCAGAGGACAATTCC ATGCATTCGTATGTTCTGAGATTCTTGATGCACTAAAGAGGAACTAATAGTTTCACCCAGGATGAATTAAGAACTGT GACAACTAATTGATATCCCCATTCCAAGATCTGACCCCAGATCTTACGGCTGAATTGCTCTGTATAATGAGGCTT TGGGGACACCCCATGCTCACTGCCAGTCAAGCTGCAGGAAAGGTACGCGAGTCTATGTGTGCTGGAAAAGTATTAG ACTTTCCCCCATTATGAAAACACTAGCCTTTTTCCATACTATTCTGATCAATGGATACAGGAGGAAGCATCATGG AGTATGGCCACCCTTAAACTTACCGGGTAATGCTTCAAAGGGTCTCACGGAACTTATGAATGACAATACTGAAATA AGCTATGAATTCACACTTAAGCATTGGAAGGAAGTCTCTCTTATAAAATTCAAGAAATGTTTTGATGCAGACGCAG GTGAGGAACTCAGTATATTTATGAAAGATAAGGCAATTAGTGCCCCAAAACAAGACTGGATGAGTTGTTTTTAGAAG AAGCCTAATCAAACAGCGCCATCAGCATCATCAGGTCCCCCTACCAAATCCATTCAATCGACGGCTGTTGCTAAAC TTTCTCGGAGATGACAAATTCGACCCGAATGTGGAGCTACAGTATGTAACATCAGGTGAGTATCTACATGATGACA CGTTTTGTGCATCATATTCACTAAAAGAGAAGGAAATTAAACCTGATGGTCGAATTTTTGCAAAGTTGACTAAGAG AATGAGATCATGTCAAGTTATAGCAGAATCTCTTTTAGCGAACCATGCTGGGAAGTTAATGAAAGAGAATGGTGTT GTGATGAATCAGCTATCATTAACAAAATCACTATTAACAATGAGTCAGATTGGAATAATATCCGAGAAAGCTAGAA AGTCAACTCGAGATAACATAAATCAACCTGGTTTCCAGAATATCCAGAGAAATAAATCACATCACTCCAAGCAAGT CAATCAGCGAGATCCAAGTGATGACTTTGAATTGGCAGCATCTTTTTTAACTACTGATCTCAAAAAATATTGTTTA CAATGGAGGTACCAGACAATTATCCCATTTGCTCAATCATTAAACAGAATGTATGGTTATTCCTCATCTCTTTGAGT GGATTCACTTACGGCTAATGCGTAGTACACTTTACGTGGGGGATCCCTTCAACCCACCAGCAGATACCAGTCAATT TGATCTAGATAAAGTAATTAATGGAGATATCTTCATTGTATCACCCAGAGGTGGAATTGAAGGGCTGTGTCAAAAG GCTTGGACAATGATATCTATCGCTGTGATAATTCTATCTGCCACAGAGTCTGGCACACGAGTAATGAGTATGGTGC AGGGAGATAATCAAGCAATTGCTGTCACCACACGAGTACCAAGGAGCCTGCCGACTCTTGAGAAAAAGACTATTGC TTTTAGATCTTGTAATCTATTCTTTGAGAGGTTAAAATGTAATAATTTTGGATTAGGTCACCATTTGAAAGAACAA GAGACTATCATTAGTTCTCACTTCTTTGTTTATAGCAAGAGAATATTCTATCAGGGGAGGATTCTAACGCAAGCCT TAAAAAATGCTAGTAAGCTCTGCTTGACAGCTGATGTCCTAGGAGAATGCACCCAATCATCATGTTCTAATCTTGC AACTACTGTCATGAGGTTAACTGAGAATGGTGTTGAAAAAGATATCTGTTTCTACTTGAATATCTATATGACCATC AAACAGCTCTCCTATGATATCATCTTCCCTCAAGTGTCAATTCCTGGAGATCAGATCACATTAGAATACATAAATA ATCCACACCTGGTATCACGATTGGCTCTTTTGCCATCCCAGTTAGGAGGTCTAAACTACCTGTCATGCAGTAGGCT GTTCAATCGAAACATAGGCGACCCGGTGGTTTCCGCAGTTGCAGATCTTAAGAGATTAATTAAATCAGGATGTATG GATTACTGGATCCTTTATAACTTATTAGGGAGAAAACCGGGAAACGGCTCATGGGCTACTTTAGCAGCTGACCCGT ACTCAATCAATATAGAGTATCAATACCCTCCAACTACAGCTCTTAAGAGGCACACCCAACAAGCTCTGATGGAACT CAGTACGAATCCAATGTTACGTGGCATATTCTCTGACAATGCACAGGCAGAAGAAAATAACCTTGCTAGGTTTCTC CTGGATAGGGAGGTGATCTTTCCGCGTGTAGCTCACATCATCATTGAGCAAACCAGTGTCGGGAGGAGAAAACAGA TTCAAGGATATTTGGATTCAACTAGATCGATAATGAGGAAATCACTAGAAATTAAGCCCTTATCCAATAGGAAGCT TAATGAAATACTGGATTACAACATCAATTACCTAGCTTACAATTTGGCATTACTCAAGAATGCTATTGAACCTCCG ACTTATTTGAAGGCAATGACACTTGAAACATGTAGCATCGACATTGCAAGGAACCTCCGGAAGCTCTCCTGGGCCC CACTCTTGGGTGGGAGAAATCTTGAAGGATTAGAGACGCCAGATCCCATTGAAATTACTGCAGGAGCATTAATTGT TGGATCGGGCTACTGTGAACAGTGTGCTGCAGGAGACAATCGATTCACATGGTTTTTCTTGCCATCTGGTATCGAG ATAGGAGGGGATCCCCGTGATAATCCTCCTATCCGTGTACCGTACATTGGCTCCAGGACTGATGAGAGGAGGGTAG CCTCAATGGCATACATCAGGGGTGCCTCGAGTAGCTTAAAAGCAGTTCTTAGACTGGCGGGAGTGTACATCTGGGC ATTCGGAGATACTCTGGAGAATTGGATAGATGCACTGGATTTGTCTCACACTAGAGTTAACATCACACTTGAACAG CTGCAATCCCTCACCCCACTTCCAACCTCTGCCAATCTAACCCATCGGTTGGATGATGGCACAACTACCCTAAAGT TTACTCCTGCGAGCTCTTACACCTTTTCAAGTTTCACTCATATATCAAATGATGAGCAATACCTGACAATTAATGA CAAAACTGCAGATTCAAATATAATCTACCAACAGTTAATGATCACTGGACTCGGAATCTTAGAAACATGGAATAAT CCCCCAATCAATAGAACATTCGAAGAATCTACCCTACATTTGCACACTGGTGCATCATGTTGTGTCCGACCTGTGG ACTCCTGCATTCTCTCAGAAGCATTAACAGTCAAGCCACATATTACAGTACCGTACAGCAATAAATTTGTATTTGA TGAGGACCCGCTATCTGAATATGAAACTGCAAAACTGGAATCGTTATTCATTCCAAGCCCAATTAGGCAACATTGAT GCTGTAGATATGACAGGTAAATTAACATTATTGTCCCAATTCACTGCAAGGCAGATTATCAATGCAATCACTGGAC TCGATGAGTCTGTCTCTCTTACTAATGATGCCATTGTTGCATCAGACTATGTCTCCAATTGGATTAGTGAATGCAT GTATACCAAATTAGATGAATTATTTATGTATTGTGGGTGGGAACTACTATTGGAACTATCCTATCAAATGTATTAT CTGAGGGTAGTTGGGTGGAGTAATATAGTGGATTATTCTTACATGATCTTGAGAAGAATCCCGGGTGCAGCATTAA ACAATCTGGCATCTACATTAAGTCATCCAAAACTTTTCCGACGAGCTATCAACCTAGATATAGTTGCCCCCTTAAA TGCTCCTCATTTTGCATCTCTGGACTACATCAAGATGAGTGTGGATGCAATACTCTGGGGCTGTAAAAGAGTCATC AATGTGCTCTCCAATGGAGGGGACTTAGAATTAGTTGTGACATCTGAAGATAGCCTTATTCTCAGTGACCGATCCA TGAATCTCATTGCAAGGAAATTAACTTTATTATCACTGATTCACCATAATGGTTTGGAACTACCAAAGATTAAGGG GTTCTCTCCTGATGAGAAGTGTTTCGCTTTGACAGAATTTTTGAGGAAAGTGGTGAACTCAGGGTTGAGTTCAATA GAGAACCTATTCAAATTTTATGTACAATGTGGAGAACCCACGGCTTGCAGCATTCGCCAGCAACAATTACTACCTGA CCAGAAAATTATTGAATTCAATACGAGATACTGAGTCGGGTCAAGTAGCAGTCACCTCATATTATGAATCATTAGA ATATATTGATAGTCTTAAGCTAACCCCACATGTGCCTGGCACCTCATGCATTGAGGATGATAGTCTATGTACAAAT GATTACATAATCTGGATCATAGAGTCTAATGCAAACTTGGAGAAGTATCCAATTCCAAATAGCCCTGAGGATGATT CCAATTTCCATAACTTTAAGTTGAATGCTCCATCGCACCATACCTTACGCCCATTAGGGTTGTCATCAACTGCTTG GTATAAGGGTATAAGCTGCTGCAGGTACCTTGAGCGATTAAAGCTACCACAAGGTGATCATTTATATATTGCAGAA GGTAGTGGTGCCAGTATGACAATCATAGAATACCTATTCCCAGGAAGAAAGATATATTACAATTCTTTATTTAGTA GTGGTGACAATCCCCCACAAAGAAATTATGCACCAATGCCTACTCAGTTCATTGAGAGTGTCCCATACAAGCTCTG GCAAGCACACACAGATCAATATCCCGAGATTTTTGAGGACTTCATCCCTCTATGGAACGGAAACGCCGCCATGACT GACATAGGAATGACAGCTTGTGTAGAATTCATCATCAATCGAGTCGGCCCAAGGACTTGCAGTTTAGTACATGTAG ATTTGGAATCAAGTGCAAGCTTAAATCAACAATGCCTGTCAAAGCCGATAATTAATGCTATCATCACTGCTACAAC TGTTTTGTGCCCTCATGGGGTGCTTATTCTGAAATATAGTTGGTTGCCATTTACTAGATTTAGTACTTTGATCACT TTCTTATGGTGCTACTTTGAGAGAATCACTGTTCTTAGGAGCACATATTCTGATCCAGCTAATCATGAGGTTTATT TAATTTGTATCCTTGCCAACAACTTTGCATTCCAGACTGTCTCGCAGGCAACAGGAATGGCGATGACTTTAACTGA TCAAGGGTTTACTTTGATATCACCTGAAAGAATAAATCAGTATTGGGATGGTCACTTGAAGCAAGAACGTATCGTA GCAGAAGCAATTGATAAGGTGGTTCTAGGAGAAAATGCTCTATTTAATTCGAGTGATAATGAATTAATTCTCAAAT GTGGAGGGACACCAAATGCACGGAATCTCATCCGATATCGAGCCAGTCGCAACTTTCATAGAATTTGAACAATTGAT CTGCACAATGTTGACAACCCACTTGAAGGAAATAATTGATATAACAAGGTCTGGAACCCAGGATTATGAAAGTTTA TTACTCACTCCTTACAATTTAGGTCTTCTTGGTAAAATCAGTACGATAGTGAGATTATTAACAGAAAGGATTCTAA ATCATACTATCAGGAATTGGTTGATCCTCCCACCTTCGCTCCGGATGATCGTGAAGCAGGACTTGGAATTCGGCAT ATTCAGGATTACTTCCATCCTCAATTCTGATCGGTTCCTGAAGCTTTCTCCAAATAGGAAATACTTGATTGCACAA TTAACTGCAGGCTACATTAGGAAATTGATTGAGGGGGATTGCAATATCGATCTAACCAGACCTATCCAAGCAAA TCTGGAAAGCATTAGGTTGTGTAGTCTATTGTCACGATCCAATGGATCAAAGGGAGTCAACAGAGTTTATTGATAT AAATATTAATGAAGAAATAGACCGCGGGATCGATGGCGAGGAAATCTAAACATATCAAGAATCAGAATTAGTTTAA GAAAAAAGAAGAGGATTAATCTTGGTTTTCCCCTTGGT

[0133] The underlined sequence is the PIV5-AVLP genomic nucleic acid sequence. The italicized and underlined sequences are the entire Pal nucleic acid sequence.

[0134] b. pGOB10-AVLP-NTTM-Pal

[0135] The pGOB10-AVLP-NTTM-Pal nucleic acid sequence provided in this article, from 5' to 3', is as follows:

[0136] ACCAAGGGGAAAATGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCTA TGGCCTTCGTGACCGACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCCGACA CAAAAATGTCATCCGTGCTTAAAGCATATGAGCGATTCACGCTCACTCAAGAACTGCAAGATCAGAGTGAGGAAGGT ACAATCCCACCTACAACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGATCCCG GCTTCTTCTATTCTGCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACTCACAA TGTTTTCGCTACCATCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATCGAAAGG GTAGAGATCGATGGCTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCTCGTTCAGGTATGAGCCGTGGAGAGAT CAATGCCTATGCTGCACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTCGTTGATTCCGAAGTCG AGGGAACTGCATGGGATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGGATAGTGACT TGCAAGTGCATGACTGCGCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAATATCGTCAGCAAGGCAG GATCAACCCGAGATATCTCCTGCAACCGGAGGCTCGACGAATAATCCAGAATGTAATCCGGAAGGGAATGGTGGTCA GACATTTCCTCACCTTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTATGCTATGGTAGGG GATGTTGGAAAGTATATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAATATGCATTAGGAACTAGATG GCCCACACTTGCTTTAGCTGCATTTTCAGGAGAGCTAACAAAGCTAAAGTCCCTCATGGCATTATACCAGACCCTTG GTGAGCAGGCCCGATATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTTGCTGCAGCAAACTACCCACTGCTA TATAGCTATGCTATGGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTCTCCAGATCATACATGAA CAAGACATATTTCCAATTGGGAATGGAAACTGCAAGAAAACAACAGGGTGCAGTTGACATGAGGATGGCAGAAGATC TCGGTCTAACTCAAGCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACAGCAAATCGAGGGGCAGAC ACCAGGGGAGGAGTCAACCCGTTCTCATCTGTCACTGGGACAACTCAGGTGCCCGCTGCAGCAACAGGTGACACACT CGAGAGTTACATGGCAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCATGATGATGAGATGCCACCAT TGGAAGAGGAGGAAGAGGACGACACATCTGCAGGTCCACGCACTGGACCAACTCTTGAACAAGTGGCCTTGGACATC CAGAACGCAGCAGTTGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGGGTGATCTTGACATCTAGACAAT TCAGATCCCAATCTAAAATTGACATACCTAATTGATTAGTTAGATGGAACTACAGTGGATTCCATAAGGTTCCTGCC TACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGGTGCCAACAGCGCAATCCA CAATCTACAATGGATCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATAAGCTCATAGAGACAGGCCTGAATAC TGTAGAGTATTTTACTTCCCAACAAGTCACAGGAACATCCTCTCTTGGAAAGAATACAATACCACCAGGGGTCACAG GACTACTAACCAATGCTGCAGAGGCAAAGATCCAAGAGTCAACTAACCATCAGAAGGGCTCAGTTGGTGGGGGTGCA AAACCAAAGAAACCGCGACCAAAAATTGCCATTGTGCCAGCAGATGACAAAACAGTGCCCGGAAAGCCGATCCCAAA CCCTCTATTAGGTCTGGACTCCACCCCGAGCACCCAAACTGTGCTTGATCTAAGTGGGAAAACATTACCATCAGGAT CCTATAAGGGGGTTAAGCTTGCGAAATTTGGAAAAGAAAATCTGATGACACGGTTCATCGAGGAACCCAGAGAGAAT CCTATCGCAACCAGTTCCCCCATCGATTTTAAGAGGGGCAGGGATACCGGCGGGTTCCATAGAAGGGAGTACTCAAT CGGATGGGTGGGAGATGAAGTCAAGGTCACTGAGTGGTGCAATCCATCCTGTTCTCCAATCACCGCTGCAGCAAGGC GATTTGAATGCACTTGTCACCAGTGTCCAGTCACTTGCTCTGAATGTGAACGAGATACTTAATACAGTGAGAAATTT GGACTCTCGGATGAATCAACTGGAGACAAAAGTAGATCGCATTCTCTCATCTCAGTCTCTAATCCAGACCATCAAGA ATGACATAGTTGGACTTAAAGCAGGGATGGCTACTTTAGAAGGAATGATTACAACTGTGAAAATCATGGACCCGGGA GTTCCCAGTAATGTTACTGTGGAAGATGTACGCAAGACACTAAGTAACCATGCTGTTGTTGTGCCAGAATCATTCAA TGATAGTTTCTTGACTCAATCTGAAGATGTAATTTCACTTGATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAGA AGATTGTCAGGAAGGTTCCTCCTCAGAAGGATCTGACTGGATTGAAGATTACACTAGAGCAATTGGCAAAGGATTGC ATCAGCAAACCGAAGATGAGGGAAGAGTATCTCCTCAAAATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCT CAAGAAAGCAATCATCCGCAGTGCAATTTGATCAAGAAACACCCAATTACACTACACTGGTATGACACTGTACTAAC CCTGAGGGTTTTAGAAAAAACGATTAACGATAAATAAGCCCGAACAC

[0137]

[0138] ACGCAGTCAGCACCCAGCCAAACCCCGAGAATGTCGCACAGGTCACCGTGGACCCACTGAACGATCCC AATTCTCCTCTGGCCAAGCGGAGCGTGTACTTCGACTTTGATAGCTATTCCGTGCAGGACCAGTACCAGGCCCTGC TGCAGCAGCACGCCCAGTATCTGAAGAGCCACCCCCAGAGGCACATCCTGATCCAGGGCAACACCGATGAGCGCGG CACATCCGAGTACAATCTGGCCCTGGGCCAGAAGAGGGCAGAGGCCGTGCGGAGAGCCCTGTCCCTGCTGGGCGTG GGCGACGCCCAGATGGAGGCCGTGTCTCTGGGCAAGGAGAAGCCCGTGGCCCTGGGGCACGACGAAGCCTCTTGGG CACAGAATCGCCGAGCAGACCTGGTGTATCAGCAGTGATAAGCTAGCTCTCCTGCCATACTTCCTACTCACATCAT ATCTATTTTAAAGAAAAAATAGGCCCGAACACTAATCGTGCCGGCAGTGCCACTGCACACACAACACTACACATAC AATACACTCGCCGGCGCGCCTAGCATGAAAAAGCCTGAACTCACCGCGACGTCTGTCGAGAAGTTTCTGATCGAAA AGTTCGACAGCGTCTCCGACCTGATGCAGCTCTCGGAGGGCGAAGAATCTCGTGCTTTCAGCTTCGATGTAGGAGG GCGTGGATATGTCCTGCGGGTAAATAGCTGCGCCGATGGTTTCTACAAAGATCGTTATGTTTATCGGCACTTTGCA TCGGCCGCGCTCCCGATTCCGGAAGTGCTTGACATTGGGGAATTCAGCGAGAGCCTGACCTATTGCATCTCCCGCC GTGCACAGGGTGTCACGTTGCAAGACCTGCCTGAAACCGAACTGCCCGCTGTTCTGCAGCCGGTCGCGGAGGCCAT GGATGCGATCGCTGCGGCCGATCTTAGCCAGACGAGCGGGTTCGGCCCATTCGGACCGCAAGGAATCGGTCAATAC ACTACATGGCGTGATTTCATATGCGCGATTGCTGATCCCCATGTGTATCACTGGCAAACTGTGATGGACGACACCG TCAGTGCGTCCGTCGCGCAGGCTCTCGATGAGCTGATGCTTTGGGCCGAGGACTGCCCCGAAGTCCGGGCACCTCGT GCACGCGGATTTCGGCTCCAACAATGTCCTGACGGACAATGGCCGCATAACAGCGGTCATTGACTGGAGCGAGGCG ATGTTCGGGGATTCCCAATACGAGGTCGCCAACATCTTCTTCTGGAGGCCGTGGTTGGCTTGTATGGAGCAGCAGA CGCGCTACTTCGAGCGGAGGCATCCGGAGCTTGCAGGATCGCCGCGGCTCCGGGCGTATATGCTCCGCATTGGTCT TGACCAACTCTATCAGAGCTTGGTTGACGGCAATTTCGATGATGCAGCTTGGGCGCAGGGTCGATGCGACGCAATC GTCCGATCCGGAGCCGGGACTGTCGGGCGTACACAAATCGCCCGCAGAAGCGCGGCCGTCTGGACCGATGGCTGTG TAGAAGTACTCGCCGATAGTGGAAACCGACGCCCAGCACTCGTCCGGATCGGGAGATGGGGGAGGCTAACTCTGG TGGCGGTGGCTCGGGCGGAGGTGGGTCGGGTGGCGGCGGATCAATGGCATCCTATCCTTGTCACCAGCACGCGTCT GCATTCGACCAGGCCGCTAGAAGCAGAGGGCACTCCAACCGCAGAACCGCACTGAGACCCCGGAGACAGCAGGAGG CCACAGAAGTGAGGCTGGAGCAGAAGATGCCCACTCTGCTGCGCGTCTACATCGACGGGCCTCACGGAATGGGCAA AACCACAACTACCCAGCTGCTGGTGGCCCTGGGAAGTAGGGACGATATCGTGTACGTCCCTGAGCCAATGACCTAT TGGCAGGTGCTGGGGGCATCAGAAACAATCGCCAACATCTACACAACTCAGCATCGACTGGACCAGGGAGAGATCT CAGCCGGGGATGCAGCTGTGGTCATGACCAGCGCACAGATTACAATGGGCATGCCATATGCAGTGACCGACGCTGT CCTGGCACCACACATCGGAGGAGAAGCAGGAAGCTCCCATGCACCACCTCCAGCTCTGACCCTGATCTTCGATCGG CACCCTATTGCAGCCCTGCTGTGCTACCCAGCTGCAAGATATCTGATGGGCTCTATGACCCCACAGGCCGTGCTGG CTTTTGTCGCACTGATCCCCCCTACACTGCCTGGAACTAATATTGTGCTGGGCGCTCTGCCAGAGGACCGCCATAT CGATCGACTGGCAAAGCGACAGCGACCAGGAGAAAGGCTGGACCTGGCAATGCTGGCCGCTATTAGGCGCGTGTAC GGCCTGCTGGCTAACACTGTCCGGTATCTGCAGGGAGGCGGGTCTTGGAGAGAGGACTGGGGGCAGCTGAGTGGAA CAGCAGTGCCACCCCAGGGAGCCGAACCTCAGAGCAATGCTGGGCCCCGACCTCACATCGGAGATACTCTGTTCAC CCTGTTTCGCGCTCCAGAGCTGCTGGCACCAAACGGCGACCTGTACAACGTGTTCGCATGGGCCCTGGATGTCCTG GCCAAAAGACTGAGGCCCATGCACGTGTTTATTCTGGACTATGATCAGTCCCCTGCAGGATGTCGAGATGCTCTGC TGCAGCTGACTTCCGGCATGGTGCAGACCCATGTCACCACACCCGGGTCTATTCCAACTATTTGTGACCTGGCAAG GACTTTCGCAAGAGAGATGGGAGAGGCTAACAGTGGAGGTGGAGGCAGCGGAGGCGGAGGTTCTGGTGGAGGCGGA AGTATGGTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGG GCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAA GCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAG GCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCG TGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTACAA GGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGGGAGGCCTCC TCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCC ACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACAT CAAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCACTCC ACCGGCGGCATGGACGAGCTGTACAAGTAAACTAGTGGCAGAAGCCTTCAGGTCTGACCCAGCCAATCAAAGCATT ATACCAGACCATGGAATGCATACCAAACATTATTGACACTAATGACACACAAAATTGGTTTTAAGAAAAACCAAGA GAACAATAGGCCAGAATGGCTGGGTCTCGGGAGATATTACTCCCTGAAGTCCATCTCAATTCACCAATTGTAAAGC ATAAGCTATACTATTACATTCTACTTGGAAACCTCCCAAATGAGATCGACCTTGACGATTTAGGTCCATTACATAA TCAAAATTGGAATCAGATAGCACATGAAGAGTCTAACTTAGCTCAACGCTTGGTAAATGTAAGAAATTTTCTAATT ACCCACATCCCTGATCTTAGAAAGGGCCATTGGCAAGAGTATGTCAATGTAATACTGTGGCCGCGAATTCTTCCCT TGATCCCGGATTTTAAAATCAATGACCAATTGCCTCTGCTCAAAAATTGGGACAAGTTAGTTAAAGAATCATGTTC AGTAATCAATGCAGGTACTTCCCAGTGCATTCAGAATCTCAGCTATGGACTGACAGGTCGTGGGAACCTCTTTACA CGATCACGTGAACTCTCTGGTGACCGCAGGGATATTGATCTTAAGACAGTTGTGGCAGCATGGCATGACTCAGACT GGAAAAGAATAAGTGATTTTTGGATTATGATCAAATTCCAGATGAGACAATTAATTGTTAGGCAAACAGATCATAA TGATTCTGATTTAATCACGTATATCGAAAATAGAGAAGGCATAATCATCATAACCCCTGAACTGGTAGCATTATTT AACACTGAGAATCATACACTAACATACATGACCTTTGAAATTGTACTGATGGTTTCAGATATGTACGAAGGTCGTC ACAACATTTTATCACTATGCACAGTTAGCACTTACCTGAATCCTCTGAAGAAAAGAATAACATATTTATTGAGCCT TGTAGATAACTTAGCTTTTCAGATAGGTGATGCTGTATATAACATAATTGCTTTGCTAGAATCCTTTGTATATGCA CAGTTGCAAATGTCAGATCCCATCCCAGAACTCAGAGGACAATTCCATGCATTCGTATGTTCTGAGATTCTTGATG CACTAAGAGGAACTAATAGTTTCACCCAGGATGAATTAAGAACTGTGACAACTAATTTGATATCCCCATTCCAAGA TCTGACCCCAGATCTTACGGCTGAATTGCTCTGTATAATGAGGCTTTGGGGACACCCCATGCTCACTGCCAGTCAA GCTGCAGGAAAGGTACGCGAGTTCTATGTGTGCTGGAAAAGTATTAGACTTTCCCCACCATTATGAAAACACTAGCCT TTTTCCATACTATTCTGATCAATGGATACAGGAGGAAGCATCATGGAGTATGGCCACCCTTAAACTTACCGGGTAA TGCTTCAAAGGGTCTCACGGAACTTATGAATGACAATACTGAAATAAGCTATGAATTCACACTTAAGCATTGGAAG GAAGTCTCTCTTATAAAATTCAAGAAATGTTTTGATGCAGACGCAGGTGAGGAACTCAGTATATTTATGAAAGATA AGGCAATTAGTGCCCCAAAACAAGACTGGATGAGTGTGTTTAGAAGAAGCCTAATCAAACAGCGCCATCAGCATCA TCAGGTCCCCCTACCAAATCCATTCAATCGACGGCTGTTGCTAAACTTTTCTCCGGAGATGACAAATTCGACCCGAAT GTGGAGCTACAGTATGTAACATCAGGTGAGTATCTACATGATGACACGTTTTGTGCATCATATTCACTAAAAGAGA AGGAAATTAAACCTGATGGTCGAATTTTTGCAAAGTTGACTAAGAGAATGAGATCATGTCAAGTTATAGCAGAATC TCTTTAGCGAACCATGCTGGGAAGTTAATGAAAGAGAATGGTGTTGTGATGAATCAGCTATCATTAACAAAATCA CTATTAACAATGAGTCAGATTGGAATAATATCCGAGAAAGCTAGAAAGTCAACTCGAGATAACATAAATCAACCTG GTTTCCAGAATATCCAGAGAAATAAATCACATCACTCCAAGCAAGTCAATCAGCGAGATCCAAGTGATGACTTTGA ATTGGCAGCATCTTTTTTAACTACTGATCTCAAAATATTGTTTACAATGGAGGTACCAGACAATTATCCCATTT GCTCAATCATTAAACAGAATGTATGGTTATTCCTCATCTCTTTGAGTGGATTCACTTACGGCTAATGCGTAGTACAC TTTACGTGGGGGATCCCTTCAACCCACCAGCAGATACCAGTCAATTTGATCTAGATAAAGTAATTAATGGAGATAT CTTCATTGTATCACCCAGAGGTGGAATTGAAGGGCTGTGTCAAAAGGCTTGGACAATGATATCTATCGCTGTGATA ATTCTATCTGCCACAGAGTCTGGCACACGAGTAATGAGTATGGTGCAGGGAGATAATCAAGCAATTGCTGTCACCA CACGAGTACCAAGGAGCCTGCCGACTCTTGAGAAAAAGACTATTGCTTTTAGATCTTGTAATCTATTCTTTGAGAG GTTAAAATGTAATAATTTTGGATTAGGTCACCATTTGAAAGAACAAGAGACTATCATTAGTTCTCACTTCTTTGTT TATAGCAAGAGAATATTCTATCAGGGGAGGATTCTAACGCAAGCCTTAAAAAATGCTAGTAAGCTCTGCTTGACAG CTGATGTCCTAGGAGAATGCACCCAATCATCATGTTCTAATCTTGCAACTACTGTCATGAGGTTAACTGAGAATGG TGTTGAAAAAGATATCTGTTTCTACTTGAATATCTATATGACCATCAAACAGCTCTCCTATGATATCATCTTCCCT CAAGTGTCAATTCCTGGAGATCAGATCACATTAGAATACATAAATAATCCACACCTGGTATCACGATTGGCTCTTT TGCCATCCCAGTTAGGAGGTCTAAACTACCTGTCATGCAGTAGGCTGTTCAATCGAAACATAGGCGACCCGGTGGT TTCCGCAGTTGCAGATCTTAAGAGATTAATTAAATCAGGATGTATGGATTACTGGATCCTTTATAACTTATTAGGG AGAAAACCGGGAAACGGCTCATGGGCTACTTTAGCAGCTGACCCGTACTCAATCAATATAGAGTATCAATACCCTC CAACTACAGCTCTTAAGAGGCACACCCAACAAGCTCTGATGGAACTCAGTACGAATCCAATGTTACGTGGCATATT CTCTGACAATGCACAGGCAGAAGAAAATAACCTTGCTAGGTTTCTCCTGGATAGGGAGGTGATCTTTCCGCGTGTA GCTCACATCATCATTGAGCAAACCAGTGTCGGGAGGAGAAAACAGATTCAAGGATATTTGGATTCAACTAGATCGA TAATGAGGAAATCACTAGAAATTAAGCCCTTATCCAATAGGAAGCTTAATGAAATACTGGATTACAACATCAATTA CCTAGCTTACAATTTGGCATTACTCAAGAATGCTATTGAACCTCCGACTTATTTGAAGGCAATGACACTTGAAACA TGTAGCATCGACATTGCAAGGAACCTCCGGAAGCTCTCCTGGGCCCCACTCTTGGGTGGGAGAAATCTTGAAGGAT TAGAGACGCCAGATCCCATTGAAATTACTGCAGGAGCATTAATTGTTGGATCGGGCTACTGTGAACAGTGTGCTGC AGGAGACAATCGATTCACATGGTTTTTCTTGCCATCTGGTATCGAGATAGGAGGGGATCCCCGTGATAATCCTCCT ATCCGTGTACCGTACATTGGCTCCAGGACTGATGAGAGGAGGGTAGCCTCAATGGCATACATCAGGGGTGCCTCGA GTAGCTTAAAAGCAGTTCTTAGACTGGCGGGAGTGTACATCTGGGCATTCGGAGATACTCTGGAGAATTGGATAGA TGCACTGGATTTGTCTCACACTAGAGTTAACATCACACTTGAACAGCTGCAATCCCTCACCCCACTTCCAACCTCT GCCAATCTAACCCATCGGTTGGATGATGGCACAACTACCCTAAAGTTTACTCCTGCGAGCTCTTACACCTTTTCAA GTTTCACTCATATATCAAATGATGAGCAATACCTGACAATTAATGACAAAACTGCAGATTCAAATATAATCTACCA ACAGTTAATGATCACTGGACTCGGAATCTTAGAAACATGGAATAATCCCCCAATCAATAGAACATTCGAAGAATCT ACCCTACATTTGCACACTGGTGCATCATGTTGTGTCCGACCTGTGGACTCCTGCATTCTCTCAGAAGCATTAACAG TCAAGCCACATATTACAGTACCGTACAGCAATAAATTTGTATTTGATGAGGACCCGCTATCTGAATATGAAACTGC AAAACTGGAATCGTTATTCATTCCAAGCCCAATTAGGCAACATTGATGCTGTAGATATGACAGGTAAATTAACATTA TTGTCCCAATTCACTGCAAGGCAGATTATCAATGCAATCACTGGACTCGATGAGTCTGTCTCTCTTACTAATGATG CCATTGTTGCATCAGACTATGTCTCCAATTGGATTAGTGAATGCATGTATACCAAAATTAGATGAATTATTTATGTA TTGTGGGTGGGAACTACTATTGGAACTATCCTATCAAATGTATTATCTGAGGGTAGTTGGGTGGAGTAATATAGTG GATTATTCTTACATGATCTTGAGAAGAATCCCGGGTGCAGCATTAAACAATCTGGCATCTACATTAAGTCATCCAA AACTTTTCCGACGAGCTATCAACCTAGATATAGTTGCCCCCTTAAATGCTCCTCATTTTGCATCTCTGGACTACAT CAAGATGAGTGTGGATGCAATACTCTGGGGCTGTAAAAGAGTCATCAATGTCTCTCCAATGGAGGGGACTTAGAA TTAGTTGTGACATCTGAAGATAGCCTTATTCTCAGTGACCGATCCATGAATCTCATTGCAAGGAAATTAACTTTAT TATCACTGATTCACCATAATGGTTTGGAACTACCAAAGATTEAAGGGTTCTCTCCTGATGAGAAGTGTTTCGCTTT GACAGAATTTTTGAGGAAAGTGGTGAACTCAGGGTTGAGTTCAATAGAGAACCTATCAATTTTATGTACAATGTG GAGAACCCACGGCTTGCAGCATTCGCCAGCAACAATTACTACCTGACCAGAAAATTATTGAATTCAATACGAGATA CTGAGTCGGGTCAAGTAGCAGTCACCTCATATTATGAATCATTAGAATATATTGATAGTCTTAAGCTAACCCCACA TGTGCCTGGCACCTCATGCATTGAGGATGATAGTCTATGTACAAATGATTACATAATCTGGATCATAGAGTCTAAT GCAAACTTGGAGAAGTATCCAATTCCAAATAGCCCTGAGGATGATTCCAATTTCCATAACTTTAAGTTGAATGCTC CATCGCACCATACCTTACGCCCATTAGGGTTGTCATCAACTGCTTGGTATAAGGGTATAAGCTGCTGCAGGTACCT TGAGCGATTAAAGCTACCACAAGGTGATCATTTATATATTGCAGAAGGTAGTGGTGCCAGTATGACAATCATAGAA TACCTATTCCCAGGAAGAAAGATATATTACAATTCTTTATTTAGTAGTGGTGACAATCCCCCACAAAGAAATTATG CACCAATGCCTACTCAGTTCATTGAGAGTGTCCCATACAAGCTCTGGCAAGCACACACAGATCAATATCCCGAGAT TTTTGAGGACTTCATCCCTCTATGGAACGGAAACGCCGCCATGACTGACATAGGAATGACAGCTTGTGTAGAATTC ATCATCAATCGAGTCGGCCCAAGGACTTGCAGTTTAGTACATGTAGATTTGGAATCAAGTGCAAGCTTAAATCAAC AATGCCTGTCAAAGCCGATAATTAATGCTATCATCACTGCTACAACTGTTTTGTGCCCTCATGGGGTGCTTATTCT GAAATATAGTTGGTTGCCATTTACTAGATTTAGTACTTTGATCACTTTCTTATGGTGCTACTTTGAGAGAATCACT GTTCTTAGGAGCACATATTCTGATCCAGCTAATCATGAGGTTTATTTAATTTGTATCCTTGCCAACAACTTTGCAT TCCAGACTGTCTCGCAGGCAACAGGAATGGCGATGACTTTAACTGATCAAGGGTTTACTTTGATATCACCTGAAAG AATAAATCAGTATTGGGATGGTCACTTGAAGCAAGAACGTATCGTAGCAGAAGCAATTGATAAGGTGGTTCTAGGA GAAAATGCTCTATTTAATTCGAGTGATAATGAATTAATTCTCAAATGTGGAGGGACACCAAATGCACGGAATCTCA TCGATATCGAGCCAGTCGCAACTTTCATAGAATTTGAACAATTGATCTGCACAATGTTGACAACCCACTTGAAGGA AATAATTGATATAACAAGGTCTGGAACCCAGGATTATGAAAGTTTATTACTCACTCCTTACAATTTAGGTCTTCTT GGTAAAATCAGTACGATAGTGAGATTATTAACAGAAAGGATTCTAAATCATACTATCAGGAATTGGTTGATCCTCC CACCTTCGCTCCGGATGATCGTGAAGCAGGACTTGGAATTCGGCATATTCAGGATTACTTCCATCCTCAATTCTGA TCGGTTCCTGAAGCTTTCTCCAAATAGGAAATACTTGATTGCACATTAACTGCAGGCTACATTAGGAAATTGATT GAGGGGGATTGCAATATCGATCTAACCAGACCTATCCAAGCAAATCTGGAAAGCATTAGGTTGTGTAGTCTATT GTCACGATCCAATGGATCAAAGGGAGTCAACAGAGTTTATTGATATAAATATTAATGAAGAAAATAGACCGCGGGAT CGATGGCGAGGAAATCTAAACATATCAAGAATCAGAATTAGTTTAAGAAAAAAGAAGAGGATTAATCTTGGTTTTC CCCTTGGT

[0139] The underlined sequence is the PIV5-AVLP genomic nucleic acid sequence. The italicized and underlined sequences are the entire Pal nucleic acid sequence. The double-underlined sequence is the AOS-modified NTTM-Pal nucleic acid sequence.

[0140] 3. CPI vaccine sequence

[0141] a. pMHD106-CPI-NTTM-OspA BPBPk

[0142] This article provides pMHD106-CPI-NTTM-OspA BPBPk The nucleic acid sequences from 5' to 3' are as follows:

[0143] ACCAGGGGGAAAACGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCTA TGGCCTTCGTGACCGACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCCGACA CAAAAATGTCATCCGTGCTTAAAGCATATGAGCGATTCACACTCACTCAAGAACTGCAAGATCAGAGTGAGGAAGGT ACAATCCCACCTACAACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGATCCCG GCTTCTTCTATTCTGCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACTTACAA TGTTTTCGCTACCATCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATCGAAAGG GTAGAGATCGATGGCTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCTCGTTCAGGTATGAGCCGTGGAGAGAT CAATGCCTATGCTGCACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTTGTTGATTCCGAAGTCG AGGGAACTGCATGGGATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGGATAGTGACT TGCAAGTGCATGACTGCGCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAATATCGTCAGCAAGGCAG GATCAACCCGAGATATCTCCTGCAACCGGAGGCTCGAAGAATAATCCAGAATGTAATCCGAAAGGGAATGGTGGTCA GACATTTCCTCACCTTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTATGCTATGGTAGGG GATGTTGGAAAGTATATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAATATGCATTAGGAACCAGATG GCCCACACTTGCTTTAGCTGCATTTTCAGGAGAGCTAACAAAGCTAAAGTCCCTCATGGCATTATACCAGACCCTTG GTGAGCAGGCCCGATATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTTGCTGCAGCAAACTACCCACTGCTA TATAGCTATGCTATGGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTCTCCAGATCATACATGAA TAAGACATATTTCCAATTGGGAATGGAAACTGCAAGAAAACAACAGGGTGCAGTTGACATGAGGATGGCAGAAGATC TCGGTCTAACTCAAGCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACAGCAAATCGAGGGGCAGAC ACCAGGGGAGGAGTCAACCCGTTCTCATCTATCACTGGGACAACTCAGGTGCCCGCTGCAGCAACAGGTGACACATT CGAGAGTTACATGGCAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCACGATGATGAGATGCCACCAT TGGAAGAGGAGGAAGAGGACGACACATCTGCAGGTCCACGCACTGGACTAACTCTTGAACAAGTGGCCTTGGACATC CAGAACGCAGCAGTTGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGGGTGATCTTGACATCTAGACAAT TCAGATCCCAATCTTAAATCGACACACCTAATTGACCAGTTAGATGGAACTACAGTGGATTCCATAAGGTTCCTGCC TACCATCGGCTTTTAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGATGCCAATAACACAATCCA CAATCTACAATGGATCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATAAGCTCATAGAGACAGGCCTGAATAC TGTGGAGTATTTTACTTCCCAACAAGTCACAGGAACATCCTCTCTTGGAAAGAATACAATACCACCAGGGGTCACAG GACTACTAACCAATGCTGCAGAGGCAAAGATCCAAGAGTCAACCAACCATCAGAAGGGTTCAGTTGGTGGGGGCGCA AAACCAAAGAAACCGCGACCAAAAATTGCCATTGTGCCAGCAGATGACAAAACGGTGCCCGGAAAGCCGATCCCAAA CCCTCTATTAGGTCTGGACTCCACCCCGAGCACCCAAACTGTGCTTGATCTAAGTGGGAAAACATTACCATCAGGAT CCTATAAGGGGGTTAAACTTGCGAAATTTGGAAAAGAAAATCTGATGACACGGTTCATCGAGGAACCCAGAGAGAAT CCTATCGCAACCAATTCCCCCATCGATTTTAAGAGGGGCAGGGATACCGGCGGGTTCCATAGAAGGGAGTACTCAAT CGGATGGGTGGGAGATGAAGTCAAGGTCACTGAGTGGTGCAATCCATCCTGTTCTCCAATCACCGCTGCAGCAAGGC GATTTGAATGCACTTGTCACCAGTGTCCAGTCACTTGCTCTGAATGTGAACGAGATACTTAATACAGTGAGAAATTT GGACTCTCGGATGAATCAACTGGAGACAAAAGTAGATCGCATTCTCTCATCTCAGTCTCTAATCCAGACCATCAAGA ATGACATAGTTGGACTTAAAGCAGGGATGGCTACTTTAGAAGGAATGATTACAACTGTGAAAATCATGGACCCGGGA GTTCCCAGTAATGTTACTGTGGAAGATGTACGCAAGAAACTAAGTAACCATGCTGTTGTTGTGCCAGAATCATTCAA TGATAGTTTCTTGACTCAATCTGAAGATGTAATTTCACTTGATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAGA AGATTGTCAGGAAGGTTCCTCCTCAGAAGGATCTGACTGGATTGAAGATCACACTAGAGCAATTGGCAAAGGATTGC ATCAGCAAACCGAAGATGAGGGAAGAGTATCTCCTCAAAATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCT CAAGAAAGCAATCATCCGCAGTGCAATTTGATCAAGAAACACCCAATTACACTACACTGGTATGACACTGTACTAAC CCTGAGGGTTTTAGAAAAAACGATTAACGATAAATAAGCCCGAACACTACACACCACCCGAGGCAGCCATGCCATCC ATCAGCATCCCCGCAGACCCCACCAATCCACGTCAATCAATAAAAGCGTTCCCAATTGTGATCAACCGTGATGGGGG TGAGAAAGGTCGCTTGGTTAAACAACTACGCACAACCTACTTGAATGACCTAGATACTCATGAGCCACTGGTGACAT TCGTAAATACTTATGGATTCATCTACGAACAGGATCGGGGAAATACCATTGTCGGAGAGGATCAACATGGGAAGAAA AGAGAGGCTGTGACTGCTGCAATGGTTACCCTTGGATGTGGGCCTAATCTACCATCATTAGGGAATGTCCTGGGACA ACTGAGTGAATTCCAGGTCATTGTTAGGAAGACATCCAGCAAAGCGGAAGAGATGGTCTTTGAAATTGTTAAGTATC CGAGAATATTTCGGGGTCATACATTAATCCAGAAAGGACTAGTCTGTGTCTCCGCAGAAAAATTTGTTAAGTCACCA GGGAAAGTACAATCTGGAATGGACTATCTCTTCATTCCGACATTTCTGTCAGTGACTTACTGTCCAGCTGCAATCAA ATTTCAGGTACCTGGCCCCATGTTGAAAATGAGATCAAGATACACTCAGAGCTTACAACTTGAACTAATGATAAGAA TCCTGTGTAAGCCCGATTCGCCACTTATGAAGGTCCATATCCCTGACAAGGAAGGAAGAGGATGTCTTGTATCAGTA TGGTTGCATGTATGCAATATCTTCAAATCAGGAAACAAGAATGGCAGTGAGTGGCAGGAATACTGGATGAGAAAGTG TGCTAACATGCAACTTGAAGTGTCGATTGCAGATATGTGGGGACCAACTATCATAATTCATGCCAGAGGTCACATTC CCAAAAGTGCTAAGTTGTTTTTTGGAAAGGGTGGATGGAGCTGCCATCCACTTCACGAAGTTGTTCCAAGTGTCACT AAAACACTATGGTCCGTGGGCTGTGAGATTACAAAGGCGAAGGCAATAATACAAGAGAGTAGCATCTCTCTTCTCGT GGAGACTACTGACATCATAAGTCCAAAAGTCAAAATTTCATCTAAGCATCGCCGCTTTGGGAAATCAAATTGGGGTC TGTTCAAGAAAACCAAATCACTGCCCAACCTGACGGAGCTGGAATGACTGACCCCCAATCGAGACTACACCACCTCA AACTATAGGTGGGTGGTACCTCAGTGATTAATCTCGTAAGCACTGATCGTAGGCTACAACACACTAATATTATCCAG ATTAGAGAGCTTAATTAGCTCTGTATTAATAATAACACTACTATTCCAATAACTGGAATCACCAGCTTGATTTATCT CCAAAATGATTCAAAGAAAACAAATCATATTAAGACTATCCTAAGCACGAACCCATATCGTCCTTCAAATCATGGGT ACTATAATTCAATTTCTGGTGGTCTCCTGTCTATTGGCAGGAGCAGGCAGTCTTGATCCAGCAGCCCTCATGCAAAT CGGTGTCATTCCAACAAATGTCCGGCAACTTATGTATTATACTGAGGCCTCATCAGCATTCATTGTTGTGAAGTTAA TGCCTACAATTGACTCGCCGATTAGTGGATGTAATATAACATCAATTTCAAGCTATAATGCAACAGTGACAAAACTC CTACAGCCGATCGGTGAGAATTTGGAGACGATTAGGAACCAGTTGATTCCAACTCGGAGGAGACGCCGGTTTGCAGG GGTGGTGATTGGATTAGCTGCATTAGGAGTAGCTACTGCCGCACAGGTCACTGCCGCAGTAGCACTAGTAAAGGCAA ATGAAAATGCTGCGGCTATACTCAATCTCAAAAATGCAATCCAAAAAACAAATGCAGCAGTTGCAGATGTGGTCCAG GCCACACAATCACTAGGAACAGCAGTTCAAGCAGTTCAAGATCACATAAACAGTGTGGTAAGTCCAGCAATTACAGC AGCCAATTGTAAAGCCCAAGATGCTATCATTGGCTCAATCCTCAATCTCTATTTGACCGAGTTGACAACTATCTTCC ACAATCAAATTACAAACCCTGCATTGAGTCCTATTACAATTCAAGCTTTAAGGATCCTACTAGGGAGTACCTTGCCG ACTGTGGTCGAAAAATCTTTCAATACCCAGATAAGTGCGGCTGAGCTTCTCTCATCAGGGTTATTGACAGGCCAGAT TGTGGGATTAGATTTGACCTATATGCAGATGGTCATAAAAATTGAGCTGCCAACTTTAACTGTACAACCTGCAACCC AGATCATAGATCTGGCCACCATTTCTGCATTCATTAACAATCAAGAAGTCATGGCCCAATTACCAACACGTGTTATT GTGACTGGCAGCTTGATCCAAGCCTATCCCGCATCGCAATGCACTATTACACCCAACACTGTGTACTGTAGGTATAA TGATGCCCAAGTACTCTCAGATGATACGATGGCTTGCCTCCAAGGTAACTTGACAAGATGCACCTTCTCTCCAGTGG TTGGGAGCTTTCTCACTCGATTCGTGCTGTTCGATGGAATAGTTTATGCAAATTGCAGGTCGATGTTGTGCAAGTGC ATGCAGCCTGCTGCTGTGATCCTACAGCCGAGTTCATCCCCTGTAACTGTCATTGACATGTACAAATGTGTGAGTCT GCAGCTTGACAATCTCAGATTCACCATCACTCAATTGGCCAATGTAACCTACAATAGCACCATCAAGCTTGAAACAT CCCAGATCTTGCCTATTGATCCGTTGGATATATCCCAGAATCTAGCTGCGGTGAATAAGAGTCTAAGTGATGCACTA CAACACTTAGCACAAAGTGACACATACCTTTCTGCAATCACATCAGCTACGACTACAAGTGTATTATCCATAATAGC AATCTGTCTTGGATCGTTAGGTTTAATATTAATAATCTTGCTCAGTGTAGTTGTGTGGAAGTTATTGACCATTGTCG CTGCTAATCGAAATAGAATGGAGAATTTTGTTTATCATAATTCAGCATTCCACCACTCACGATCTGATCTCAGTGAG AAAAATCAACCTGCAACTCTTGGAACAAGATAAGACAGTCATCCATTAGTAATTTTTAAGAAAAAAACGATAGGACC GAAACTAGTATTGAAAGAACCGTCTCGGTCAATCTAGGTAATCGAGCTGCTACCGTCTCGGAAAGCTCAAATCATGC TGCCTGATCCGGAAGATCCGGAAAGCAAAAAAGCTACAAGGAGAACAGGAAACCTAATTATCTGCTTCCTATTCATC TTCTTTCCGTTTGTAAACTTCATTGTTCCAACTCTAAGACACTTGCTGTCCTAACACCTGCTATAGGCTATCCACTG CATCATCTCTTTTTAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGGTGCCAACA

[0144]

[0145] GATCCTGGCCCTGATCGCCTGCAAGCAGAACGTGAGCTCCCTGGACGAGAAGAATAGCGTGTCCGTGG ATCTGCCCGGCGAGATGAAGGTGCTGGTGAGCAAGGAGAAGAACAAGGACGGCAAGTATGATCTGATCGCCACAGT GGACAAGCTGGAGCTGAAGGGCACCAGCGATAAGAACAATGGATCCGGCGTGCTGGAGGGAGTGAAGGCAGACAAG TCTAAGGTGAAGCTGACCATCAGCGACGATCTGGGCCAGACCACACTGGAGGTGTTCAAGGAGGACGGCAAGACCC TGGTGTCCAAGAAGGTGACATCTAAGGATAAGTCTAGCACCGAGGAGAAGTTTAACGAGAAGGGCGAGGTGTCCGA GAAGATCATCACACGGGCCGACGGCACCAGACTGGAGTACACAGGCATCAAGTCCGATGGCTCTGGCAAGGCCAAG GAGGTGCTGAAGGGCTTCACACTGGAGGGCAAGGTGGCCAATGATAAGGTGACCCTGGAGGTGAAGGAGGGCACCG TGACACTGTCCAAGAACATCTCTAAGAGCGGCGAGGTGTCTGTGGAGCTGAATGACACCGATTCCTCTGCCGCCAC AAAGAAGACCGCCGCCTGGAACAGCAAGACCTCCACACTGACCATCTCTGTGAATAGCAAGAAGACCACACAGCTG GTGTTTACAAAGCAGGACACAATCACCGTGCAGAAGTATGATAGCGCCGGAACCAACCTGGAGGGAACAGCAGTGG AGATCAAGACCCTGGACGAGCTGAAGAATGCCCTGAAGTGAGCTAGCCTCCTGCCATACTTCCTACTCACATCATA TCTATTTTAAAGAAAAAATAGGCCCGAACACTAATCGTGCCGGCAGTGCCCTGCACACACAACACTACACATACAA TACACTACAATGGTTGCAGAAGATGCCCCTGTTAGGGGCACTTGCCGAGTATTATTTCGAACAACAACTTTAATTT TTCTATGCACACTATTAGCATTAAGCATCTCTATCCTTTATGAGAGTTTAATAACCCAAAAGCAAATCATGAGCCA AGCAGGCTCAACTGGATCTAATTCTAGATTAGGAAGTATCACTGATCTTCTTAATAATATTCTCTCTGTCGCAAAT CAGATTATATATAACTCTGCAGTCGCTCTACCTCTACAATTGGACACTCTTGAATCAACACTCCTTACAGCCATTA AGTCTCTTCAAACAAGTGACAAGCTAGAACAGAACTGCTCGTGGGGTGCTGCACTGATTAATGATAATAGATACAT TAATGGCATCAATCAGTTCTATTTCTCAATTGCTGAGGGTCGCAAGCTGACACTTGGCCCACTTCTTAATATACCT AGTTTCATTCCAACTGCCACGACACCAGAGGGCTGCACCAGGATCCCATCATTCTCGCTCACCAAGACACACTGGT GTTATACACACAATGTTATCCTGAATGGATGCCAGGATCATGTATCCTCAAATCAATTTGTTTCCATGGGAATCAT TGAACCCACTTCTGCCGGGTTTCCATCCTTTCGAACCCTAAAGACTCTATATCTCAGCGATGGGGTCAATCGTAAG AGCTGCTCTATCAGTACAGTTCCGGGGGGTTGTATGATGTACTGTTTCGTCTCTACTCAACCAGAGAGGGATGACT ACTTTTCTACCGCTCCTCCAGAACAACGAATTATTATAATGTACTATAATGATACAATCGTGGAGCGCATAATTAA TCCACCCGGGGTACTAGATGTATGGGCAACATTGAACCCAGGAACAGGAAGCGGGGTATATTATTTAGGTTGGGTA CTCTTTCCAATATATGGCGGCGTGATTAAAGATACGAGTTTATGGAATAATCAAGCAAATAAATACTTTATCCCCC AGATGGTTGCTGCTCTCTGCTCACAAAACCAGGCAACTCAAGTCCAAAATGCTAAGTCATCATACTATAGCAGCTG GTTTGGCAATCGAATGATTCAGTCTGGGATCCTGGCATGTCCTCTTCAACAGGATCTAACCAATGAGTGTTTAGTT CTGCCCTTTTCTAATGATCAGGTGCTTATGGGTGCTGAAGGGAGATTATACATGTATGGTGACTCGGTGTATTACT ACCAAAGAAGCAATAGTTGGTGGCCTATGACCATGCTGTATAAGGTAACCATAACATTCACTAATGGTCAGCCATC TGCTATATCAGCTCAGAATGTGCCCACACAGCAGGTCCCTAGACCTGGGACAGGAAGCTGCTCTGCAACAAATAGA TGTCCCGGTTTTTGCTTGAAAGGAGTGTATGCTGATGCCTGGTTACTGACCAACCCTTCGTCTACCAGTACATTTG GATCAGAAGCAACCTTCACTGGTTCTTATCTCAACGCAGCAACTCAGCGTATCAATCCGACGATGTATATCGCGAA CAACACACAGATCATAAGCTCACAGCAATTTGGATCAAGCGGTCAAGAAGCAGCATATGGCCACACAACTTGTTTT AGGGACACAGGCTCTGTTATGGTATACTGTATCTATATTATTGAATTGTCCTCATCTCTCTTAGGACAATTTCAGA TTGTCCCATTTATCCGTCAGGTGACACTATCCTAAAGGCAGAAGCCTCCAGGTCTGACCCAGCCAATCAAAGCATT ATACCAGACCATGGAATGCATACCAAACATTATTGACACTAATGACACACAAAATTGGTTTTAAGAAAAACCAAGA GAACAATAGGCCAGAATGGCTGGGTCTCGGGAGATATTACTCCCTGAAGTCCATCTCAATTCACCAATTGTAAAGC ATAAGCTATACTATTACATTCTACTTGGAAACCTCCCAAATGAGATCGACATTGACGATTTAGGTCCATTACATAA TCAAAATTGGAATCAAATAGCACATGAAGAGTCTAACTTAGCCCAACGCTTGGTAAATGTAAGAAATTTTCTAATT ACCCACATCCCTGATCTTAGAAAGGGCCATTGGCAAGAGTATGTCAATGTAATACTGTGGCCGCGAATTCTTCCCT TGATCCCGGATTTTAAAATCAATGACCAATTGCCTCTACTCAAAAATTGGGACAAGTTAGTTAAAGAATCATGTTC AGTAATCAATGCGGGTACTTCCCAGTGCATTCAGAATCTCAGCTATGGACTGACAGGTCGTGGGAACCTCTTTACA CGATCACGTGAACTCTCTGGTGACCGCAGGGATATTGATCTTAAGACGGTTGTGGCAGCATGGCATGACTCAGACT GGAAAAGAATAAGTGATTTTTGGATTATGATCAAATTCCAGATGAGACAATTAATTGTTAGGCAAACAGATCATAA TGATCCTGATTTAATCACGTATATCGAAAATAGAGAAGGCATAATCATCATAACCCCTGAACTGGTAGCATTATTT AACACTGAGAATCATACACTAACATACATGACCTTTGAAATTGTACTGATGGTTTCAGATATGTACGAAGGTCGTC ACAACATTTTATCACTATGCACAGTTAGCACTTACCTGAATCCTCTGAAGAAAAGAATAACATATTTATTGAGCCT TGTAGATAACTTAGCTTTTCAGATAGGTGATGCTGTATATAACATAATTGCTTTGCTAGAATCCTTTGTATATGCA CAGTTGCAAATGTCAGATCCCATCCCAGAACTCAGAGGACAATTCCATGCATTCGTATGTTCTGAGATTCTTGATG CACTAAGGGGAACTAATAGTTTCACCCAGGATGAATCAAGAACTGTGACAACCAATTTGATATCCCCATTCCAAGA TCTGACCCCAGATCTTACGGCTGAATTGCTCTGTATAATGAGGCTTTGGGGACACCCCATGCTCACCGCCAGTCAA GCTGCGGGAAAGGTACGCGAGTCCATGTGTGCTGGAAAAGTATTAGACTTTCCCACCATTATGAAAACACTAGCCT TTTTCCATACTATTCTGATCAATGGATACAGGAGGAAGCATCATGGAGTATGGCCACCCTTAAACTTACCGGGTAA TGCTTCAAAGGGTCTCACGGAACTTATGAATGACAATACTGAGATAAGCTATGAATTCACACTTAAGCATTGGAAG GAAATCTCTCTTATAAAATTCAAGAAATGTTTTGATGCAGACGCAGGTGAGGAACTCAGTATATTTATGAAAGATA AAGCAATTAGTGCCCCAAAACAAGATTGGATGAGTGTGTTTAGAAGAAGCCTAATCAAACAGCGCCATCAGCATCA TCAGGTCCCCCTACCAAATCCATTCAATCGACGGCTATTGCTAAACTTTCTCGGAGATGACAAATTCGACCCGAAT GTGGAGCTACAGTATGTAACATCAGGTGAGTATCTACATGATGACACGTTTTGTGCATCATATTCACTAAAAGAGA AGGAAATTAAACCTGATGGTCGAATTTTTGCAAAGTTGACTAAGAGAATGAGATCATGTCAAGTTATAGCAGAATC TCTTTAGCGAACCATGCTGGGAAGTTAATGAAAGAGAATGGTGTTGTGATGAATCAGCTATCATTAACAAAATCA CTATTAACAATGAGTCAGATTGGAATAATATCCGAGAAAGCTAGAAAATCGACTCGAGATAACATAAATCAACCTG GTTTCCAGAATATCCAGAGAAATAAATCACATCACTCCAAGCAAGTCAATCAGCGAGATCCAAGTGATGACTTTGA ATTGGCAGCATCTTTTTTAACTACTGATCTCAAAATATTGTTTACAATGGAGGTACCAGACAATTATCCCATTT GCTCAATCATTAAACAGAATGTATGGTTATTCCTCATCTCTTTGAGTGGATTCACTTACGGCTAATGCGTAGTACAC TTTACGTGGGGGATCCCTTCAACCCACCAGCAGATACCAGTCAATTTGATCTAGATAAAGTAATTAATGGAGATAT CTTCATTGTATCACCCAGAGGTGGAATTGAAGGGCTATGTCAAAAGGCTTGGACAATGATATCTATCTCTGTGATA ATTCTATTCTGCCACAGAGTCTGGCACACGAGTAATGAGTATGGTGCAGGGAGATAATCAAGCAATTGCTGTCACCA CACGAGTACCAAGGAGCCTGCCGACTCTTGAGAAAAAGACTATTGCTTTAGATCTTGTAATCTATTCTTTGAGAG GTTAAAATGTAATAATTTTGGATTAGGTCACCATTTGAAAGAACAAGAGACTATCATTAGTTCTCACTTCTTTGTT TATAGCAAGAGAATATTCTATCAGGGGAGGATTCTAACGCAAGCCTTAAAAAATGCTAGTAAGCTCTGCTTGACAG CTGATGTCCTAGGAGAATGTACCCAATCATCATGTTCTAATCTTGCAACTACTGTCATGAGGTTAACTGAGAATGG TGTTGAAAAAGATATCTGTTTCTACTTGAATATCTATATGACCATCAAACAGCTCTCCTATGATATCATCTTCCCT CAAGTGTCAATTCCTGGAGATCAGATCACATTAGAATACATAAATAATCCACACCTGGTATCACGATTGGCTCTTC TGCCATCCCAGCTAGGAGGTCTAAACTACCTGTCATGCAGTAGGCTGTTCAATCGAAACATAGGCGACCCGGTGGT TTCCGCAGTTGCAGATCTTAAGAGATTAATTAAATCAGGATGTATGGATTACTGGATCCTTTATAACTTATTAGGG AGAAAACCGGGAAACGGCTCATGGGCTACTTTAGCAGCTGACCCGTACTCAATCAATATAGAGTATCAATACCCCC CAACTACAGCTCTTAAGAGGCACACCCAACAAGCTCTGATGGAACTCAGTACGAATCCAATGTTACGTGGCATATT CTCTGACAATGCACAGGCAGAAGAAAATAATCTTGCTAGATTTCTCCTGGATAGGGAGGTGATCTTTCCGCGTGTA GCTCACATCATCATTGAGCAAACCAGTGTCGGGAGGAGAAAACAGATTCAAGGATATTTGGATTCAACTAGATCGA TAATGAGTAAATCACTAGAAATTAAGCCCTTGTCCAATAGGAAGCTTAATGAAATACTGGATTACAACATCAATTA CCTAGCTTACAATTTGGCATTACTCAAGAATGCTATTGAACCTCCGACTTATTTGAAAGCAATGACTCTTGAAACA TGTAGCATCGACATTGCAAGGAGCCTCCGGAAGCTCTCCTGGGCCCCACTCTTGGGTGGGAGAAATCTTGAAGGAT TAGAGACGCCAGATCCCATTGAAATTACTGCAGGAGCATTAATTGTTGGATCGGGCTACTGTGAACAGTGTGCTGC AGGAGACAATCGATTCACATGGTTTTTCTTGCCATCTGGTATCGAGATAGGAGGGGATCCCCGTGATAATCCTCCT ATCCGTGTACCGTACATTGGCTCCAGGACTGATGAGAGGAGGGTAGCCTCAATGGCATACATCAGGGGTGCCTCGA GTAGCCTAAAAGCAGTTCTTAGACTGGCGGGAGTGTACATCTGGGCATTCGGAGATACTCTGGAGAATTGGATAGA TGCACTGGATTTGTCTCACACTAGAGTTAACATCACACTTGAACAGCTGCAATCCCTCACCCCACTTCCAACCTCT GCCAATCTAACCCATCGGTTGGATGATGGCACAACTACCCTAAAGTTTACTCCTGCGAGCTCTTACACCTTTTCAA GTTTCACTCATATATCAAATGATGAGCAATACCTGACAATTAATGACAAAACTGCAGATTCAAATATAATCTACCA ACAGTTAATGATCACTGGACTCGGAATTTTAGAAACATGGAATAATCCCCCAATCAATAGAACATTCGAAGAATCT ACCCTACATTTGCACACTGGTGCATCATGTTGTGTCCGACCTGTGGACTCCTGCATCATCTCAGAAGCATTAACAG TCAAGCCACATATTACAGTACCGTACAGCAATAAATTTGTATTTGATGAAGACCCGCTATCTGAATATGAGACTGC AAAACTGGAATCGTTATCATTTCAAGCCCAATTAGGCAACATTGATGCTGTAGATATGACAGGTAAATTAACATTA TTGTCCCAATTCACTGCAAGGCAGATTATTAATGCAATCACTGGACTCGATGAGTCTGTCTCTCTTACTAATGATG CCATTGTTGCATCAGACTATGTCTCCAATTGGATTAGTGAATGCATGTATACCAAAATTAGATGAATTATTTATGTA TTGTGGGTGGGAACTACTATTGGAACTATCCTATCAAATGTATTATCTGAGGGTAGTTGGGTGGAGTAATATAGTG GATTATTCTTACATGATCTTGAGAAGAATCCCGGGTGCAGCATTAAACAATCTGGCATCTACATTAAGTCATCCAA AACTTTTCCGACGAGCTATCAACCTAGATATAGTTGCCCCCTTAAATGCTCCTCATTTTGCATCTCTGGACTACAT CAAGATGAGTGTGGATGCAATACTCTGGGGCTGTAAAAGAGTCATCAATGTCTCTCCAATGGAGGGGACTTAGAA TTAGTTGTGACATCTGAAGATAGCCTTATTCTCAGTGACCGATCCATGAATCTCATTGCAAGGAAATTAACTTTAT TATCACTGATTCACCATAATGGTTTGGAACTACCAAAGATTEAAGGGTTCTCTCCTGATGAGAAGTGTTTCGCTTT GACAGAATTTTTGAGGAAAGTGGTGAACTCAGGGTTGAGTTCAATAGAGAACCTATCAATTTTATGTACAATGTG GAGAACCCACGGCTTGCAGCATTCGCCAGCAACAATTACTACCTGACCAGAAAATTATTGAATTCAATACGAGATA CTGAGTCGGGTCAAGTAGCAGTCACCTCATATTATGAATCATTAGAATATATTGATAGTCTTAAGCTAACCCCACA TGTGCCTGGTACCTCATGCATTGAGGATGATAGTCTATGTACAAATGATTACATAATCTGGATCATAGAGTCTAAT GCAAACTTGGAGAGTATCCAATTCCAAATAGCCCTGAGGATGATTCCAATTTCCATAACTTTAAGTTGAATGCTC CATCGCACCATACCTTACGCCCATTAGGGTTGTCATCAACTGCTTGGTATAAGGGTATAAGCTGTTGCAGGTACCT TGAGCGATTAAAGCTACCACAAGGTGATCATTTATATATTGCAGAAGGTAGTGGTGCCAGTATGACAATCATAGAA TACCTATTCCCAGGAAGAAAGATATATTACAATTCTTTATTTAGTAGTGGTGACAATCCCCCACAAAGAAATTATG CACCAATGCCTACTCAGTTCATTGAGAGTGTCCCATACAAGCTCTGGCAAGCACACACAGATCAATATCCCGAGAT TTTTGAGGACTTCATCCCTCTATGGAACGGAAATGCCGCCATGACTGACATAGGAATGACAGCTTGTGTAGAATTC ATCATCAATCGAGTCGGCCCAAGGACTTGCAGTTTAGTACATGTAGATTTGGAATCAAGTGCAAGCTTAAATCAAC AATGCCTGTCAAAGCCGATAATTAATGCTATCATCACTGCTACAACTGTTTTGTGCCCTCATGGGGTGCTTATTCT GAAATATAGTTGGTTGCCATTTACTAGATTTAGTACTTTGATCACTTTCTTATGGTGCTACTTTGAGAGAATCACT GTTCTTAGGAGCACATATTCTGGTCCAGCTAATCATGAGGTTTATTTAATTTGTATCCTTGCCAACAACTTTGCAT TCCAGACTGTCTCGCAGGCAACAGGAATGGCGATGACTTTAACCGATCAAGGGTTTACTTTGATATCACCTGAAAG AATAAATCAGTATTGGGATGGTCACTTGAAGCAAGAACGTATCGTAGCAGAAGCAATTGATAAGGTGGTTCTAGGA GAAGATGCTCTATTCAATTCGAGTGATAATGAATTAATTCTCAAATGTGGAGGGACACCAAATGCACGGAATCTTA TCGATATCGAGCCAGTCGCAACTTTCATAGAATTTGAACAACTGATCTGCACAATGTTGACAACCCACTTGAAGGA AATAATTGATATAACAAGGTCTGGAACCCAGGATTATGAAAGTTTATTACTCACTCCTTACAATTTAGGTCTTCTT GGTAAAATCAGTACGATAGTGAGATTATTAACAGAAAGGATTCTAAATCATACTATCAGGAATTGGTTGATCCTCC CACCTTCGCTCCGGATGATCGTGAAGCAGGACTTGGAATTCGGCATATTCAGGATTACTTCCATCCTCAATTCTGA TCGGTTCCTGAAGCTTTCTCCAAATAGGAAATACTTGATTACACAATTAACTGCAGGCTACATTAGGAAATTGATT GAGGGGGATTGTAATATCGATCTAACCAGACCTATCCAAAAACAAATCTGGAAAGCATTAGGTTGTGTAGTCTATT GTCACGATCCAGTAGATCAAAGGGAATCAACAGAGTTTATTGATATAAATATTAATGAAGAAATAGACCGCGGGAT CGATGGCGAGGAAATCTAAATATATCAAGAATCAGAATTAGTTTAAGAAAAAAGAAGAGGATTAATCTTGGTTTTC CCCTTGGT

[0146] The underlined sequence is the PIV5-CPI genomic nucleic acid sequence. The italicized and underlined sequences are the entire OspA nucleic acid sequence. The double-underlined sequence is AOS-modified NTTM-OspA. BPBPk Nucleic acid sequence.

[0147] b. pMHD175-CPI-MOMPΔsp-CD5sp

[0148] The pMHD175-CPI-MOMPΔsp-CD5sp nucleic acid sequence provided in this article, from 5' to 3', is as follows:

[0149] ACCAGGGGGAAAACGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCTA TGGCCTTCGTGACCGACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCGACA CAAAAATGTCATCCGTGCTTAAAGCATATGAGCGATTCACACTCACTCAAGAACTGCAAGATCAGAGTGAGGAAGGT ACAATCCCACCTACAACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGATCCCG GCTTCTTCTATTCTGCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACTTACAA TGTTTTCGCTACCATCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATCGAAAGG GTAGAGATCGATGGCTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCTCGTTCAGGTATGAGCCGTGGAGAGAT CAATGCCTATGCTGCACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTTGTTGATTCCGAAGTCG AGGGAACTGCATGGGATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGGATAGTGACT TGCAAGTGCATGACTGCGCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAATATCGTCAGCAAGGCAG GATCAACCCGAGATATCTCCTGCAACCGGAGGCTCGAAGAATAATCCAGAATGTAATCCGAAAGGGAATGGTGGTCA GACATTTCCTCACCTTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTATGCTATGGTAGGG GATGTTGGAAAGTATATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAATATGCATTAGGAACCAGATG GCCCACACTTGCTTTAGCTGCATTTTCAGGAGAGCTAACAAAGCTAAAGTCCCTCATGGCATTATACCAGACCCTTG GTGAGCAGGCCCGATATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTTGCTGCAGCAAACTACCCACTGCTA TATAGCTATGCTATGGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTCTCCAGATCATACATGAA TAAGACATATTTCCAATTGGGAATGGAAACTGCAAGAAAACAACAGGGTGCAGTTGACATGAGGATGGCAGAAGATC TCGGTCTAACTCAAGCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACAGCAAATCGAGGGGCAGAC ACCAGGGGAGGAGTCAACCCGTTCTCATCTATCACTGGGACAACTCAGGTGCCCGCTGCAGCAACAGGTGACACATT CGAGAGTTACATGGCAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCACGATGATGAGATGCCACCAT TGGAAGAGGAGGAAGAGGACGACACATCTGCAGGTCCACGCACTGGACTAACTCTTGAACAAGTGGCCTTGGACATC CAGAACGCAGCAGTTGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGGGTGATCTTGACATCTAGACAAT TCAGATCCCAATCTTAAATCGACACACCTAATTGACCAGTTAGATGGAACTACAGTGGATTCCATAAGGTTCCTGCC TACCATCGGCTTTTAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGATGCCAATAACACAATCCA CAATCTACAATGGATCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATAAGCTCATAGAGACAGGCCTGAATAC TGTGGAGTATTTTACTTCCCAACAAGTCACAGGAACATCCTCTCTTGGAAAGAATACAATACCACCAGGGGTCACAG GACTACTAACCAATGCTGCAGAGGCAAAGATCCAAGAGTCAACCAACCATCAGAAGGGTTCAGTTGGTGGGGGCGCA AAACCAAAGAAACCGCGACCAAAAATTGCCATTGTGCCAGCAGATGACAAAACGGTGCCCGGAAAGCCGATCCCAAA CCCTCTATTAGGTCTGGACTCCACCCCGAGCACCCAAACTGTGCTTGATCTAAGTGGGAAAACATTACCATCAGGAT CCTATAAGGGGGTTAAACTTGCGAAATTTGGAAAAGAAAATCTGATGACACGGTTCATCGAGGAACCCAGAGAGAAT CCTATCGCAACCAATTCCCCCATCGATTTTAAGAGGGGCAGGGATACCGGCGGGTTCCATAGAAGGGAGTACTCAAT CGGATGGGTGGGAGATGAAGTCAAGGTCACTGAGTGGTGCAATCCATCCTGTTCTCCAATCACCGCTGCAGCAAGGC GATTTGAATGCACTTGTCACCAGTGTCCAGTCACTTGCTCTGAATGTGAACGAGATACTTAATACAGTGAGAAATTT GGACTCTCGGATGAATCAACTGGAGACAAAAGTAGATCGCATTCTCTCATCTCAGTCTCTAATCCAGACCATCAAGA ATGACATAGTTGGACTTAAAGCAGGGATGGCTACTTTAGAAGGAATGATTACAACTGTGAAAATCATGGACCCGGGA GTTCCCAGTAATGTTACTGTGGAAGATGTACGCAAGAAACTAAGTAACCATGCTGTTGTTGTGCCAGAATCATTCAA TGATAGTTTCTTGACTCAATCTGAAGATGTAATTTCACTTGATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAGA AGATTGTCAGGAAGGTTCCTCCTCAGAAGGATCTGACTGGATTGAAGATCACACTAGAGCAATTGGCAAAGGATTGC ATCAGCAAACCGAAGATGAGGGAAGAGTATCTCCTCAAAATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCT CAAGAAAGCAATCATCCGCAGTGCAATTTGATCAAGAAACACCCAATTACACTACACTGGTATGACACTGTACTAAC CCTGAGGGTTTTAGAAAAAACGATTAACGATAAATAAGCCCGAACACTACACACCACCCGAGGCAGCCATGCCATCC ATCAGCATCCCCGCAGACCCCACCAATCCACGTCAATCAATAAAAGCGTTCCCAATTGTGATCAACCGTGATGGGGG TGAGAAAGGTCGCTTGGTTAAACAACTACGCACAACCTACTTGAATGACCTAGATACTCATGAGCCACTGGTGACAT TCGTAAATACTTATGGATTCATCTACGAACAGGATCGGGGAAATACCATTGTCGGAGAGGATCAACATGGGAAGAAA AGAGAGGCTGTGACTGCTGCAATGGTTACCCTTGGATGTGGGCCTAATCTACCATCATTAGGGAATGTCCTGGGACA ACTGAGTGAATTCCAGGTCATTGTTAGGAAGACATCCAGCAAAGCGGAAGAGATGGTCTTTGAAATTGTTAAGTATC CGAGAATATTTCGGGGTCATACATTAATCCAGAAAGGACTAGTCTGTGTCTCCGCAGAAAAATTTGTTAAGTCACCA GGGAAAGTACAATCTGGAATGGACTATCTCTTCATTCCGACATTTCTGTCAGTGACTTACTGTCCAGCTGCAATCAA ATTCAGGTACCTGGCCCCATGTTGAAAATGAGATCAAGATACACTCAGAGCTTACAACTTGAACTAATGATAAGAA TCCTGTGTAAGCCCGATTCGCCACTTATGAAGGTCCATATCCCTGACAAGGAAGGAAGAGGATGTCTTGTATCAGTA TGGTTGCATGTATGCAATATCTTCAAATCAGGAAACAAGAATGGCAGTGAGTGGCAGGAATACTGGATGAGAAAGTG TGCTAACATGCAACTTGAAGTGTCGATTGCAGATATGTGGGGACCAACTATCATAATTCATGCCAGAGGTCACATTC CCAAAAGTGCTAAGTTGTTTTTTGGAAAGGGTGGATGGAGCTGCCATCCACTTCACGAAGTTGTTCCAAGTGTCACT AAAACACTATGGTCCGTGGGCTGTGAGATTACAAAGGCGAAGGCAATAATACAAGAGAGTAGCATCTCTCTTCTCGT GGAGACTACTGACATCATAAGTCCAAAAGTCAAAATTTCATCTAAGCATCGCCGCTTTGGGAAATCAAATTGGGGTC TGTTCAAGAAAACCAAATCACTGCCCAACCTGACGGAGCTGGAATGACTGACCCCCAATCGAGACTACACCACCTCA AACTATAGGTGGGTGGTACCTCAGTGATTAATCTCGTAAGCACTGATCGTAGGCTACAACACACTAATATTATCCAG ATTAGAGAGCTTAATTAGCTCTGTATTAATAATAACACTACTATTCCAATAACTGGAATCACCAGCTTGATTTATCT CCAAAATGATTCAAAGAAAACAAATCATATTAAGACTATCCTAAGCACGAACCCATATCGTCCTTCAAATCATGGGT ACTATAATTCAATTTCTGGTGGTCTCCTGTCTATTGGCAGGAGCAGGCAGTCTTGATCCAGCAGCCCTCATGCAAAT CGGTGTCATTCCAACAAATGTCCGGCAACTTATGTATTATACTGAGGCCTCATCAGCATTCATTGTTGTGAAGTTAA TGCCTACAATTGACTCGCCGATTAGTGGATGTAATATAACATCAATTTCAAGCTATAATGCAACAGTGACAAAACTC CTACAGCCGATCGGTGAGAATTTGGAGACGATTAGGAACCAGTTGATTCCAACTCGGAGGAGACGCCGGTTTGCAGG GGTGGTGATTGGATTAGCTGCATTAGGAGTAGCTACTGCCGCACAGGTCACTGCCGCAGTAGCACTAGTAAAGGCAA ATGAAAATGCTGCGGCTATACTCAATCTCAAAAATGCAATCCAAAAAACAAATGCAGCAGTTGCAGATGTGGTCCAG GCCACACAATCACTAGGAACAGCAGTTCAAGCAGTTCAAGATCACATAAACAGTGTGGTAAGTCCAGCAATTACAGC AGCCAATTGTAAAGCCCAAGATGCTATCATTGGCTCAATCCTCAATCTCTATTTGACCGAGTTGACAACTATCTTCC ACAATCAAATTACAAACCCTGCATTGAGTCCTATTACAATTCAAGCTTTAAGGATCCTACTAGGGAGTACCTTGCCG ACTGTGGTCGAAAAATCTTTCAATACCCAGATAAGTGCGGCTGAGCTTCTCTCATCAGGGTTATTGACAGGCCAGAT TGTGGGATTAGATTTGACCTATATGCAGATGGTCATAAAAATTGAGCTGCCAACTTTAACTGTACAACCTGCAACCC AGATCATAGATCTGGCCACCATTTCTGCATTCATTAACAATCAAGAAGTCATGGCCCAATTACCAACACGTGTTATT GTGACTGGCAGCTTGATCCAAGCCTATCCCGCATCGCAATGCACTATTACACCCAACACTGTGTACTGTAGGTATAA TGATGCCCAAGTACTCTCAGATGATACGATGGCTTGCCTCCAAGGTAACTTGACAAGATGCACCTTCTCTCCAGTGG TTGGGAGCTTTCTCACTCGATTCGTGCTGTTCGATGGAATAGTTTATGCAAATTGCAGGTCGATGTTGTGCAAGTGC ATGCAGCCTGCTGCTGTGATCCTACAGCCGAGTTCATCCCCTGTAACTGTCATTGACATGTACAAATGTGTGAGTCT GCAGCTTGACAATCTCAGATTCACCATCACTCAATTGGCCAATGTAACCTACAATAGCACCATCAAGCTTGAAACAT CCCAGATCTTGCCTATTGATCCGTTGGATATATCCCAGAATCTAGCTGCGGTGAATAAGAGTCTAAGTGATGCACTA CAACACTTAGCACAAAGTGACACATACCTTTCTGCAATCACATCAGCTACGACTACAAGTGTATTATCCATAATAGC AATCTGTCTTGGATCGTTAGGTTTAATATTAATAATCTTGCTCAGTGTAGTTGTGTGGAAGTTATTGACCATTGTCG CTGCTAATCGAAATAGAATGGAGAATTTTGTTTATCATAATTCAGCATTCCACCACTCACGATCTGATCTCAGTGAG AAAAATCAACCTGCAACTCTTGGAACAAGATAAGACAGTCATCCATTAGTAATTTTTAAGAAAAAAACGATAGGACC GAAACTAGTATTGAAAGAACCGTCTCGGTCAATCTAGGTAATCGAGCTGCTACCGTCTCGGAAAGCTCAAATCATGC TGCCTGATCCGGAAGATCCGGAAAGCAAAAAAGCTACAAGGAGAACAGGAAACCTAATTATCTGCTTCCTATTCATC TTCTTTCCGTTTGTAAACTTCATTGTTCCAACTCTAAGACACTTGCTGTCCTAACACCTGCTATAGGCTATCCACTG CATCATCTCTTTTTAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGGTGCCAACA

[0150]

[0151] GAGCCCAGCCTGATGATCGACGGCATCCTGTGGGAGGGCTTCGGAGGCGATCCTTGTGATCCCTGCAC CACATGGTGCGACGCCATCAGCCTTAGACTGGGCTACTACGGCGACTTCGTGTTTGATAGAGTGCTGAAAACCGAT GTCAACAAGCAGTTTGAGATGGGCGCCGCTCCAACAGGCGACGCCGACCTGACCACCGCCCCTACACCAGCCAGCC GGGAAAACCCTGCCTACGGCAAGCACATGCAGGACGCCGAGATGTTTACAAACGCCGCTTATATGGCCCTGAATAT CTGGGACCGGTTCGACGTGTTCTGCACCCTGGGCGCTACATCTGGATACCTGAAGGGCAACAGCGCCGCTTTCAAC CTGGTGGGCCTGTTCGGCAGAGATGAGACGGCAGTTGCTGCTGATGACATCCCTAACGTGAGCCTGAGCCAGGCCG TTGTGGAACTGTACACCGACACCGCCTTTGCCTGGAGCGTGGGAGCCAGAGCCGCCCTGTGGGAGTGCGGTTGTGC CACACTGGGCGCCAGCTTCCAGTACGCCCAGAGCAAGCCCAAAGTGGAAGAGCTGAATGTGCTGTGCAACGCCGCA GAGTTCACCATCAACAAGCCTAAGGGCTATGTGGGCCAGGAGTTCCCCCTGAACATCAAGGCCGGCACCGTGTCCG CCACCGACACCAAGGACGCCAGCATCGATTACCACGAGTGGCAGGCCTCTCTGGCCCTCAGCTACAGACTGAACAT GTTCACACCTTACATCGGCGTGAAGTGGTCCCGCGCCAGCTTCGACGCTGATACAATCAGAATCGCCCAACCTAAG CTGGAAACCTCTATTCTGAAGATGACTACCTGGAACCCCACCATCAGCGGCAGCGGCATCGACGTGGACACCAAGA TCACCGACACCCTGCAGATCGTGTCCCTGCAACTGAACAAGATGAAAAGCAGAAAGAGCTGCGGCCTGGCTATTGG AACAACCATCGTGGACGCCGATAAGTACGCCGTGACCGTGGAAACAAGACTGATCGACGAGCGGGCCGCCCACGTG AACGCCCAGTTCCGGTTCTGAGCTAGCCTCCTGCCATACTTCCTACTCACATCATATCTATTTTAAAGAAAAAATA GGCCCGAACACTAATCGTGCCGGCAGTGCCACTGCACACACAACACTACACATACAATACACTACAATGGTTGCAG AAGATGCCCCTGTTAGGGGCACTTGCCGAGTATTATTTCGAACAACAACTTTAATTTTTCTATGCACACTATTAGC ATTAAGCATCTCTATCCTTTATGAGAGTTTAATAACCCAAAAGCAAATCATGAGCCAAGCAGGCTCAACTGGATCT AATTCTAGATTAGGAAGTATCACTGATCTTCTTAATAATATTCTCTCTGTCGCAAATCAGATTATATATAACTCTG CAGTCGCTCTACCTCTACAATTGGACACTCTTGAATCAACACTCCTTACAGCCATTAAGTCTCTTCAAACAAGTGA CAAGCTAGAACAGAACTGCTCGTGGGGTGCTGCACTGATTAATGATAATAGATACATTAATGGCATCAATCAGTTC TATTTCTCAATTGCTGAGGGTCGCAAGCTGACACTTGGCCCACTTCTTAATATACCTAGTTTCATTCCAACTGCCA CGACACCAGAGGGCTGCACCAGGATCCCATCATTCTCGCTCACCAAGACACACTGGTGTTATACACACAATGTTAT CCTGAATGGATGCCAGGATCATGTATCCTCAAATCAATTTGTTTCCATGGGAATCATTGAACCCACTTCTGCCGGG TTTCCATCCTTTCGAACCCTAAAGACTCTATATCTCAGCGATGGGGTCAATCGTAAGAGCTGCTCTATCAGTACAG TTCCGGGGGGTTGTATGATGTACTGTTTCGTCTCTACTCAACCAGAGAGGGATGACTACTTTTCTACCGCTCCTCC AGAACAACGAATTATTATAATGTACTATAATGATACAATCGTGGAGCGCATAATTAATCCACCCGGGGTACTAGAT GTATGGGCAACATTGAACCCAGGAACAGGAAGCGGGGTATATTATTTAGGTTGGGTACTCTTTCCAATATATGGCG GCGTGATTAAAGATACGAGTTTATGGAATAATCAAGCAAATAAATACTTTATCCCCCAGATGGTTGCTGCTCTCTG CTCACAAAACCAGGCAACTCAAGTCCAAAATGCTAAGTCATCATACTATAGCAGCTGGTTTGGCAATCGAATGATT CAGTCTGGGATCCTGGCATGTCCTCTTCAACAGGATCTAACCAATGAGTGTTTAGTTCTGCCCTTTTCTAATGATC AGGTGCTTATGGGTGCTGAAGGGAGATTATACATGTATGGTGACTCGGTGTATTACTACCAAAGAAGCAATAGTTG GTGGCCTATGACCATGCTGTATAAGGTAACCATAACATTCACTAATGGTCAGCCATCTGCTATATCAGCTCAGAAT GTGCCCACACAGCAGGTCCCTAGACCTGGGACAGGAAGCTGCTCTGCAAATAGATGTCCCGGTTTTTGCTTGA AAGGAGTGTATGCTGATGCCTGGTTACTGACCAACCCTTCGTCTACCAGTACATTTGGATCAGAAGCAACCTTCAC TGGTTCTTATCTCAACGCAGCAACTCAGCGTATCAATCCGACGATGTATATCGCGAACAACACACAGATCATAAGC TCACAGCAATTTGGATCAAGCGGTCAAGAAGCAGCATATGGCCCACACAACTTGTTTTTAGGGACACAGGCTCTGTTA TGGTATACTGTATCTATATTATTGAATTGTCCTCATCTCTCTTAGGACAATTTCAGATTGTCCCATTTATCCGTCA GGTGACACTATCCTAAAGGCAGAAGCCTCCAGGTCTGACCCAGCCAATCAAAGCATTATACCAGACCATGGAATGC ATACCAAACATTATTGACACTAATGACACACCAAAATTGGTTTTAAGAAAACCAAGAGAACAATAGGCCAGAATGG CTGGGTCTCGGGAGATATTACTCCCTGAAGTCCATCTCAATTCACCAATTGTAAAGCATAAGCTATACTATTACAT TCTACTTGGAAACCTCCCAAATGAGATCGACATTGACGATTTAGGTCCATTACATAATCAAAATTGGAATCAAATA GCACATGAAGAGTCTAACTTAGCCCAAGCCTTGGTAAATGTAAAAATTTCTAATTACCCACATCCCTGATCTTA GAAAGGGCATTGGCAAGAGTATGTCAATGTAATACTGTGGCCGCGAATTCTTCCCTTGATCCCGGATTTTAAAAT CAATGACCAATTGCCTCTACTCAAAAATTGGGACAAGTTAGTTAAAAGAATCATGTTCAGTAATCAATGCGGGTACT TCCCAGTGCATTCAGAATCTCAGCTATGGACTGACAGGTCGTGGGAACCTCTTTACACGATCACGTGAACTCTCTG GTGACCGCAGGGATATTGATCTTAAGACGGTTGTGGCAGCATGGCATGACTCAGACTGGAAAAGAATAAGTGATTT TTGGATTATGATCAAATTCCAGATGAGACAATTAATTGTTAGGCAAACAGATCATAATGATCCTGATTTAATCACG TATATCGAAAATAGAGAAGGCATAATCATCATAACCCCTGAACTGGTAGCATTATTTAACACTGAGAATCATACAC TAACATACATGACCTTTGAAATTGTACTGATGGTTTCAGATATGTACGAAGGTCGTCACAACATTTTATCACTATG CACAGTTAGCACTTACCTGAATCCTCTGAAGAAAAGAATAACATATTTATTGAGCCTTGTAGATAACTTAGCTTTT CAGATAGGTGATGCTGTATATAACATAATTGCTTTGCTAGAATCCTTTGTATATGCACAGTTGCAAATGTCAGATC CCATCCCAGAACTCAGAGGACAATTCCATGCATTCGTATGTTCTGAGATTCTTGATGCACTAAGGGGAACTAATAG TTTCACCCAGGATGAATCAAGAACTGTGACAACCAATTTGATATCCCCATTCCAAGATCTGACCCCAGATCTTACG GCTGAATTGCTCTGTATAATGAGGCTTTGGGGACACCCCATGCTCACCGCCAGTCAAGCTGCGGGAAAGGTACGCG AGTCCATGTGTGCTGGAAAAGTATTAGACTTTCCCACCATTATGAAAACACTAGCCTTTTTCCATACTATTCTGAT CAATGGATACAGGAGGAAGCATCATGGAGTATGGCCACCCTTAAACTTACCGGGTAATGCTTCAAAGGGTCTCACG GAACTTATGAATGACAATACTGAGATAAGCTATGAATTCACACTTAAGCATTGGAAGGAAATCTCTCTTATAAAAT TCAAGAAATGTTTTGATGCAGACGCAGGTGAGGAACTCAGTATATTTATGAAAGATAAAGCAATTAGTGCCCCAAA ACAAGATTGGATGAGTGTGTTTAGAAGAAGCCTAATCAAACAGCGCCATCAGCATCATCAGGTCCCCCTACCAAAT CCATTCAATCGACGGCTATTGCTAAACTTTCTCGGAGATGACAAATTCGACCCGAATGTGGAGCTACAGTATGTAA CATCAGGTGAGTATCTACATGATGACACGTTTTGTGCATCATATTCACTAAAAGAGAAGGAAATTAAACCTGATGG TCGAATTTTTGCAAAGTTGACTAAGAGAATGAGATCATGTCAAGTTATAGCAGAATCTCTTTTAGCGAACCATGCT GGGAAGTTAATGAAAGAGAATGGTGTTGTGATGAATCAGCTATCATTAACAAAATCACTATTAACAATGAGTCAGA TTGGAATAATATCCGAGAAAGCTAGAAAATCGACTCGAGATAACATAAATCAACCTGGTTTCCAGAATATCCAGAG AAATAAATCACATCACTCCAAGCAAGTCAATCAGCGAGATCCAAGTGATGACTTTGAATTGGCAGCATCTTTTTTA ACTACTGATCTCAAAATATTGTTTACAATGGAGGTACCAGACAATTATCCCATTTGCTCAATCATTAAACAGAA TGTATGTGTTATCCTCATCTCTTTGAGTGGATTCACTTACGGCTAATGCGTAGTACACTTTACGTGGGGGATCCCTT CAACCCACCAGCAGATACCAGTCAATTTGATCTAGATAAAGTAATTAATGGAGATATCTTCATTGTATCACCCAGA GGTGGAATTGAAGGGCTATGTCAAAAGGCTTGGACAATGATATCTATCTCTGTGATAATTCTATCTGCCACAGAGT CTGGCACACGAGTAATGAGTATGGTGCAGGGAGATAATCAAGCAATTGCGTGTCACCACACGAGTACCAAGGAGCCT GCGACTCTTGAGAAAAAGACTATTGCTTTTTAGATCTTGTAATCTATTCTTTGAGAGGTTAAAATGTAATAATTTT GGATTAGGTCACCATTTGAAAGAACAAGAGACTATCATTAGTTCTCACTTCTTTGTTTATAGCAAGAGAATATTCT ATCAGGGGAGGATTCTAACGCAAGCCTTAAAAAATGCTAGTAAGCTCTGCTTGACAGCTGATGTCCTAGGAGAATG TACCCAATCATCATGTTCTAATCTTGCAACTACTGTCATGAGGTTAACTGAGAATGGTGTTGAAAAAGATATCTGT TTCTACTTGAATATCTATATGACCATCAAACAGCTCTCCTATGATATCATCTTCCCTCAAGTGTCAATTCCTGGAG ATCAGATCACATTAGAATACATAAATAATCCACACCTGGTATCACGATTGGCTCTTCTGCCATCCCAGCTAGGAGG TCTAAACTACCTGTCATGCAGTAGGCTGTTCAATCGAAACATAGGCGACCCGGTGGTTTCCGCAGTTGCAGATCTT AAGAGATTAATTAAATCAGGATGTATGGATTACTGGATCCTTTTAACTTATTAGGAGAAAACCGGGAAACGGCT CATGGGCTACTTTAGCAGCTGACCCGTACTCAATCAATAGAGTATCAATACCCCCCAACTACAGCTCTTAAGAG GCACACCCAACAAGCTCTGATGGAACTCAGTACGAATCCAATGTTACGTGGCATATTCTCTGACAATGCACAGGCA GAAGAAAATAATCTTGCTAGATTTCTCCTGGATAGGGAGGTGATCTTTCCGCGTGTAGCTCACATCATCATTGAGC AAACCAGTGTCGGGAGGAGAAAACAGATTCAAGGATATTTGGATTCAACTAGATCGATAATGAGTAAATCACTAGA AATTAAGCCCTTGTCCAATAGGAAGCTTAATGAAATACTGGATTACACATCAATTACTAGCTTACAATTTGGCA TTACTCAAGAATGCTATTGAACCTCCGACTTATTTGAAAGCAATGACTCTTGAAACATGTAGCATCGACATTGCAA GGAGCCTCCGGAAGCTCTCCTGGGCCCCACTCTTGGGTGGGAGAAATCTTGAAGGATTAGAGACGCCAGATCCCAT TGAAATTACTGCAGGAGCATTAATTGTTGGATCGGGCTACTGTGAACAGTGTGCTGCAGGAGACAATCGATTCACA TGGTTTTTCTTGCCATCTGGTATCGAGATAGGAGGGGATCCCCGTGATAATCCTCCTATCCGTGTACCGTACATTG GCTCCAGGACTGATGAGAGGAGGGTAGCCTCAATGGCATACATCAGGGGTGCCTCGAGTAGCCTAAAAGCAGTTCT TAGACTGGCGGGAGTGTACATCTGGGCATTCGGAGATACTCTGGAGAATTGGATAGATGCACTGGATTTGTCTCAC ACTAGAGTTAACATCACACTTGAACAGCTGCAATCCCTCACCCCACTTCCAACCTCTGCCAATCTAACCCATCGGT TGGATGATGGCACAACTACCCTAAAGTTTACTCCTGCGAGCTCTTACACCTTTTCAAGTTTCACTCATATATCAAA TGATGAGCAATACCTGACAATTAATGACAAAACTGCAGATTCAAATATAATCTACCAACAGTTAATGATCACTGGA CTCGGAATTTTAGAAACATGGAATAATCCCCCAATCAATAGAACATTCGAAGAATCTACCCTACATTTGCACACTG GTGCATCATGTTGTGTCCGACCTGTGGACTCCTGCATCATCTCAGAAGCATTAACAGTCAAGCCACATATTACAGT ACCGTACAGCAATAAATTTGTATTTGATGAAGACCCGCTATCTGAATATGAGACTGCAAAACTGGAATCGTTATCA TTTCAAGCCCAATTAGGCAACATTGATGCTGTAGATATGACAGGTAAATTAACATTATTGTCCCAATTCACTGCAA GGCAGATTATTAATGCAATCACTGGACTCGATGAGTCTGTCTCTCTTACTAATGATGCCATTGTTGCATCAGACTA TGTCTCCAATTGGATTAGTGAATGCATGTATACCAAATTAGATGAATTATTTATGTATTGTGGGTGGGAACTACTA TTGGAACTATCCTATCAAATGTATTATCTGAGGGTAGTTGGGTGGAGTAATATAGTGGATTATTCTTACATGATCT TGAGAAGAATCCCGGGTGCAGCATTAAACAATCTGGCATCTACATTAAGTCATCCAAAACTTTTCCGACGAGCTAT CAACCTAGATATAGTTGCCCCCTTAAATGCTCCTCATTTTGCATCTCTGGACTACATCAAGATGAGTGTGGATGCA ATACTCTGGGGCTGTAAAAGTCATCAATGGTTCTCCCAATGGAGGGGACTTAATTAGTTGTGACATCTGAAG ATAGCCTTATTCTCAGTGACCGATCCATGAATCTCATTGCAAGGAAATTAACTTTATTATCACTGATTCACCATAA TGGTTTGGAACTACCAAAGATTEAAGGGGGTTCTCTCCTGATGAGAAGTGTTTCGCTTTGACAGAATTTTTGAGGAAA GTGGTGAACTCAGGGTTGAGTTCAATAGAGAACCTATTCAAATTTTATGTACAATGTGGAGAACCCACGGCTTGCAG CATTCGCCAGCAACAATTACTACCTGACCAGAAAATTATTGAATTCAATACGAGATACTGAGTCGGGTCAAGTAGC AGTCACCTCATATTATGAATCATTAGATATATTGATAGTCTTAAGCTAACCCCACATGTGTGCCTGGTACCTCATGC ATTGAGGATGATAGTCTATGTACAAATGATTACATAATCTGGATCATAGAGTCTAATGCAAACTTGGAGAAGTATC CAATTCCAAATAGCCCTGAGGATGATTCCAATTTCCATAACTTTAAGTTGAATGCTCCATCGCACCATACCTTACG CCCATTAGGGTTGTCATCAACTGCTTGGTATAAGGGTATAAGCTGTTGCAGGTACCTTGAGCGATTAAAGCTACCA CAAGGTGATCATTTATATTGCAGAAGGTAGTGGTGCCAGTATGACAATCATAGAATACCTATTCCAGGAAGAA AGATATATTACAATTCTTTATTTAGGTGGTGACAATCCCCCACAAAGAAATTATGCACCAATGCCTACTCAGTT CATTGAGAGTGTCCCATACAAGCTCTGGCAAGCACACAGATCAATATCCCGAGATTTTTGAGGACTTCATCCCT CTATGGAACGGAAATGCCGCCATGACTGACATAGGAATGACAGCTTGTGTAGAATTCATCATCAATCGAGTCGGCC CAAGGACTTGCAGTTTAGTACATGTAGATTTGGAATCAAGTGCAAGCTTAAATCAACAATGCCTGTCAAAGCCGAT AATTAATGCTATCATCACTGCTACAACTGTTTTGTGCCCTCATGGGGTGCTTATTCTGAAATATAGTTGGTTGCCA TTTACTAGATTTAGTACTTTGATCACTTTCTTATGGTGCTACTTTGAGAGAATCACTGTTCTTAGGAGCACATATT CTGGTCCAGCTAATCATGAGGTTTATTTAATTTGTATCCTTGCCAACAACTTTGCATTCCAGACTGTCTCGCAGGC AACAGGAATGGCGATGACTTTTAACCGATCAAGGGTTTACTTTGATATCACCTGAAAGAATAAATCAGTATTGGGAT GGTCACTTGAAGCAAGAACGTATCGTAGCAGAAGCAATTGATAAGGTGGTTCTAGGAGAAGATGCTCTATTCAATT CGAGTGATAATGAATTAATTCTCAAATGTGGAGGGACACCAAATGCACGGAATCTTATCGATATCGAGCCAGTCGC AACTTTCATAGAATTTGAACAACTGATCTGCACAATGTTGACAACCCACTTGAAGGAAATAATTGATATAACAAGG TCTGGAACCCAGGATTATGAAAGTTTATTACTCACTCCTTACAATTTAGGTCTTCTTGGTAAAATCAGTACGATAG TGAGATTATTAACAGAAAGGATTCTAAATCATACTATCAGGAATTGGTTGATCCTCCCACCTTCGCTCCGGATGAT CGTGAAGCAGGACTTGGAATTCGGCATATTCAGGATTACTTCCATCCTCAATTCTGATCGGTTCCTGAAGCTTTCT CCAAATAGGAAATACTTGATTACACAATTAACTGCAGGCTACATTAGGAAATTGATTGAGGGGGATTGTAATATCG ATCTAACCAGACCTATCCAAAAACAAATCTGGAAAGCATTAGGTTGTGTAGTCTATTGTCACGATCCAGTAGATCA AAGGGAATCAACAGAGTTTATTGATATAAATATTAATGAAGAAATAGACCGCGGGATCGATGGCGAGGAAATCTAA ATATATCAAGAATCAGAATTAGTTTAAGAAAAAAGAAGAGGATTAATCTTGGTTTTCCCCTTGGT

[0152] The underlined sequence is the PIV5-CPI genomic nucleic acid sequence. The italicized and underlined sequences are the entire MOMP nucleic acid sequence. The double-underlined sequence is the AOS-modified MOMPΔsp-CD5sp nucleic acid sequence.

[0153] c. pMHD177- CPI-NT-MOMPΔsp

[0154] The pMHD177-CPI-NT-MOMPΔsp nucleic acid sequence provided in this article, from 5' to 3', is as follows:

[0155] ACCAGGGGGAAAACGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCTA TGGCCTTCGTGACCGACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCGACA CAAAAATGTCATCCGTGCTTAAAGCATATGAGCGATTCACACTCACTCAAGAACTGCAAGATCAGAGTGAGGAAGGT ACAATCCCACCTACAACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGATCCCG GCTTCTTCTATTCTGCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACTTACAA TGTTTTCGCTACCATCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATCGAAAGG GTAGAGATCGATGGCTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCTCGTTCAGGTATGAGCCGTGGAGAGAT CAATGCCTATGCTGCACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTTGTTGATTCCGAAGTCG AGGGAACTGCATGGGATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGGATAGTGACT TGCAAGTGCATGACTGCGCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAATATCGTCAGCAAGGCAG GATCAACCCGAGATATCTCCTGCAACCGGAGGCTCGAAGAATAATCCAGAATGTAATCCGAAAGGGAATGGTGGTCA GACATTTCCTCACCTTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTATGCTATGGTAGGG GATGTTGGAAAGTATATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAATATGCATTAGGAACCAGATG GCCCACACTTGCTTTAGCTGCATTTTCAGGAGAGCTAACAAAGCTAAAGTCCCTCATGGCATTATACCAGACCCTTG GTGAGCAGGCCCGATATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTTGCTGCAGCAAACTACCCACTGCTA TATAGCTATGCTATGGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTCTCCAGATCATACATGAA TAAGACATATTTCCAATTGGGAATGGAAACTGCAAGAAAACAACAGGGTGCAGTTGACATGAGGATGGCAGAAGATC TCGGTCTAACTCAAGCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACAGCAAATCGAGGGGCAGAC ACCAGGGGAGGAGTCAACCCGTTCTCATCTATCACTGGGACAACTCAGGTGCCCGCTGCAGCAACAGGTGACACATT CGAGAGTTACATGGCAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCACGATGATGAGATGCCACCAT TGGAAGAGGAGGAAGAGGACGACACATCTGCAGGTCCACGCACTGGACTAACTCTTGAACAAGTGGCCTTGGACATC CAGAACGCAGCAGTTGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGGGTGATCTTGACATCTAGACAAT TCAGATCCCAATCTTAAATCGACACACCTAATTGACCAGTTAGATGGAACTACAGTGGATTCCATAAGGTTCCTGCC TACCATCGGCTTTTAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGATGCCAATAACACAATCCA CAATCTACAATGGATCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATAAGCTCATAGAGACAGGCCTGAATAC TGTGGAGTATTTTACTTCCCAACAAGTCACAGGAACATCCTCTCTTGGAAAGAATACAATACCACCAGGGGTCACAG GACTACTAACCAATGCTGCAGAGGCAAAGATCCAAGAGTCAACCAACCATCAGAAGGGTTCAGTTGGTGGGGGCGCA AAACCAAAGAAACCGCGACCAAAAATTGCCATTGTGCCAGCAGATGACAAAACGGTGCCCGGAAAGCCGATCCCAAA CCCTCTATTAGGTCTGGACTCCACCCCGAGCACCCAAACTGTGCTTGATCTAAGTGGGAAAACATTACCATCAGGAT CCTATAAGGGGGTTAAACTTGCGAAATTTGGAAAAGAAAATCTGATGACACGGTTCATCGAGGAACCCAGAGAGAAT CCTATCGCAACCAATTCCCCCATCGATTTTAAGAGGGGCAGGGATACCGGCGGGTTCCATAGAAGGGAGTACTCAAT CGGATGGGTGGGAGATGAAGTCAAGGTCACTGAGTGGTGCAATCCATCCTGTTCTCCAATCACCGCTGCAGCAAGGC GATTTGAATGCACTTGTCACCAGTGTCCAGTCACTTGCTCTGAATGTGAACGAGATACTTAATACAGTGAGAAATTT GGACTCTCGGATGAATCAACTGGAGACAAAAGTAGATCGCATTCTCTCATCTCAGTCTCTAATCCAGACCATCAAGA ATGACATAGTTGGACTTAAAGCAGGGATGGCTACTTTAGAAGGAATGATTACAACTGTGAAAATCATGGACCCGGGA GTTCCCAGTAATGTTACTGTGGAAGATGTACGCAAGAAACTAAGTAACCATGCTGTTGTTGTGCCAGAATCATTCAA TGATAGTTTCTTGACTCAATCTGAAGATGTAATTTCACTTGATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAGA AGATTGTCAGGAAGGTTCCTCCTCAGAAGGATCTGACTGGATTGAAGATCACACTAGAGCAATTGGCAAAGGATTGC ATCAGCAAACCGAAGATGAGGGAAGAGTATCTCCTCAAAATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCT CAAGAAAGCAATCATCCGCAGTGCAATTTGATCAAGAAACACCCAATTACACTACACTGGTATGACACTGTACTAAC CCTGAGGGTTTTAGAAAAAACGATTAACGATAAATAAGCCCGAACACTACACACCACCCGAGGCAGCCATGCCATCC ATCAGCATCCCCGCAGACCCCACCAATCCACGTCAATCAATAAAAGCGTTCCCAATTGTGATCAACCGTGATGGGGG TGAGAAAGGTCGCTTGGTTAAACAACTACGCACAACCTACTTGAATGACCTAGATACTCATGAGCCACTGGTGACAT TCGTAAATACTTATGGATTCATCTACGAACAGGATCGGGGAAATACCATTGTCGGAGAGGATCAACATGGGAAGAAA AGAGAGGCTGTGACTGCTGCAATGGTTACCCTTGGATGTGGGCCTAATCTACCATCATTAGGGAATGTCCTGGGACA ACTGAGTGAATTCCAGGTCATTGTTAGGAAGACATCCAGCAAAGCGGAAGAGATGGTCTTTGAAATTGTTAAGTATC CGAGAATATTTCGGGGTCATACATTAATCCAGAAAGGACTAGTCTGTGTCTCCGCAGAAAAATTTGTTAAGTCACCA GGGAAAGTACAATCTGGAATGGACTATCTCTTCATTCCGACATTTCTGTCAGTGACTTACTGTCCAGCTGCAATCAA ATTCAGGTACCTGGCCCCATGTTGAAAATGAGATCAAGATACACTCAGAGCTTACAACTTGAACTAATGATAAGAA TCCTGTGTAAGCCCGATTCGCCACTTATGAAGGTCCATATCCCTGACAAGGAAGGAAGAGGATGTCTTGTATCAGTA TGGTTGCATGTATGCAATATCTTCAAATCAGGAAACAAGAATGGCAGTGAGTGGCAGGAATACTGGATGAGAAAGTG TGCTAACATGCAACTTGAAGTGTCGATTGCAGATATGTGGGGACCAACTATCATAATTCATGCCAGAGGTCACATTC CCAAAAGTGCTAAGTTGTTTTTTGGAAAGGGTGGATGGAGCTGCCATCCACTTCACGAAGTTGTTCCAAGTGTCACT AAAACACTATGGTCCGTGGGCTGTGAGATTACAAAGGCGAAGGCAATAATACAAGAGAGTAGCATCTCTCTTCTCGT GGAGACTACTGACATCATAAGTCCAAAAGTCAAAATTTCATCTAAGCATCGCCGCTTTGGGAAATCAAATTGGGGTC TGTTCAAGAAAACCAAATCACTGCCCAACCTGACGGAGCTGGAATGACTGACCCCCAATCGAGACTACACCACCTCA AACTATAGGTGGGTGGTACCTCAGTGATTAATCTCGTAAGCACTGATCGTAGGCTACAACACACTAATATTATCCAG ATTAGAGAGCTTAATTAGCTCTGTATTAATAATAACACTACTATTCCAATAACTGGAATCACCAGCTTGATTTATCT CCAAAATGATTCAAAGAAAACAAATCATATTAAGACTATCCTAAGCACGAACCCATATCGTCCTTCAAATCATGGGT ACTATAATTCAATTTCTGGTGGTCTCCTGTCTATTGGCAGGAGCAGGCAGTCTTGATCCAGCAGCCCTCATGCAAAT CGGTGTCATTCCAACAAATGTCCGGCAACTTATGTATTATACTGAGGCCTCATCAGCATTCATTGTTGTGAAGTTAA TGCCTACAATTGACTCGCCGATTAGTGGATGTAATATAACATCAATTTCAAGCTATAATGCAACAGTGACAAAACTC CTACAGCCGATCGGTGAGAATTTGGAGACGATTAGGAACCAGTTGATTCCAACTCGGAGGAGACGCCGGTTTGCAGG GGTGGTGATTGGATTAGCTGCATTAGGAGTAGCTACTGCCGCACAGGTCACTGCCGCAGTAGCACTAGTAAAGGCAA ATGAAAATGCTGCGGCTATACTCAATCTCAAAAATGCAATCCAAAAAACAAATGCAGCAGTTGCAGATGTGGTCCAG GCCACACAATCACTAGGAACAGCAGTTCAAGCAGTTCAAGATCACATAAACAGTGTGGTAAGTCCAGCAATTACAGC AGCCAATTGTAAAGCCCAAGATGCTATCATTGGCTCAATCCTCAATCTCTATTTGACCGAGTTGACAACTATCTTCC ACAATCAAATTACAAACCCTGCATTGAGTCCTATTACAATTCAAGCTTTAAGGATCCTACTAGGGAGTACCTTGCCG ACTGTGGTCGAAAAATCTTTCAATACCCAGATAAGTGCGGCTGAGCTTCTCTCATCAGGGTTATTGACAGGCCAGAT TGTGGGATTAGATTTGACCTATATGCAGATGGTCATAAAAATTGAGCTGCCAACTTTAACTGTACAACCTGCAACCC AGATCATAGATCTGGCCACCATTTCTGCATTCATTAACAATCAAGAAGTCATGGCCCAATTACCAACACGTGTTATT GTGACTGGCAGCTTGATCCAAGCCTATCCCGCATCGCAATGCACTATTACACCCAACACTGTGTACTGTAGGTATAA TGATGCCCAAGTACTCTCAGATGATACGATGGCTTGCCTCCAAGGTAACTTGACAAGATGCACCTTCTCTCCAGTGG TTGGGAGCTTTCTCACTCGATTCGTGCTGTTCGATGGAATAGTTTATGCAAATTGCAGGTCGATGTTGTGCAAGTGC ATGCAGCCTGCTGCTGTGATCCTACAGCCGAGTTCATCCCCTGTAACTGTCATTGACATGTACAAATGTGTGAGTCT GCAGCTTGACAATCTCAGATTCACCATCACTCAATTGGCCAATGTAACCTACAATAGCACCATCAAGCTTGAAACAT CCCAGATCTTGCCTATTGATCCGTTGGATATATCCCAGAATCTAGCTGCGGTGAATAAGAGTCTAAGTGATGCACTA CAACACTTAGCACAAAGTGACACATACCTTTCTGCAATCACATCAGCTACGACTACAAGTGTATTATCCATAATAGC AATCTGTCTTGGATCGTTAGGTTTAATATTAATAATCTTGCTCAGTGTAGTTGTGTGGAAGTTATTGACCATTGTCG CTGCTAATCGAAATAGAATGGAGAATTTTGTTTATCATAATTCAGCATTCCACCACTCACGATCTGATCTCAGTGAG AAAAATCAACCTGCAACTCTTGGAACAAGATAAGACAGTCATCCATTAGTAATTTTTAAGAAAAAAACGATAGGACC GAAACTAGTATTGAAAGAACCGTCTCGGTCAATCTAGGTAATCGAGCTGCTACCGTCTCGGAAAGCTCAAATCATGC TGCCTGATCCGGAAGATCCGGAAAGCAAAAAAGCTACAAGGAGAACAGGAAACCTAATTATCTGCTTCCTATTCATC TTCTTTCCGTTTGTAAACTTCATTGTTCCAACTCTAAGACACTTGCTGTCCTAACACCTGCTATAGGCTATCCACTG CATCATCTCTTTTTAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGGTGCCAACA

[0156]

[0157] GCATCCTGTGGGAGGGCTTCGGAGGCGATCCTTGTGATCCCTGCACCACATGGTGCGACGCCATCAGC CTTAGACTGGGCTACTACGGCGACTTCGTGTTTGATAGAGTGCTGAAAACCGATGTCAACAAGCAGTTTGAGATGG GCGCCGCTCCAACAGGCGACGCCGACCTGACCACCGCCCCTACACCAGCCAGCCGGGAAAACCCTGCCTACGGCAA GCACATGCAGGACGCCGAGATGTTTACAAACGCCGCTTATATGGCCCTGAATATCTGGGACCGGTTCGACGTGTTC TGCACCCTGGGCGCTACATCTGGATACCTGAAGGGCAACAGCGCCGCTTTCAACCTGGTGGGCCTGTTCGGCAGAG ATGAGACGGCAGTTGCTGCTGATGACATCCCTAACGTGAGCCTGAGCCAGGCCGTTGTGGAACTGTACACCGACAC CGCCTTTGCCTGGAGCGTGGGAGCCAGAGCCGCCCTGTGGGAGTGCGGTTGTGCCACACTGGGCGCCAGCTTCCAG TACGCCCAGAGCAAGCCCAAAGTGGAAGAGCTGAATGTGCTGTGCAACGCCGCAGAGTTCACCATCAACAAGCCTA AGGGCTATGTGGGCCAGGAGTTCCCCCTGAACATCAAGGCCGGCACCGTGTCCGCCACCGACACCAAGGACGCCAG CATCGATTACCACGAGTGGCAGGCCTCTCTGGCCCTCAGCTACAGACTGAACATGTTCACACCTTACATCGGCGTG AAGTGGTCCCGCGCCAGCTTCGACGCTGATACAATCAGAATCGCCCAACCTAAGCTGGAAACCTCTATTCTGAAGA TGACTACCTGGAACCCCACCATCAGCGGCAGCGGCATCGACGTGGACACCAAGATCACCGACACCCTGCAGATCGT GTCCCTGCAACTGAACAAGATGAAAAGCAGAAAGAGCTGCGGCCTGGCTATTGGAACAACCATCGTGGACGCCGAT AAGTACGCCGTGACCGTGGAAACAAGACTGATCGACGAGCGGGCCGCCCACGTGAACGCCCAGTTCCGGTTCTGAT GAGCTAGCCTCCTGCCATACTTCCTACTCACATCATATCTATTTTAAAGAAAAAATAGGCCCGAACACTAATCGTG CCGGCAGTGCCACTGCACACACAACACTACACATACAATACACTACAATGGTTGCAGAAGATGCCCCTGTTAGGGG CACTTGCCGAGTATTATTTCGAACAACAACTTTAATTTTTCTATGCACACTATTAGCATTAAGCATCTCTATCCTT TATGAGAGTTTAATAACCCAAAAGCAAATCATGAGCCAAGCAGGCTCAACTGGATCTAATTCTAGATTAGGAAGTA TCACTGATCTTCTTAATAATATTCTCTCTGTCGCAAATCAGATTATATATAACTCTGCAGTCGCTCTACCTCTACA ATTGGACACTCTTGAATCAACACTCCTTACAGCCATTAAGTCTCTTCAAACAAGTGACAAGCTAGAACAGAACTGC TCGTGGGGTGCTGCACTGATTAATGATAATAGATACATTAATGGCATCAATCAGTTCTATTTCTCAATTGCTGAGG GTCGCAAGCTGACACTTGGCCCACTTCTTAATATACCTAGTTTCATTCCAACTGCCACGACACCAGAGGGCTGCAC CAGGATCCCATCATTCTCGCTCACCAAGACACACTGGTGTTATACACACAATGTTATCCTGAATGGATGCCAGGAT CATGTATCCTCAAATCAATTTGTTTCCATGGGAATCATTGAACCCACTTCTGCCGGGTTTCCATCCTTTCGAACCC TAAAGACTCTATATCTCAGCGATGGGGTCAATCGTAAGAGCTGCTCTATCAGTACAGTTCCGGGGGGTTGTATGAT GTACTGTTTCGTCTCTACTCAACCAGAGAGGGATGACTACTTTTCTACCGCTCCTCCAGAACAACGAATTATTATA ATGTACTATAATGATACAATCGTGGAGCGCATAATTAATCCACCCGGGGTACTAGATGTATGGGCAACATTGAACC CAGGAACAGGAAGCGGGGTATATTATTTAGGTTGGGTACTCTTTCCAATATATGGCGGCGTGATTAAAGATACGAG TTTATGGAATAATCAAGCAAATAAATACTTTATCCCCCAGATGGTTGCTGCTCTCTGCTCACAAAACCAGGCAACT CAAGTCCAAAATGCTAAGTCATCATACTATAGCAGCTGGTTTGGCAATCGAATGATTCAGTCTGGGATCCTGGCAT GTCCTCTTCAACAGGATCTAACCAATGAGTGTTTAGTTCTGCCCTTTTCTAATGATCAGGTGCTTATGGGTGCTGA AGGGAGATTATACATGTATGGTGACTCGGTGTATTACTACCAAAGAAGCAATAGTTGGTGGCCTATGACCATGCTG TATAAGGTAACCATAACATTCACTAATGGTCAGCCATCTGCTATATCAGCTCAGAATGTGCCCACACAGCAGGTCC CTAGACCTGGGACAGGAAGCTGCTCTGCAACAAATAGATGTCCCGGTTTTTGCTTGAAAGGAGTGTATGCTGATGC CTGGTTACTGACCAACCCTTCGTCTACCAGTACATTTGGATCAGAAGCAACCTTCACTGGTTCTTATCTCAACGCA GCAACTCAGCGTATCAATCCGACGATGTATATCGCGAACAACACACAGATCATAAGCTCACAGCAATTTGGATCAA GCGGTCAAGAAGCAGCATATGGCCACACAACTTGTTTTAGGGACACAGGCTCTGTTATGGTATACTGTATCTATAT TATTGAATTGTCCTCATCTCTCTTAGGACAATTTCAGATTGTCCCATTTATCCGTCAGGTGACACTATCCTAAAGG CAGAAGCCTCCAGGTCTGACCCAGCCAATCAAAGCATTATACCAGACCATGGAATGCATACCAAACATTATTGACA CTAATGACACACAAAATTGGTTTTAAGAAAAACCAAGAGAACAATAGGCCAGAATGGCTGGGTCTCGGGAGATATT ACTCCCTGAAGTCCATCTCAATTCACCAATTGTAAAGCATAAGCTATACTATTACATTCTACTTGGAAACCTCCCA AATGAGATCGACATTGACGATTTAGGTCCATTACATAATCAAAATTGGAATCAAATAGCACATGAAGAGTCTAACT TAGCCCAACGCTTGGTAAATGTAAGAAATTTTCTAATTACCCACATCCCTGATCTTAGAAAGGGCCATTGGCAAGA GTATGTCAATGTAATACTGTGGCCGCGAATTCTTCCCTTGATCCCGGATTTTAAAATCAATGACCAATTGCCTCTA CTCAAAAATTGGGACAAGTTAGTTAAAGAATCATGTTCAGTAATCAATGCGGGTACTTCCCAGTGCATTCAGAATC TCAGCTATGGACTGACAGGTCGTGGGAACCTCTTTACACGATCACGTGAACTCTCTGGTGACCGCAGGGATATTGA TCTTAAGACGGTTGTGGCAGCATGGCATGACTCAGACTGGAAAAGAATAAGTGATTTTTGGATTATGATCAAATTC CAGATGAGACAATTAATTGTTAGGCAAACAGATCATAATGATCCTGATTTAATCACGTATATCGAAAATAGAGAAG GCATAATCATCATAACCCCTGAACTGGTAGCATTATTTAACACTGAGAATCATACACTAACATACATGACCTTTGA AATTGTACTGATGGTTTCAGATATGTACGAAGGTCGTCACAACATTTTATCACTATGCACAGTTAGCACTTACCTG AATCCTCTGAAGAAAAGAATAACATATTTATTGAGCCTTGTAGATAACTTAGCTTTTCAGATAGGTGATGCTGTAT ATAACATAATTGCTTTGCTAGAATCCTTTGTATATGCACAGTTGCAAATGTCAGATCCCATCCCAGAACTCAGAGG ACAATTCCATGCATTCGTATGTTCTGAGATTCTTGATGCACTAAGGGGAACTAATAGTTTCACCCAGGATGAATCA AGAACTGTGACAACCAATTTGATATCCCCATTCCAAGATCTGACCCCAGATCTTACGGCTGAATTGCTCTGTATAA TGAGGCTTTGGGGACACCCCATGCTCACCGCCAGTCAAGCTGCGGGAAAGGTACGCGAGTCCATGTGTGCTGGAAA AGTATTAGACTTTCCCACCATTATGAAAACACTAGCCTTTTTCCATACTATTCTGATCAATGGATACAGGAGGAAG CATCATGGAGTATGGCCACCCTTAAACTTACCGGGTAATGCTTCAAAGGGTCTCACGGAACTTATGAATGACAATA CTGAGATAAGCTATGAATTCACACTTAAGCATTGGAAGGAAATCTCTCTTATAAAATTCAAGAAATGTTTTGATGC AGACGCAGGTGAGGAACTCAGTATATTTATGAAAGATAAAGCAATTAGTGCCCCAAAACAAGATTGGATGAGTGTG TTTAGAAGAAGCCTAATCAAACAGCGCCATCAGCATCATCAGGTCCCCCTACCAAATCCATTCAATCGACGGCTAT TGCTAAACTTTCTCGGAGATGACAAATTCGACCCGAATGTGGAGCTACAGTATGTAACATCAGGTGAGTATCTACA TGATGACACGTTTTGTGCATCATATTCACTAAAAGAGAAGGAAATTAAACCTGATGGTCGAATTTTTGCAAAGTTG ACTAAGAGAATGAGATCATGTCAAGTTATAGCAGAATCTCTTTTAGCGAACCATGCTGGGAAGTTAATGAAAGAGA ATGGTGTTGTGATGAATCAGCTATCATTAACAAAATCACTATTAACAATGAGTCAGATTGGAATAATATCCGAGAA AGCTAGAAAATCGACTCGAGATAACATAAATCAACCTGGTTTCCAGAATATCCAGAGAAATAAATCACATCACTCC AAGCAAGTCAATCAGCGAGATCCAAGTGATGACTTTGAATTGGCAGCATCTTTTTTAACTACTGATCTCAAAAAAT ATTGTTTACAATGGAGGTACCAGACAATTATCCCATTTGCTCAATCATTAAACAGAATGTATGGTTATTCCTCATCT CTTTGAGTGGATTCACTTACGGCTAATGCGTAGTACACTTTACGTGGGGGATCCCTTCAACCCACCAGCAGATACC AGTCAATTTGATCTAGATAAAGTAATTAATGGAGATATCTTCATTGTATCACCCAGAGGTGGAATTGAAGGGCTAT GTCAAAAGGCTTGGACAATGATATCTATCTCTGTGATAATTCTATCTGCCACAGAGTCTGGCACACGAGTAATGAG TATGGTGCAGGGAGATAATCAAGCAATTGCTGTCACCACACGAGTACCAAGGAGCCTGCCGACTCTTGAGAAAAAG ACTATTGCTTTAGATCTTGTAATCTATTCTTTGAGAGGTTAAAATGTAATAATTTTGGATTAGGTCACCATTTGA AAGAACAAGAGACTATCATTAGTTCTCACTTCTTTGTTTATAGCAAGAGAATATTCTATCAGGGGAGGATTCTAAC GCAAGCCTTAAAAAATGCTAGTAAGCTCTGCTTGACAGCTGATGTCCTAGGAGAATGTACCCAATCATCATGTTCT AATCTTGCAACTACTGTCATGAGGTTAACTGAGAATGGTGTTGAAAAAGATATCTGTTTCTACTTGAATATCTATA TGACCATCAAACAGCTCTCCTATGATATCATCTTCCCTCAAGTGTCAATTCCTGGAGATCAGATCACATTAGAATA CATAAATAATCCACACCTGGTATCACGATTGGCTCTTCTGCCATCCCAGCTAGGAGGTCTAAACTACCTGTCATGC AGTAGGCTGTTCAATCGAAACATAGGCGACCCGGTGGTTTCCGCAGTTGCAGATCTTAAGAGATTAATTAAATCAG GATGTATGGATTACTGGATCCTTTATAACTTATTAGGGAGAAAACCGGGAAACGGCTCATGGGCTACTTTAGCAGC TGACCCGTACTCAATCAATATAGAGTATCAATACCCCCCAACTACAGCTCTTAAGAGGCACACCCAACAAGCTCTG ATGGAACTCAGTACGAATCCAATGTTACGTGGCATATTCTCTGACAATGCACAGGCAGAAGAAAATAATCTTGCTA GATTTCTCCTGGATAGGGAGGTGATCTTTCCGCGTGTAGCTCACATCATCATTGAGCAAACCAGTGTCGGGAGGAG AAAACAGATTCAAGGATATTTGGATTCAACTAGATCGATAATGAGTAAATCACTAGAAATTAAGCCCTTGTCCAAT AGGAAGCTTAATGAAATACTGGATTACAACATCAATTACCTAGCTTACAATTTGGCATTACTCAAGAATGCTATTG AACCTCCGACTTATTTGAAAGCAATGACTCTTGAAACATGTAGCATCGACATTGCAAGGAGCCTCCGGAAGCTCTC CTGGGCCCCACTCTTGGGTGGGAGAAATCTTGAAGGATTAGAGACGCCAGATCCCATTGAAATTACTGCAGGAGCA TTAATTGTTGGATCGGGCTACTGTGAACAGTGTGCTGCAGGAGACAATCGATTCACATGGTTTTTCTTGCCATCTG GTATCGAGATAGGAGGGGATCCCCGTGATAATCCTCCTATCCGTGTACCGTACATTGGCTCCAGGACTGATGAGAG GAGGGTAGCCTCAATGGCATACATCAGGGGTGCCTCGAGTAGCCTAAAAGCAGTTCTTAGACTGGCGGGAGTGTAC ATCTGGGCATTCGGAGATACTCTGGAGAATTGGATAGATGCACTGGATTTGTCTCACACTAGAGTTAACATCACAC TTGAACAGCTGCAATCCCTCACCCCACTTCCAACCTCTGCCAATCTAACCCATCGGTTGGATGATGGCACAACTAC CCTAAAGTTTACTCCTGCGAGCTCTTACACCTTTTCAAGTTTCACTCATATATCAAATGATGAGCAATACCTGACA ATTAATGACAAAACTGCAGATTCAAATATAATCTACCAACAGTTAATGATCACTGGACTCGGAATTTTAGAAACAT GGAATAATCCCCCAATCAATAGAACATTCGAAGAATCTACCCTACATTTGCACACTGGTGCATCATGTTGTGTCCG ACCTGTGGACTCCTGCATCATCTCAGAAGCATTAACAGTCAAGCCACATATTACAGTACCGTACAGCAATAAATTT GTATTTGATGAAGACCCGCTATCTGAATATGAGACTGCAAAACTGGAATCGTTATCATTTCAAGCCCAATTAGGCA ACATTGATGCTGTAGATATGACAGGTAAATTAACATTATTGTCCCAATTCACTGCAAGGCAGATTATTAATGCAAT CACTGGACTCGATGAGTCTGTCTCTCTTACTAATGATGCCATTGTTGCATCAGACTATGTCTCCAATTGGATTAGT GAATGCATGTATACCAAATTAGATGAATTATTTATGTATTGTGGGTGGGAACTACTATTGGAACTATCCTATCAAA TGTATTATCTGAGGGTAGTTGGGTGGAGTAATATAGTGGATTATTCTTACATGATCTTGAGAAGAATCCCGGGTGC AGCATTAAACAATCTGGCATCTACATTAAGTCATCCAAAACTTTTCCGACGAGCTATCAACCTAGATATAGTTGCC CCCTTAAATGCTCCTCATTTTGCATCTCTGGACTACATCAAGATGAGTGTGGATGCAATACTCTGGGGCTGTAAAA GAGTCATCAATGGTTCTCCCAATGGAGGGGACTTAGAATTAGTTGTGACATCTGAAGATAGCCTTATTCTCAGTGA CCGATCCATGAATCTCATTGCAAGGAAATTAACTTTATTATCACTGATTCACCATAATGGTTTGGAACTACCAAAG ATTAAGGGGTTCTCTCCTGATGAGAAGTGTTTCGCTTTGACAGAATTTTTGAGGAAAGTGGTGAACTCAGGGTTGA GTTCAATAGAGAACCTATCAAATTTTATGTACAATGTGGAGAACCCACGGCTTGCAGCATTCGCCAGCAACAATTA CTACCTGCCAGAAAATTATTGAATTCAATACGAGATACTGAGTCGGGTCAAGTAGCAGTCACCTCATATTATGAA TCATTAGAATATATTGATAGTCTTAAGCTAACCCCACATGTGCCTGGTACCTCATGCATTGAGGATGATAGTCTAT GTACAAATGATTACATAATCTGGATCATAGAGTCTAATGCAAACTTGGAGAAGTATCCAATTCCAAATAGCCCTGA GGATGATTCCAATTTCCATAACTTTAAGTTGAATGCTCCATCGCACCATACCTTACGCCCATTAGGGTTGTCATCA ACTGCTTGGTATAAGGGTATAAGCTGTTGCAGGTACCTTGAGCGATTAAAGCTACCACAAGGTGATCATTTATATA TTGCAGAAGGTAGTGGTGCCAGTATGACAATCATAGAATACCTATTCCCAGGAAGAAAGATATATTACAATTCTTT ATTTAGTAGTGGTGACAATCCCCCACAAAGAAATTATGCACCAATGCCTACTCAGTTCATTGAGAGTGTCCCATAC AAGCTCTGGCAAGCACACACAGATCAATATCCCGAGATTTTTGAGGACTTCATCCCTCTATGGAACGGAAATGCCG CCATGACTGACATAGGAATGACAGCTTGTGTAGAATTCATCATCAATCGAGTCGGCCCAAGGACTTGCAAGTTTAGT ACATGTAGATTTGGAATCAAGTGCAAGCTTAAATCAACAATGCCTGTCAAAGCCGATAATTAATGCTATCATCACT GCTACAACTGTTTTGTGCCCTCATGGGGTGCTTATTCTGAAATATAGTTGGTTGCCATTTACTAGATTTAGTACTT TGATCACTTTTCTTATGGTGCTACTTTGAGAGAATCACTGTTCTTAGGAGCACATATTCTGGTCCAGCTAATCATGA GGTTTATTTAATTTGTATCCTTGCCAACAACTTTGCATTCCAGACTGTCTCGCAGGCAACAGGAATGGCGATGACT TTAACCGATCAAGGGTTTTACTTTGATATCACCTGAAAGAATAAATCAGTATTGGGATGGTCACTTGAAGCAAGAAC GTATCGTAGCAGAAGCAATTGATAAGGTGGTTCTAGGAGAAGATGCTCCTATTCAATTCGAGTGATAATGAATTAAT TCTCAAATGTGGAGGGACACCAAATGCACGGAATCTTATCGATATCGAGCCAGTCGCAACTTTCATAGAATTTGAA CAACTGATCTGCACAATGTTGACAACCCACTTGAAGGAAATAATTGATATAACAAGGTCTGGAACCCAGGATTATG AAAGTTATTACTCACTCCTTACAATTTAGGTCTTCTTGGTAAAATCAGTACGATAGTGAGATTATTAACAGAAAG GATTCTAAATCATACTATCAGGAATTGGTTGATCCTCCCACCTTCGCTCCGGATGATCGTGAAGCAGGACTTGGAA TTCGGCATATTCAGGATTACTTCCATCCTCAATTCTGATCGGTTCCTGAAGCTTTCTCCAAATAGGAAATACTTGA TTACACAATTAACTGCAGGCTACATTAGGAAATTGATTGAGGGGGATTGTAATATCGATCTAACCAGACCTATCCA AAAACAAATCTGGAAAGCATTAGGTTGTGTAGTCTATTGTCACGATCCAGTAGATCAAAGGGAATCAACAGAGTTT ATTGATATAAATATTAATGAAGAAATAGACCGCGGGATCGATGGCGAGGAAATCTAAATATATCAAGAATCAGAAT TAGTTTAAGAAAAAAGAAGAGGATTAATCTTGGTTTTCCCCTTGGT

[0158] The underlined sequence is the PIV5-CPI genomic nucleic acid sequence. The italicized and underlined sequences are the entire MOMP nucleic acid sequence. The double-underlined sequence is the AOS-modified NT-MOMPΔsp nucleic acid sequence.

[0159] 3. CVB vaccine sequence

[0160] a. pMHD181- CVB-MOMPΔsp-CD5sp

[0161] The pMHD181-CPI-MOMPΔsp-CD5sp nucleic acid sequence provided in this article, from 5' to 3', is as follows:

[0162] GTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGC GCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGC TGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCT GGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACT CTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGA AAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATG ACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTG ACACATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGT CAGCGGGTGTTGGCGGGTGTCGGGGCTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATATA TGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCTAATACGACTCACTATAGGGACCAAG GGGAAAATGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCTATGGCCTTCGTGACC GACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCGACACAAAAATGTCATCC GTGCTTAAAGCATATGAGCGATTCACGCTCACTCAAGAACTGCAAGATCAGAGTGAGGAAGGTACAATCCCACCTAC AACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGATCCCGGCTTCTTCTATTCT GCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACTCACAATGTTTTCGCTACCA TCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATCGAAAGGGTAGAGATCGATGG CTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCTCGTTCAGGTATGAGCCGTGGAGAGATCAATGCCTATGCTG CACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTCGTTGATTCCGAAGTCGAGGGAACTGCATGG GATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGGATAGTGACTTGCAAGTGCATGAC TGGCCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAAATACGTCAGCAAGGCAGGATCAACCCGAGAT ATCTCCTGCAACCGGAGGCTCGACGAATAATCCAGAATGTAATCCGGAAGGGAATGGTGGTCAGACATTTCCTCACC TTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTATGCTATGTAGGGGATGTTGGAAAGTA TATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAATATGCATTAGGAACTAGATGGCCCACACTTGCTT TAGCTGCATTTTCAGGAGAGCTAAACAAAGCTAAAGTCCCTCATGGCATTATACCAGACCCTTGGTGAGCAGGCCCGA TATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTGCTGCAGCAAACTACCCACTGCTATATAGCTATGCTAT GGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTCTCCAGATCATACATGAACAAGACATATTTCC AATTGGGAATGGAAACTGCAAGAAAACAACAGGGTGCAGTTGACATGAGGATGGCAGAAGATCTCGGTCTAACTCAA GCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACACAGCAAATCGAGGGGCAGACACCAGGGGAGGAGT CAACCCGTTCTCATCTGTCACTGGGACAACTCAGGTGCCCGCTGCAGCAACAGGTGACACACTCGAGAGTTACATGG CAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCATGATGATGAGATGCCACCATTGGAAGAGGAGGAA GAGGACGACACATCTGCAGGTCCACGCACTGGACCAACTCTTGAACAAGTGGCCTTGGACATCCAGAACGCAGCAGT TGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGGGTGATCTTGACATCTAGACAATTCAGATCCCAATCT AAAATTGACATACCTAATTGATTAGTTAGATGGAACTACAGTGGATTCCATAAGGTTCCTGCCTACCATCGGCTTTA AAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGGTGCCAACAGCGCAATCCACAATCTACAATGGA TCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATAAGCTCATAGAGACAGGCCTGAATACTGTAGAGTATTTTA CTTCCCAACAAGTCACAGGAACATCCTCTCTTGGAAAGAATACAATACCACCAGGGGTCACAGGACTACTAACCAAT GCTGCAGAGGCAAAGATCCAAGAGTCAACTAACCATCAGAAGGGCTCAGTTGGTGGGGGTGCAAAACCAAAGAAACC GCGACCAAAAATTGCCATTGTGCCAGCAGATGACAAAACAGTGCCCGGAAAGCCGATCCCAAACCCTCTATTAGGTC TGGACTCCACCCCGAGCACCCAAACTGTGCTTGATCTAAGTGGGAAAACATTACCATCAGGATCCTATAAGGGGGTT AAGCTTGCGAAATTTGGAAAAGAAAATCTGATGACACGGTTCATCGAGGAACCCAGAGAGAATCCTATCGCAACCAG TTTCCCCATCGATTTTAAGAGGGGCAGGGATACCGGCGGGTTCCATAGAAGGGAGTACTCAATCGGATGGGTGGGAG ATGAAGTCAAGGTCACTGAGTGGTGCAATCCATCCTGTTCTCCAATCACCGCTGCAGCAAGGCGATTTGAATGCACT TGTCACCAGTGTCCAGTCACTTGCTCTGAATGTGAACGAGATACTTAATACAGTGAGAAATTTGGACTCTCGGATGA ATCAACTGGAGACAAAAGTAGATCGCATTCTCTCATCTCAGTCTCTAATCCAGACCATCAAGAATGACATAGTTGGA CTTAAAGCAGGGATGGCTACTTTAGAAGGAATGATTACAACTGTGAAAATCATGGACCCGGGAGTTCCCAGTAATGT TACTGTGGAAGATGTACGCAAGACACTAAGTAACCATGCTGTTGTTGTGCCAGAATCATTCAATGATAGTTTCTTGA CTCAATCTGAAGATGTAATTTCACTTGATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAGAAGATTGTCAGGAAG GTTCCTCCTCAGAAGGATCTGACTGGATTGAAGATTACACTAGAGCAATTGGCAAAGGATTGCATCAGCAAACCGAA GATGAGGGAAGAGTATCTCCTCAAAATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCTCAAGAAAGCAATCA TCCGCAGTGCAATTTGATCAAGAAACACCCAATTACACTACACTGGTATGACACTGTACTAACCCTGAGGGTTTTAG AAAAAACGATTAACGATAAATAAGCCCGAACACTACACACTACCTGAGGCAGCCATGCCATCCATCAGCATTCCCGC AGACCCCACCAATCCACGTCAATCAATAAAAGCGTTCCCAATTGTGATCAACAGTGATGGGGGTGAGAAAGGCCGCT TGGTTAAACAACTACGCACAACCTACTTGAATGACCTAGATACTCATGAGCCACTGGTGACATTCATAAATACCTAT GGATTCATCTACGAACAGGATCGGGGGAATACCATTGTCGGAGAGGATCAACTTGGGAAGAAAAGAGAGGCTGTGAC CGCTGCAATGGTTACCCTTGGATGTGGGCCTAATCTACCATCATTAGGGAATGTCCTGGGACAACTGAGGGAATTCC AGGTCACTGTTAGGAAGACATCCAGCAAAGCGGAAGAGATGGTCTTTGAAATTGTTAAGTATCCGAGAATATTTCGG GGTCATACATTAATCCAGAAAGGACTAGTCTGTGTCTCCGCAGAAAAATTTGTTAAGTCACCAGGGAAAATACAATC TGGAATGGACTATCTCTTCATTCCGACATTTCTGTCAGTGACTTACTGTCCAGCTGCAATCAAATTTCAGGTACCTG GCCCCATGTTGAAAATGAGATCAAGATACACTCAGAGCTTACAACTTGAACTAATGATAAGAATCCTGTGTAAGCCC GATTCGCCACTTATGAAGGTCCATACCCCTGACAAGGAGGGAAGAGGATGTCTTGTATTCAGTATGGCTGCATGTATG CAACATCTTCAAATCAGGAAACAAGAATGGCAGTGAGTGGCAGGAATACTGGATGAGAAAGTGTGCTAACATGCAAC TTGAAGTGTCGATTGCAGATATGTGGGGACCAACTATCATAATTCATGCCAGAGGTCACATTCCCCAAGGTGCTAAG TTGTTTTTTGGAAAGGGTGGATGGAGCTGCCATCCACTTCACGAAGTTGTTCCAAGTGTCACTAAAACACTATGGTC CGTGGGCTGTGAGATTACAAAGGCGAAGGCAATAATACAAGAGAGTAGCATCTCTCTTCTCGTGGAGACTACTGACA TCATAAGTCCAAAAGTCAAAATTTCATCTAAGCATCGCCGCTTTGGGAAATCAAATTGGGGTCTGTTCAAGAAAACT AAATCACTGCCTAACCTGACGGAGCTGGAATGACTGACCTCTAATCGAGACTACACCGCCGCAAACTATAGGTGGGT GGTACCTCAGTGATTAATCTTGTAAGCACTGATCGTAGGCTACAACACACTAATATTATCCAGATTAGAGAGCTTAA TTAGCTCTGTATTAATAATAACACTACTATTCCAATAACTGGAATCACCAGCTTGATTTATCTCCAAAATGATTCAA AGAAAACAAATCATATTAAGACTATCCTAAGCACGAACCCATATCGTCCTTCAAATCATGGGTACTATAATTCAATT TCTGGTGGTCTCCTGTCTATTGGCAGGAGCAGGCAGCCTTGATCCAGCAGCCCTCATGCAAATCGGTGTCATTCCAA CAAATGTCCGGCAACTTATGTATTATACTGAAGCTTCATCAGCATTCATTGTTGTGAAGTTAATGCCTACAATTGAC TCGCCGATTAGTGGATGTAATATAACATCAATTTCAAGCTATAATGCAACAGTGACAAAACTCCTACAGCCGATCGG TGAGAATTTGGAGACGATTAGGAACCAGTTGATTCCAACTCGGAGGAGACGCCGGTTTGCAGGGGTGGTGATTGGAT TAGCTGCATTAGGAGTAGCTACTGCCGCACAGGTCACTGCCGCAGTGGCACTAGTAAAGGCAAATGAAAATGCTGCG GCTATACTCAATCTCAAAAATGCAATCCAAAAAACAAATGCAGCAGTTGCAGATGTGGTCCAGGCCACACAATCACT AGGAACGGCAGTTCAAGCAGTTCAAGATCACATAAACAGTGTGGTAAGTCCAGCAATTACAGCAGCCAATTGTAAGG CCCAAGATGCTATCATTGGCTCAATCCTCAATCTCTATTTGACCGAGTTGACAACCATCTTCCACAATCAAATTACA AACCCTGCATTGAGTCCCATTACAATTCAAGCTTTAAGGATCCTACTGGGGAGTACCTTGCCGACTGTGGTCGAAAA ATCTTTCAATACCCAGATAAGTGCAGCTGAGCTTCTCTCATCAGGGTTATTGACAGGCCAGATTGTGGGATTAGATT TGACCTATATGCAGATGGTCATAAAAATTGAGCTGCCAACTTTAACTGTACAACCTGCAACCCAGATCATAGATCTG GCCACCATTTCTGCATTCATTAACAATCAAGAAGTCATGGCCCAATTACCAACACGTGTTATGGTGACTGGCAGCTT GATCCAAGCCTATCCCGCATCGCAATGCACCATTACACCCAACACTGTGTACTGTAGGTATAATGATGCCCAAGTAC TCTCAGATGATACTATGGCTTGCCTCCAAGGTAACTTGACAAGATGCACCTTCTCTCCAGTGGTTGGGAGCTTTCTC ACTCGATTCGTGCTGTTCGATGGAATAGTTTATGCAAATTGCAGGTCGATGTTGTGCAAGTGCATGCAACCTGCTGC TGTGATCCTACAGCCGAGTTCATCCCCTGTAACTGTCATTGACATGTACAAATGTGTGAGTCTGCAGCTTGACAATC TCAGATTCACCATCACTCAATTGGCCAATGTAACCTACAATAGCACCATCAAGCTTGAATCATCCCAGATCTTGTCT ATTGATCCGTTGGATATATCCCAGAATCTAGCTGCGGTGAATAAGAGTCTAAGTGATGCACTACAACACTTAGCACA AAGTGACACATATCTTTCTGCAATCACATCAGCTACGACTACAAGTGTATTATCCATAATAGCAATCTGTCTTGGAT CTTAGGTTTAATATTAATAATCTTGCTCAGTGTAGTTGTGTGGAAGTTATTGACCATTGTCGTTGCTAATCGAAAT AGAATGGAGAATTTTGTTTATCATAAATAAGCATTCCACCACTCACGATCTGATCTCAGTGAGAAAAATCAACCTGC AACTCTTGGAACAAGATAAGACAGTCATCCATTAGTAATTTTTAAGAAAAAAACGATAGGACCGAACCTAGTATTGA AAGAACCGTCTCGGTCAATCTAGGTAATCGAGCTGATACCGTC

[0163]

[0164] ATGATCGACGGCATCCTGTGGGAGGGCTTCGGAGGCGATCCTTGTGATCCCTGCACCACATGGTGCGA CGCCATCAGCCTTAGACTGGGCTACTACGGCGACTTCGTGTTTGATAGAGTGCTGAAAACCGATGTCAACAAGCAG TTTGAGATGGGCGCCGCTCCAACAGGCGACGCCGACCTGACCACCGCCCCTACACCAGCCAGCCGGGAAAACCCTG CCTACGGCAAGCACATGCAGGACGCCGAGATGTTTACAAACGCCGCTTATATGGCCCTGAATATCTGGGACCGGTT CGACGTGTTCTGCACCCTGGGCGCTACATCTGGATACCTGAAGGGCAACAGCGCCGCTTTCAACCTGGTGGGCCTG TTCGGCAGAGATGAGACGGCAGTTGCTGCTGATGACATCCCTAACGTGAGCCTGAGCCAGGCCGTTGTGGAACTGT ACACCGACACCGCCTTTGCCTGGAGCGTGGGAGCCAGAGCCGCCCTGTGGGAGTGCGGTTGTGCCACACTGGGCGC CAGCTTCCAGTACGCCCAGAGCAAGCCCAAAGTGGAAGAGCTGAATGTGCTGTGCAACGCCGCAGAGTTCACCATC AACAAGCCTAAGGGCTATGTGGGCCAGGAGTTCCCCCTGAACATCAAGGCCGGCACCGTGTCCGCCACCGACACCA AGGACGCCAGCATCGATTACCACGAGTGGCAGGCCTCTCTGGCCCTCAGCTACAGACTGAACATGTTCACACCTTA CATCGGCGTGAAGTGGTCCCGCGCCAGCTTCGACGCTGATACAATCAGAATCGCCCAACCTAAGCTGGAAACCTCT ATTCTGAAGATGACTACCTGGAACCCCACCATCAGCGGCAGCGGCATCGACGTGGACACCAAGATCACCGACACCC TGCAGATCGTGTCCCTGCAACTGAACAAGATGAAAAGCAGAAAGAGCTGCGGCCTGGCTATTGGAACAACCATCGT GGACGCCGATAAGTACGCCGTGACCGTGGAAACAAGACTGATCGACGAGCGGGCCGCCCACGTGAACGCCCAGTTC CGGTTCTGAGTCGACCACCTGCTATAGGCTATCCACTGCATCATCTCTCCTGCCATACTTCCTACTCACATCATAT CTATTTTAAAGAAAAAATAGGCCCGAACACTAATCGTGCCGGCAGTGCCACTGCACACACAACACTACACATACAA TACACTACAATGGTTGCAGAAGATGCCCCTGTTAGGGCCACTTGCCGAGTATTATTTCGAACAACAACTTTAATCT TTCTATGCACACTACTAGCATTAAGCATCTCTATCCTTTATGAGAGTTTAATAACCCAAAAGCAAATCATGAGCCA AGCAGGCTCAACTGGATCTAATTCTGGATTAGGAAGTATCACTGATCTTCTTAATAATATTCTCTCTGTCGCAAAT CAGATTATATATAACTCTGCAGTCGCTCTACCTCTACAATTGGACACTCTTGAATCAACACTCCTTACAGCCATTA AGTCTCTTCAAACCAGTGACAAGCTAGAACAGAACTGCTCGTGGAGTGCTGCACTGATTAATGATAATAGATACAT TAATGGCATCAATCAGTTCTATTTTTCAATTGCTGAGGGTCGCAATCTGACACTTGGCCCACTTCTTAATATGCCT AGTTTCATTCCAACTGCCACGACACCAGAGGGCTGCACCAGGATCCCATCATTCTCGCTCACTAAGACACACTGGT GTTATACACACAATGTTATCCTGAATGGATGCCAGGATCATGTATCCTCAAATCAATTTGTTTCTATGGGAATCAT TGAACCCACTTCTGCCGGGTTTCCATTCTTTCGAACCCTAAAGACTCTATATCTCAGCGATGGGGTCAATCGTAAG AGCTGCTCTATCAGTACAGTTCCGGGGGGTTGTATGATGTACTGTTTTGTTTCTACTCAACCAGAGAGGGATGACT ACTTTTCTGCCGCTCCTCCAGAACAACGAATTATTATAATGTACTATAATGATACAATCGTGGAGCGCATAATTAA TCCACCCGGGGTACTAGATGTATGGGCAACATTGAACCCAGGAACAGGAAGCGGGGTATATTATTTAGGTTGGGTG CTCTTTCCAATATATGGCGGCGTGATTAAAGGTACGAGTTTATGGAATAATCAAGCAAATAAATACTTTATCCCCC AGATGGTTGCTGCTCTCTGCTCACAAAACCAGGCAACTCAAGTCCAAAATGCTAAGTCATCATACTATAGCAGCTG GTTTGGCAATCGAATGATTCAGTCTGGGATCCTGGCATGTCCTCTTCGACAGGATCTAACCAATGAGTGTTTAGTT CTGCCCTTTTCTAATGATCAGGTGCTTATGGGTGCTGAAGGGAGATTATACATGTATGGTGACTCGGTGTATTACT ATCAAAGAAGCAATAGTTGGTGGCCTATGACCATGCTGTATAAGGTAACCATAACATTCACTAATGGTCAGCCATC TGCTATATCAGCTCAGAATGTGCCCACACAGCAGGTCCCTAGACCTGGGACAGGAGACTGCTCTGCAACCAATAGA TGTCCCGGTTTTTGCTTGACAGGAGTGTATGCCGATGCCTGGTTACTGACCAACCCTTCGTCTACCAGTACATTTG GATCAGAAGCAACCTTCACTGGTTCTTATCTCAACACAGCAACTCAGCGTATCAATCCGACGATGTATATCGCGAA CAACACACAGATCATAAGCTCACAGCAATTTGGATCAAGCGGTCAAGAAGCAGCATATGGCCACACAACTTGTTTT AGGGACACAGGCTCTGTTATGGTATACTGTATCTATATTATTGAATTGTCCTCATCTCTCTTAGGACAATTTCAGA TTGTCCCATTTATCCGTCAGGTGACACTATCCTAAAGGCAGAAGCCTTCAGGTCTGACCCAGCCAATCAAAGCATT ATACCAGACCATGGAATGCATACCAAACATTATTGACACTAATGACACACAAAATTGGTTTTAAGAAAAACCAAGA GAACAATAGGCCAGAATGGCTGGGTCTCGGGAGATATTACTCCCTGAAGTCCATCTCAATTCACCAATTGTAAAGC ATAAGCTATACTATTACATTCTACTTGGAAACCTCCCAAATGAGATCGACCTTGACGATTTAGGTCCATTACATAA TCAAAATTGGAATCAGATAGCACATGAAGAGTCTAACTTAGCTCAACGCTTGGTAAATGTAAGAAATTTTCTAATT ACCCACATCCCTGATCTTAGAAAGGGCCATTGGCAAGAGTATGTCAATGTAATACTGTGGCCGCGAATTCTTCCCT TGATCCCGGATTTTAAAATCAATGACCAATTGCCTCTGCTCAAAAATTGGGACAAGTTAGTTAAAGAATCATGTTC AGTAATCAATGCAGGTACTTCCCAGTGCATTCAGAATCTCAGCTATGGACTGACAGGTCGTGGGAACCTCTTTACA CGATCACGTGAACTCTCTGGTGACCGCAGGGATATTGATCTTAAGACAGTTGTGGCAGCATGGCATGACTCAGACT GGAAAAGAATAAGTGATTTTTGGATTATGATCAAATTCCAGATGAGACAATTAATTGTTAGGCAAACAGATCATAA TGATTCTGATTTAATCACGTATATCGAAAATAGAGAAGGCATAATCATCATAACCCCTGAACTGGTAGCATTATTT AACACTGAGAATCATACACTAACATACATGACCTTTGAAATTGTACTGATGGTTTCAGATATGTACGAAGGTCGTC ACAACATTTTATCACTATGCACAGTTAGCACTTACCTGAATCCTCTGAAGAAAAGAATAACATATTTATTGAGCCT TGTAGATAACTTAGCTTTTCAGATAGGTGATGCTGTATATAACATAATTGCTTTGCTAGAATCCTTTGTATATGCA CAGTTGCAAATGTCAGATCCCATCCCAGAACTCAGAGGACAATTCCATGCATTCGTATGTTCTGAGATTCTTGATG CACTAAGAGGAACTAATAGTTTCACCCAGGATGAATTAAGAACTGTGACAACTAATTTGATATCCCATTCCAAGA TCTGACCCCAGATCTTACGGCTGAATTGCTCTGTATAATGAGGCTTTGGGGACACCCCATGCTCACTGCCAGTCAA GCTGCAGGAAAGGTACGCGAGTTCTATGTGTGCTGGAAAAGTATTAGACTTTCCCCACCATTATGAAAACACTAGCCT TTTTCCATACTATTCTGATCAATGGATACAGGAGGAAGCATCATGGAGTATGGCCACCCTTAAACTTACCGGGTAA TGCTTCAAAGGGTCTCACGGAACTTATGAATGACAATACTGAAATAAGCTATGAATTCACACTTAAGCATTGGAAG GAAGTCTCTCTTATAAAATTCAAGAAATGTTTTGATGCAGACGCAGGTGAGGAACTCAGTATATTTATGAAAGATA AGGCAATTAGTGCCCCAAAACAAGACTGGATGAGTGTGTTTAGAAGAAGCCTAATCAAACAGCGCCATCAGCATCA TCAGGTCCCCCTACCAAATCCATTCAATCGACGGCTGTTGCTAAACTTTTCTCCGGAGATGACAAATTCGACCCGAAT GTGGAGCTACAGTATGTAACATCAGGTGAGTATCTACATGATGACACGTTTTGTGCATCATATTCACTAAAAGAGA AGGAAATTAAACCTGATGGTCGAATTTTTGCAAAGTTGACTAAGAGAATGAGATCATGTCAAGTTATAGCAGAATC TCTTTAGCGAACCATGCTGGGAAGTTAATGAAAGAGAATGGTGTTGTGATGAATCAGCTATCATTAACAAAATCA CTATTAACAATGAGTCAGATTGGAATAATATCCGAGAAAGCTAGAAAGTCAACTCGAGATAACATAAATCAACCTG GTTTCCAGAATATCCAGAGAAATAAATCACATCACTCCAAGCAAGTCAATCAGCGAGATCCAAGTGATGACTTTGA ATTGGCAGCATCTTTTTTAACTACTGATCTCAAAATATTGTTTACAATGGAGGTACCAGACAATTATCCCATTT GCTCAATCATTAAACAGAATGTATGGTTATTCCTCATCTCTTTGAGTGGATTCACTTACGGCTAATGCGTAGTACAC TTTACGTGGGGGATCCCTTCAACCCACCAGCAGATACCAGTCAATTTGATCTAGATAAAGTAATTAATGGAGATAT CTTCATTGTATCACCCAGAGGTGGAATTGAAGGGCTGTGTCAAAAGGCTTGGACAATGATATCTATCGCTGTGATA ATTCTATTCTGCCACAGAGTCTGGCACACGAGTAATGAGTATGGTGCAGGGAGATAATCAAGCAATTGCTGTCACCA CACGAGTACCAAGGAGCCTGCCGACTCTTGAGAAAAAGACTATTGCTTTAGATCTTGTAATCTATTCTTTGAGAG GTTAAAATGTAATAATTTTGGATTAGGTCACCATTTGAAAGAACAAGAGACTATCATTAGTTCTCACTTCTTTGTT TATAGCAAGAGAATATTCTATCAGGGGAGGATTCTAACGCAAGCCTTAAAAAATGCTAGTAAGCTCTGCTTGACAG CTGAGTCCTAGGAGAATGCACCCAATCATCATGTTCTAATCTTGCAACTACTGTCATGAGGTTAACTGAGAATGG TGTTGAAAAAGATATCTGTTTCTACTTGAATATCTATATGACCATCAAACAGCTCTCCTATGATATCATCTTCCCT CAAGTGTCAATTCCTGGAGATCAGATCACATTAGAATACATAAATAATCCACACCTGGTATCACGATTGGCTCTTT TGCCATCCCAGTTAGGAGGTCTAAACTACCTGTCATGCAGTAGGCTGTTCAATCGAAACATAGGCGACCCGGTGGT TTCCGCAGTTGCAGATCTTAAGAGATTAATTAAATCAGGATGTATGGATTACTGGATCCTTTATAACTTATTAGGG AGAAAACCGGGAAACGGCTCATGGGCTACTTTAGCAGCTGACCCGTACTCAATCAATATAGAGTATCAATACCCTC CAACTACAGCTCTTAAGAGGCACACCCAACAAGCTCTGATGGAACTCAGTACGAATCCAATGTTACGTGGCATATT CTCTGACAATGCACAGGCAGAAGAAAATAACCTTGCTAGGTTTCTCCTGGATAGGGAGGTGATCTTTCCGCGTGTA GCTCACATCATCATTGAGCAAACCAGTGTCGGGAGGAGAAAACAGATTCAAGGATATTTGGATTCAACTAGATCGA TAATGAGGAAATCACTAGAAATTAAGCCCTTATCCAATAGGAAGCTTAATGAAATACTGGATTACAACATCAATTA CCTAGCTTACAATTTGGCATTACTCAAGAATGCTATTGAACCTCCGACTTATTTGAAGGCAATGACACTTGAAACA TGTAGCATCGACATTGCAAGGAACCTCCGGAAGCTCTCCTGGGCCCCACTCTTGGGTGGGAGAAATCTTGAAGGAT TAGAGACGCCAGATCCCATTGAAATTACTGCAGGAGCATTAATTGTTGGATCGGGCTACTGTGAACAGTGTGCTGC AGGAGACAATCGATTCACATGGTTTTTCTTGCCATCTGGTATCGAGATAGGAGGGGATCCCCGTGATAATCCTCCT ATCCGTGTACCGTACATTGGCTCCAGGACTGATGAGAGGAGGGTAGCCTCAATGGCATACATCAGGGGTGCCTCGA GTAGCTTAAAAGCAGTTCTTAGACTGGCGGGAGTGTACATCTGGGCATTCGGAGATACTCTGGAGAATTGGATAGA TGCACTGGATTTGTCTCACACTAGAGTTAACATCACACTTGAACAGCTGCAATCCCTCACCCCACTTCCAACCTCT GCCAATCTAACCCATCGGTTGGATGATGGCACAACTACCCTAAAGTTTACTCCTGCGAGCTCTTACACCTTTTCAA GTTTCACTCATATATCAAATGATGAGCAATACCTGACAATTAATGACAAAACTGCAGATTCAAATATAATCTACCA ACAGTTAATGATCACTGGACTCGGAATCTTAGAAACATGGAATAATCCCCCAATCAATAGAACATTCGAAGAATCT ACCCTACATTTGCACACTGGTGCATCATGTTGTGTCCGACCTGTGGACTCCTGCATTCTCTCAGAAGCATTAACAG TCAAGCCACATATTACAGTACCGTACAGCAATAAATTTGTATTTGATGAGGACCCGCTATCTGAATATGAAACTGC AAAACTGGAATCGTTATTCATTCCAAGCCCAATTAGGCAACATTGATGCTGTAGATATGACAGGTAAATTAACATTA TTGTCCCAATTCACTGCAAGGCAGATTATCAATGCAATCACTGGACTCGATGAGTCTGTCTCTCTTACTAATGATG CCATTGTTGCATCAGACTATGTCTCCAATTGGATTAGTGAATGCATGTATACCAAAATTAGATGAATTATTTATGTA TTGTGGGTGGGAACTACTATTGGAACTATCCTATCAAATGTATTATCTGAGGGTAGTTGGGTGGAGTAATATAGTG GATTATTCTTACATGATCTTGAGAAGAATCCCGGGTGCAGCATTAAACAATCTGGCATCTACATTAAGTCATCCAA AACTTTTCCGACGAGCTATCAACCTAGATATAGTTGCCCCCTTAAATGCTCCTCATTTTGCATCTCTGGACTACAT CAAGATGAGTGTGGATGCAATACTCTGGGGCTGTAAAAGAGTCATCAATGTCTCTCCAATGGAGGGGACTTAGAA TTAGTTGTGACATCTGAAGATAGCCTTATTCTCAGTGACCGATCCATGAATCTCATTGCAAGGAAATTAACTTTAT TATCACTGATTCACCATAATGGTTTGGAACTACCAAAGATTAAGGGGTTCTCTCCTGATGAGAAGTGTTTCGCTTT GACAGAATTTTTGAGGAAAGTGGTGAACTCAGGGTTGAGTTCAATAGAGAACCTATCAAATTTTATGTACAATGTG GAGAACCCACGGCTTGCAGCATTCGCCAGCAACAATTACTACCTGACCAGAAAATTATTGAATTCAATACGAGATA CTGAGTCGGGTCAAGTAGCAGTCACCTCATATTATGAATCATTAGAATATATTGATAGTCTTAAGCTAACCCCACA TGTGCCTGGCACCTCATGCATTGAGGATGATAGTCTATGTACAAATGATTACATAATCTGGATCATAGAGTCTAAT GCAAACTTGGAGAAGTATCCAATTCCAAATAGCCCTGAGGATGATTCCAATTTCCATAACTTTAAGTTGAATGCTC CATCGCACCATACCTTACGCCCATTAGGGTTGTCATCAACTGCTTGGTATAAGGGTATAAGCTGCTGCAGGTACCT TGAGCGATTAAAGCTACCACAAGGTGATCATTTATATATTGCAGAAGGTAGTGGTGCCAGTATGACAATCATAGAA TACCTATTCCCAGGAAGAAAGATATATTACAATTCTTTATTTAGTAGTGGTGACAATCCCCCACAAAGAAATTATG CACCAATGCCTACTCAGTTCATTGAGAGTGTCCCATACAAGCTCTGGCAAGCACACACAGATCAATATCCCGAGAT TTTTGAGGACTTCATCCCTCTATGGAACGGAAACGCCGCCATGACTGACATAGGAATGACAGCTTGTGTAGAATTC ATCATCAATCGAGTCGGCCCAAGGACTTGCAGTTTAGTACATGTAGATTTGGAATCAAGTGCAAGCTTAAATCAAC AATGCCTGTCAAAGCCGATAATTAATGCTATCATCACTGCTACAACTGTTTTGTGCCCTCATGGGGTGCTTATTCT GAAATATAGTTGGTTGCCATTTACTAGATTTAGTACTTTGATCACTTTCTTATGGTGCTACTTTGAGAGAATCACT GTTCTTAGGAGCACATATTCTGATCCAGCTAATCATGAGGTTTATTTAATTTGTATCCTTGCCAACAACTTTGCAT TCCAGACTGTCTCGCAGGCAACAGGAATGGCGATGACTTTAACTGATCAAGGGTTTACTTTGATATCACCTGAAAG AATAAATCAGTATTGGGATGGTCACTTGAAGCAAGAACGTATCGTAGCAGAAGCAATTGATAAGGTGGTTCTAGGA GAAAATGCTCTATTTAATTCGAGTGATAATGAATTAATTCTCAAATGTGGAGGGACACCAAATGCACGGAATCTCA TCGATATCGAGCCAGTCGCAACTTTCATAGAATTTGAACAATTGATCTGCACAATGTTGACAACCCACTTGAAGGA AATAATTGATATAACAAGGTCTGGAACCCAGGATTATGAAAGTTTATTACTCACTCCTTACAATTTAGGTCTTCTT GGTAAAATCAGTACGATAGTGAGATTATTAACAGAAAGGATTCTAAATCATACTATCAGGAATTGGTTGATCCTCC CACCTTCGCTCCGGATGATCGTGAAGCAGGACTTGGAATTCGGCATATTCAGGATTACTTCCATCCTCAATTCTGA TCGGTTCCTGAAGCTTTCTCCAAATAGGAAATACTTGATTGCACATTAACTGCAGGCTACATTAGGAAATTGATT GAGGGGGATTGCAATATCGATCTAACCAGACCTATCCAAGCAAATCTGGAAAGCATTAGGTTGTGTAGTCTATT GTCACGATCCAATGGATCAAAGGGAGTCAACAGAGTTTATTGATATAAATATTAATGAAGAAAATAGACCGCGGGAT CGATGGCGAGGAAATCTAAACATATCAAGAATCAGAATTAGTTTAAGAAAAAAGAAGAGGATTAATCTTGGTTTTC CCCTTGGT

[0165] The underlined sequence is the PIV5-CVB genomic nucleic acid sequence. The italicized and underlined sequences are the entire MOMP nucleic acid sequence. The double-underlined sequence is the AOS-modified MOMPΔsp-CD5sp nucleic acid sequence.

[0166] b. pMHD182- CVB-NT-MOMPΔsp

[0167] The pMHD182-CPI-NT-MOMPΔsp nucleic acid sequence provided in this article, from 5' to 3', is as follows:

[0168] ACCAAGGGGAAAATGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCTA TGGCCTTCGTGACCGACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCGACA CAAAAATGTCATCCGTGCTTAAAGCATATGAGCGATTCACGCTCACTCAAGAACTGCAAGATCAGAGTGAGGAAGGT ACAATCCCACCTACAACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGATCCCG GCTTCTTCTATTCTGCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACTCACAA TGTTTTCGCTACCATCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATCGAAAGG GTAGAGATCGATGGCTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCTCGTTCAGGTATGAGCCGTGGAGAGAT CAATGCCTATGCTGCACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTCGTTGATTCCGAAGTCG AGGGAACTGCATGGGATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGGATAGTGACT TGCAAGTGCATGACTGCGCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAATATCGTCAGCAAGGCAG GATCAACCCGAGATATCTCCTGCAACCGGAGGCTCGACGAATAATCCAGAATGTAATCCGGAAGGGAATGGTGGTCA GACATTTCCTCACCTTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTATGCTATGGTAGGG GATGTTGGAAAGTATATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAATATGCATTAGGAACTAGATG GCCCACACTTGCTTTAGCTGCATTTTCAGGAGAGCTAACAAAGCTAAAGTCCCTCATGGCATTATACCAGACCCTTG GTGAGCAGGCCCGATATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTTGCTGCAGCAAACTACCCACTGCTA TATAGCTATGCTATGGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTCTCCAGATCATACATGAA CAAGACATATTTCCAATTGGGAATGGAAACTGCAAGAAAACAACAGGGTGCAGTTGACATGAGGATGGCAGAAGATC TCGGTCTAACTCAAGCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACAGCAAATCGAGGGGCAGAC ACCAGGGGAGGAGTCAACCCGTTCTCATCTGTCACTGGGACAACTCAGGTGCCCGCTGCAGCAACAGGTGACACACT CGAGAGTTACATGGCAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCATGATGATGAGATGCCACCAT TGGAAGAGGAGGAAGAGGACGACACATCTGCAGGTCCACGCACTGGACCAACTCTTGAACAAGTGGCCTTGGACATC CAGAACGCAGCAGTTGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGGGTGATCTTGACATCTAGACAAT TCAGATCCCAATCTAAAATTGACATACCTAATTGATTAGTTAGATGGAACTACAGTGGATTCCATAAGGTTCCTGCC TACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGGTGCCAACAGCGCAATCCA CAATCTACAATGGATCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATAAGCTCATAGAGACAGGCCTGAATAC TGTAGAGTATTTTACTTCCCAACAAGTCACAGGAACATCCTCTCTTGGAAAGAATACAATACCACCAGGGGTCACAG GACTACTAACCAATGCTGCAGAGGCAAAGATCCAAGAGTCAACTAACCATCAGAAGGGCTCAGTTGGTGGGGGTGCA AAACCAAAGAAACCGCGACCAAAAATTGCCATTGTGCCAGCAGATGACAAAACAGTGCCCGGAAAGCCGATCCCAAA CCCTCTATTAGGTCTGGACTCCACCCCGAGCACCCAAACTGTGCTTGATCTAAGTGGGAAAACATTACCATCAGGAT CCTATAAGGGGGTTAAGCTTGCGAAATTTGGAAAAGAAAATCTGATGACACGGTTCATCGAGGAACCCAGAGAGAAT CCTATCGCAACCAGTTTCCCCATCGATTTTAAGAGGGGCAGGGATACCGGCGGGTTCCATAGAAGGGAGTACTCAAT CGGATGGGTGGGAGATGAAGTCAAGGTCACTGAGTGGTGCAATCCATCCTGTTCTCCAATCACCGCTGCAGCAAGGC GATTTGAATGCACTTGTCACCAGTGTCCAGTCACTTGCTCTGAATGTGAACGAGATACTTAATACAGTGAGAAATTT GGACTCTCGGATGAATCAACTGGAGACAAAAGTAGATCGCATTCTCTCATCTCAGTCTCTAATCCAGACCATCAAGA ATGACATAGTTGGACTTAAAGCAGGGATGGCTACTTTAGAAGGAATGATTACAACTGTGAAAATCATGGACCCGGGA GTTCCCAGTAATGTTACTGTGGAAGATGTACGCAAGACACTAAGTAACCATGCTGTTGTTGTGCCAGAATCATTCAA TGATAGTTTCTTGACTCAATCTGAAGATGTAATTTCACTTGATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAGA AGATTGTCAGGAAGGTTCCTCCTCAGAAGGATCTGACTGGATTGAAGATTACACTAGAGCAATTGGCAAAGGATTGC ATCAGCAAACCGAAGATGAGGGAAGAGTATCTCCTCAAAATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCT CAAGAAAGCAATCATCCGCAGTGCAATTTGATCAAGAAACACCCAATTACACTACACTGGTATGACACTGTACTAAC CCTGAGGGTTTTAGAAAAAACGATTAACGATAAATAAGCCCGAACACTACACACTACCTGAGGCAGCCATGCCATCC ATCAGCATTCCCGCAGACCCCACCAATCCACGTCAATCAATAAAAGCGTTCCCAATTGTGATCAACAGTGATGGGGG TGAGAAAGGCCGCTTGGTTAAACAACTACGCACAACCTACTTGAATGACCTAGATACTCATGAGCCACTGGTGACAT TCATAAATACCTATGGATTCATCTACGAACAGGATCGGGGGAATACCATTGTCGGAGAGGATCAACTTGGGAAGAAA AGAGAGGCTGTGACCGCTGCAATGGTTACCCTTGGATGTGGGCCTAATCTACCATCATTAGGGAATGTCCTGGGACA ACTGAGGGAATTCCAGGTCACTGTTAGGAAGACATCCAGCAAAGCGGAAGAGATGGTCTTTGAAATTGTTAAGTATC CGAGAATATTTCGGGGTCATACATTAATCCAGAAAGGACTAGTCTGTGTCTCCGCAGAAAAATTTGTTAAGTCACCA GGGAAAATACAATCTGGAATGGACTATCTCTTCATTCCGACATTTCTGTCAGTGACTTACTGTCCAGCTGCAATCAA ATTTCAGGTACCTGGCCCCATGTTGAAAATGAGATCAAGATACACTCAGAGCTTACAACTTGAACTAATGATAAGAA TCCTGTGTAAGCCCGATTCGCCACTTATGAAGGTCCATACCCCTGACAAGGAGGGAAGAGGATGTCTTGTATCAGTA TGGCTGCATGTATGCAACATCTTCAAATCAGGAAACAAGAATGGCAGTGAGTGGCAGGAATACTGGATGAGAAAGTG TGCTAACATGCAACTTGAAGTGTCGATTGCAGATATGTGGGGACCAACTATCATAATTCATGCCAGAGGTCACATTC CCAAAAGTGCTAAGTTGTTTTTTGGAAAGGGTGGATGGAGCTGCCATCCACTTCACGAAGTTGTTCCAAGTGTCACT AAAACACTATGGTCCGTGGGCTGTGAGATTACAAAGGCGAAGGCAATAATACAAGAGAGTAGCATCTCTCTTCTCGT GGAGACTACTGACATCATAAGTCCAAAAGTCAAAATTTCATCTAAGCATCGCCGCTTTGGGAAATCAAATTGGGGTC TGTTCAAGAAAACTAAATCACTGCCTAACCTGACGGAGCTGGAATGACTGACCTCTAATCGAGACTACACCGCCGCA AACTATAGGTGGGTGGTACCTCAGTGATTAATCTTGTAAGCACTGATCGTAGGCTACAACACACTAATATTATCCAG ATTAGAGAGCTTAATTAGCTCTGTATTAATAATAACACTACTATTCCAATAACTGGAATCACCAGCTTGATTTATCT CCAAAATGATTCAAAGAAAACAAATCATATTAAGACTATCCTAAGCACGAACCCATATCGTCCTTCAAATCATGGGT ACTATAATTCAATTTCTGGTGGTCTCCTGTCTATTGGCAGGAGCAGGCAGCCTTGATCCAGCAGCCCTCATGCAAAT CGGTGTCATTCCAACAAATGTCCGGCAACTTATGTATTATACTGAAGCTTCATCAGCATTCATTGTTGTGAAGTTAA TGCCTACAATTGACTCGCCGATTAGTGGATGTAATATAACATCAATTTCAAGCTATAATGCAACAGTGACAAAACTC CTACAGCCGATCGGTGAGAATTTGGAGACGATTAGGAACCAGTTGATTCCAACTCGGAGGAGACGCCGGTTTGCAGG GGTGGTGATTGGATTAGCTGCATTAGGAGTAGCTACTGCCGCACAGGTCACTGCCGCAGTGGCACTAGTAAAGGCAA ATGAAAATGCTGCGGCTATACTCAATCTCAAAAATGCAATCCAAAAAACAAATGCAGCAGTTGCAGATGTGGTCCAG GCCACACAATCACTAGGAACGGCAGTTCAAGCAGTTCAAGATCACATAAACAGTGTGGTAAGTCCAGCAATTACAGC AGCCAATTGTAAGGCCCAAGATGCTATCATTGGCTCAATCCTCAATCTCTATTTGACCGAGTTGACAACCATCTTCC ACAATCAAATTACAAACCCTGCATTGAGTCCCATTACAATTCAAGCTTTAAGGATCCTACTGGGGAGTACCTTGCCG ACTGTGGTCGAAAAATCTTTCAATACCCAGATAAGTGCAGCTGAGCTTCTCTCATCAGGGTTATTGACAGGCCAGAT TGTGGGATTAGATTTGACCTATATGCAGATGGTCATAAAAATTGAGCTGCCAACTTTAACTGTACAACCTGCAACCC AGATCATAGATCTGGCCACCATTTCTGCATTCATTAACAATCAAGAAGTCATGGCCCAATTACCAACACGTGTTATG GTGACTGGCAGCTTGATCCAAGCCTATCCCGCATCGCAATGCACCATTACACCCAACACTGTGTACTGTAGGTATAA TGATGCCCAAGTACTCTCAGATGATACTATGGCTTGCCTCCAAGGTAACTTGACAAGATGCACCTTCTCTCCAGTGG TTGGGAGCTTTCTCACTCGATTCGTGCTGTTCGATGGAATAGTTTATGCAAATTGCAGGTCGATGTTGTGCAAGTGC ATGCAACCTGCTGCTGTGATCCTACAGCCGAGTTCATCCCCTGTAACTGTCATTGACATGTACAAATGTGTGAGTCT GCAGCTTGACAATCTCAGATTCACCATCACTCAATTGGCCAATGTAACCTACAATAGCACCATCAAGCTTGAATCAT CCCAGATCTTGTCTATTGATCCGTTGGATATATCCCAGAATCTAGCTGCGGTGAATAAGAGTCTAAGTGATGCACTA CAACACTTAGCACAAAGTGACACATATCTTTCTGCAATCACATCAGCTACGACTACAAGTGTATTATCCATAATAGC AATCTGTCTTGGATCGTTAGGTTTAATATTAATAATCTTGCTCAGTGTAGTTGTGTGGAAGTTATTGACCATTGTCG TTGCTAATCGAAATAGAATGGAGAATTTTGTTTATCATAAATAAGCATTCCACCACTCACGATCTGATCTCAGTGAG AAAAATCAACCTGCAACTCTTGGAACAAGATAAGACAGTCATCCATTAGTAATTTTTAAGAAAAAAACGATAGGACC GAACCTAGTATTGAAAGAACCGTCTCGGTCAATCTAGGTAATCGAGCTGATACCGTCTCGGAAAGCTCAAGCGGCC

[0169]

[0170] CTGATGATCGACGGCATCCTGTGGGAGGGCTTCGGAGGCGATCCTTGTGATCCCTGCACCACATGGTG CGACGCCATCAGCCTTAGACTGGGCTACTACGGCGACTTCGTGTTTGATAGAGTGCTGAAAACCGATGTCAACAAG CAGTTTGAGATGGGCGCCGCTCCAACAGGCGACGCCGACCTGACCACCGCCCCTACACCAGCCAGCCGGGAAAACC CTGCCTACGGCAAGCACATGCAGGACGCCGAGATGTTTACAAACGCCGCTTATATGGCCCTGAATATCTGGGACCG GTTCGACGTGTTCTGCACCCTGGGCGCTACATCTGGATACCTGAAGGGCAACAGCGCCGCTTTCAACCTGGTGGGC CTGTTCGGCAGAGATGAGACGGCAGTTGCTGCTGATGACATCCCTAACGTGAGCCTGAGCCAGGCCGTTGTGGAAC TGTACACCGACACCGCCTTTGCCTGGAGCGTGGGAGCCAGAGCCGCCCTGTGGGAGTGCGGTTGTGCCACACTGGG CGCCAGCTTCCAGTACGCCCAGAGCAAGCCCAAAGTGGAAGAGCTGAATGTGCTGTGCAACGCCGCAGAGTTCACC ATCAACAAGCCTAAGGGCTATGTGGGCCAGGAGTTCCCCCTGAACATCAAGGCCGGCACCGTGTCCGCCACCGACA CCAAGGACGCCAGCATCGATTACCACGAGTGGCAGGCCTCTCTGGCCCTCAGCTACAGACTGAACATGTTCACACC TTACATCGGCGTGAAGTGGTCCCGCGCCAGCTTCGACGCTGATACAATCAGAATCGCCCAACCTAAGCTGGAAACC TCTATTCTGAAGATGACTACCTGGAACCCCACCATCAGCGGCAGCGGCATCGACGTGGACACCAAGATCACCGACA CCCTGCAGATCGTGTCCCTGCAACTGAACAAGATGAAAAGCAGAAAGAGCTGCGGCCTGGCTATTGGAACAACCAT CGTGGACGCCGATAAGTACGCCGTGACCGTGGAAACAAGACTGATCGACGAGCGGGCCGCCCACGTGAACGCCCAG TTCCGGTTCTGATGAGTCGACCACCTGCTATAGGCTATCCACTGCATCATCTCTCCTGCCATACTTCCTACTCACA TCATATCTATTTTAAAGAAAAAATAGGCCCGAACACTAATCGTGCCGGCAGTGCCACTGCACACACAACACTACAC ATACAATACACTACAATGGTTGCAGAAGATGCCCCTGTTAGGGCCACTTGCCGAGTATTATTTCGAACAACAACTT TAATCTTTCTATGCACACTACTAGCATTAAGCATCTCTATCCTTTGAGAGTTTAATAACCCAAAGCAAATCAT GAGCCAAGCAGGCTCAACTGGATCTAATTCTGGATTAGGAAGTATCACTGATCTTCTTAATAATATTCTCTCTGTC GCAAATCAGATTATATAACTCTGCAGTCGCTCTACCTCTACAATTGGACACTCTTTGAATCAACACTCCTTACAG CCATTAAGTCTCTTCAAACCAGTGACAAGCTAGAACAGAACTGCTCGTGGAGTGCTGCACTGATTAATGATAATAG ATACATTAATGGCATCAATCAGTTCTATTTTTCAATTGCTGAGGGTCGCAATCTGACACTTGGCCCACTTCTTAAT ATGCCTAGTTTCATTCCAACTGCCACGACACCAGAGGGCTGCACCAGGATCCCATTCTCGCTCACTAAGACAC ACTGGTGTTATACACACAATGTTATCCTGAATGGATGCCAGGATCATGTATCCTCAAATCAATTTGTTTCTATGGG AATCATTGAACCCACTTCTGCCGGGTTTCCATTCTTTCGAACCCTAAAGACTCTATATCTCAGCGATGGGGTCAAT CGTAAGAGCTGCTCTATCAGTACAGTTCCGGGGGGTTGTATGATGTACTGTTTTGTTTCTACTCAACCAGAGAGGG ATGACTACTTTTCTGCCGCTCCTCCAGAACAACGAATTATTATAATGTACTATAATGATACAATCGTGGAGCGCAT AATTAATCCACCCGGGGTACTAGATGTATGGGCAACATTGAACCCAGGAACAGGAAGCGGGGTATATTATTTAGGT TGGGTGCTCTTTCCAATATATGGCGGCGTGATTAAAGGTACGAGTTTGGAATAATCAAGCAAATAAATACTTA TCCCCCAGATGGTTGCTGCTCTCTGCTCACAAAACCAGGCAACTCAAGTCCAAAATGCTAAGTCATCATACTATAG CAGCTGGTTTGGCAATCGAATGATTCAGTCTGGGATCCTGGCATGTCCTCTTCGACAGGATCTAACCAATGAGTGT TTAGTTCTGCCCTTTTCTAATGATCAGGTGCTTATGGGTGCTGAAGGGAGATTATACATGTATGGTGACTCGGTGT ATTACTATCAAAGAAGCAATAGTTGGTGGCCTATGACCATGCTGTATAAGGTAACCATAACATTCACTAATGGTCA GCCATCTGCTATATCAGCTCAGAATGTGCCCACACAGCAGGTCCCTAGACCTGGGACAGGAGACTGCTCTGCAACC AATAGATGTCCCGGTTTTTGCTTGACAGGAGTGTATGCCGATGCCTGGTTACTGACCAACCCTTCGTCTACCAGTA CATTTGGATCAGAAGCAACCTTCACTGGTTCTTATCTCAACACAGCAACTCAGCGTATCAATCCGACGATGTATAT CGCGAACAACACACAGATCATAAGCTCACAGCAATTTGGATCAAGCGGTCAAGAAGCAGCATATGGCCACACAACT TGTTTTAGGGACACAGGCTCTGTTATGGTATACTGTATCTATATTATTGAATTGTCCTCATCTCTCTTAGGACAAT TTCAGATTGTCCCATTTATCCGTCAGGTGACACTATCCTAAAGGCAGAAGCCTTCAGGTCTGACCCAGCCAATCAA AGCATTATACCAGACCATGGAATGCATACCAAACATTATTGACACTAATGACACACAAAATTGGTTTTAAGAAAAA CCAAGAGAACAATAGGCCAGAATGGCTGGGTCTCGGGAGATATTACTCCCTGAAGTCCATCTCAATTCACCAATTG TAAAGCATAAGCTATACTATTACATTCTACTTGGAAACCTCCCAAATGAGATCGACCTTGACGATTTAGGTCCATT ACATAATCAAAATTGGAATCAGATAGCACATGAAGAGTCTAACTTAGCTCAACGCTTGGTAAATGTAAGAAATTTT CTAATTACCCACATCCCTGATCTTAGAAAGGGCCATTGGCAAGAGTATGTCAATGTAATACTGTGGCCGCGAATTC TTCCCTTGATCCCGGATTTTAAAATCAATGACCAATTGCCTCTGCTCAAAAATTGGGACAAGTTAGTTAAAGAATC ATGTTCAGTAATCAATGCAGGTACTTCCCAGTGCATTCAGAATCTCAGCTATGGACTGACAGGTCGTGGGAACCTC TTTACACGATCACGTGAACTCTCTGGTGACCGCAGGGATATTGATCTTAAGACAGTTGTGGCAGCATGGCATGACT CAGACTGGAAAAGAATAAGTGATTTTTGGATTATGATCAAATTCCAGATGAGACAATTAATTGTTAGGCAAACAGA TCATAATGATTCTGATTTAATCACGTATATCGAAAATAGAGAAGGCATAATCATCATAACCCCTGAACTGGTAGCA TTATTTAACACTGAGAATCATACACTAACATACATGACCTTTGAAATTGTACTGATGGTTTCAGATATGTACGAAG GTCGTCACAACATTTTATCACTATGCACAGTTAGCACTTACCTGAATCCTCTGAAGAAAAGAATAACATATTTATT GAGCCTTGTAGATAACTTAGCTTTTCAGATAGGTGATGCTGTATATAACATAATTGCTTTGCTAGAATCCTTTGTA TATGCACAGTTGCAAATGTCAGATCCCATCCCAGAACTCAGAGGACAATTCCATGCATTCGTATGTTCTGAGATTC TTGATGCACTAAGAGGAACTAATAGTTTCACCCAGGATGAATTAAGAACTGTGACAACTAATTTGATATCCCCATT CCAAGATCTGACCCCAGATCTTACGGCTGAATTGCTCTGTATAATGAGGCTTTGGGGACACCCCATGCTCACTGCC AGTCAAGCTGCAGGAAAGGTACGCGAGTCTATGTGTGCTGGAAAAGTATTAGACTTTCCCACCATTATGAAAACAC TAGCCTTTTTCCATACTATTCTGATCAATGGATACAGGAGGAAGCATCATGGAGTATGGCCACCCTTAAACTTACC GGGTAATGCTTCAAAGGGTCTCACGGAACTTATGAATGACAATACTGAAATAAGCTATGAATTCACACTTAAGCAT TGGAAGGAAGTCTCTCTTATAAAATTCAAGAAATGTTTTGATGCAGACGCAGGTGAGGAACTCAGTATATTTATGA AAGATAAGGCAATTAGTGCCCCAAAACAAGACTGGATGAGTGTGTTTTAGAAGAAGCCTAATCAAACAGCGCCATCA GCATCATCAGGTCCCCCTCACCAAATCCATTCAATCGACGGCTGTTGCTAAACTTTCTCGGAGATGACAAATTCGAC CCGAATGTGGAGCTACAGTATGTAACATCAGGTGAGTATCTACATGATGACACGTTTTGTGCATCATATTCACTAA AAGAGAAGGAAATTAAACCTGATGTGTCGAATTTTTGCAAAGTTGACTAAGAGAATGAGATCATGTCAAGTTATAGC AGAATCTCTTTTAGCGAACCATGCTGGGAAGTTAATGAAAGAGAATGGTGTTGTGATCAATCAGCTATCATTAACA AAATCACTATTAACAATGAGTCAGATTGGAATAATATCCGAGAAAGCTAGAAAGTCAACTCGAGATAACATAAATC AACCTGGTTTCCAGAATATCCAGAGAAATAAATCACATCACTCCAAGCAAGTCAATCAGCGAGATCCAAGTGATGA CTTTGAATTGGCAGCATCTTTTTTAACTACTGATCTCAAAAATATTGTTTACAATGGAGGTACCAGACAATTATC CCATTTGCTCAATCATTAAACAGAATGTATGGTTATCCTCATCTCTTTGAGTGGATTCACTTACGGCTAATGCGTA GTACACTTTACGTGGGGGATCCCTTCAACCCACCAGCAGATACCAGTCAATTTGATCTAGATAAAGTAATTAATGG AGATATCTTCATTGTATCACCCAGAGGTGGAATTGAAGGGCTGTGTCAAAAGGCTTGGACAATGATATCTATCGCT GTGATAATTCTATCTGCCACAGAGTCTGGCACACGAGTAATGAGTATGGTGCAGGGAGATAATCAAGCAATTGCTG TCACCACACGAGTACCAAGGAGCCTGCCGACTCTTGAGAAAAAGACTATTGCTTTTAGATCTTGTAATCTATTCTT TGAGAGGTTAAAATGTAATAATTTTGGATTAGGTCACCATTTGAAAGAACAAGAGACTATCATTAGTTCTCACTTC TTTGTTTATAGCAAGAGAATATTCTATCAGGGGAGGATTCTAACGCAAGCCTTAAAAAATGCTAGTAAGCTCTGCT TGACAGCTGATGTCCTAGGAGAATGCACCCAATCATCATGTTCTAATCTTGCAACTACTGTCATGAGGTTAACTGA GAATGGTGTTGAAAAAGATATCTGTTTCTACTTGAATATCTATATGACCATCAAACAGCTCTCCTATGATATCATC TTCCCTCAAGTGTCAATTCCTGGAGATCAGATCACATTAGAATACATAAATAATCCACACCTGGTATCACGATTGG CTCTTTTGCCATCCCAGTTAGGAGGTCTAAACTACCTGTCATGCAGTAGGCTGTTCAATCGAAACATAGGCGACCC GGTGGTTTCCGCAGTTGCAGATCTTAAGAGATTAATTAAATCAGGATGTATGGATTACTGGATCCTTTATAACTTA TTAGGGAGAAAACCGGGAAACGGCTCATGGGCTACTTTAGCAGCTGACCCGTACTCAATCAATATAGAGTATCAAT ACCCTCCAACTACAGCTCTTAAGAGGCACACCCAACAAGCTCTGATGGAACTCAGTACGAATCCAATGTTACGTGG CATATTCTCTGACAATGCACAGGCAGAAGAAAATAACCTTGCTAGGTTTCTCCTGGATAGGGAGGTGATCTTTCCG CGTGTAGCTCACATCATCATTGAGCAAACCAGTGTCGGGAGGAGAAAACAGATTCAAGGATATTTGGATTCAACTA GATCGATAATGAGGAAATCACTAGAAATTAAGCCCTTATCCAATAGGAAGCTTAATGAAATACTGGATTACAACAT CAATTACCTAGCTTACAATTTGGCATTACTCAAGAATGCTATTGAACCTCCGACTTATTTGAAGGCAATGACACTT GAAACATGTAGCATCGACATTGCAAGGAACCTCCGGAAGCTCTCCTGGGCCCCACTCTTGGGTGGGAGAAATCTTG AAGGATTAGAGACGCCAGATCCCATTGAAATTACTGCAGGAGCATTAATTGTTGGATCGGGCTACTGTGAACAGTG TGCTGCAGGAGACAATCGATTCACATGGTTTTTCTTGCCATCTGGTATCGAGATAGGAGGGGATCCCCGTGATAAT CCTCCTATCCGTGTACCGTACATTGGCTCCAGGACTGATGAGAGGAGGGTAGCCTCAATGGCATACATCAGGGGTG CCTCGAGTAGCTTAAAAGCAGTTCTTAGACTGGCGGGAGTGTACATCTGGGCATTCGGAGATACTCTGGAGAATTG GATAGATGCACTGGATTTGTCTCACACTAGAGTTAACATCACACTTGAACAGCTGCAATCCCTCACCCCACTTCCA ACCTCTGCCAATCTAACCCATCGGTTGGATGATGGCACAACTACCCTAAAGTTTACTCCTGCGAGCTCTTACACCT TTTCAAGTTTCACTCATATATCAAATGATGAGCAATACCTGACAATTAATGACAAAACTGCAGATTCAAATATAAT CTACCAACAGTTAATGATCACTGGACTCGGAATCTTAGAAACATGGAATAATCCCCCAATCAATAGAACATTCGAA GAATCTACCCTACATTTGCACACTGGTGCATCATGTTGTGTCCGACCTGTGGACTCCTGCATTCTCTCAGAAGCAT TAACAGTCAAGCCACATATTACAGTACCGTACAGCAATAAATTTGTATTTGATGAGGACCCGCTATCTGAATATGA AACTGCAAAACTGGAATCGTTATCATTCCAAGCCCAATTAGGCAACATTGATGCTGTAGATATGACAGGTAAATTA ACATTATTGTCCCAATTCACTGCAAGGCAGATTATCAATGCAATCACTGGACTCGATGAGTCTGTCTCTCTTACTA ATGATGCCATTGTTGCATCAGACTATGTCTCCAATTGGATTAGTGAATGCATGTATACCAAATTAGATGAATTATT TATGTATTGTGGGTGGGAACTACTATTGGAACTATCCTATCAAATGTATTATCTGAGGGTAGTTGGGTGGAGTAAT ATAGTGGATTATTCTTACATGATCTTGAGAAGAATCCCGGGTGCAGCATTAAACAATCTGGCATCTACATTAAGTC ATCCAAAACTTTTCCGACGAGCTATCAACCTAGATATAGTTGCCCCCTTAAATGCTCCTCATTTTGCATCTCTGGA CTACATCAAGATGAGTGTGGATGCAATACTCTGGGGCTGTAAAAGAGTCATCAATGTGCTCTCCAATGGAGGGGAC TTAGAATTAGTTGTGACATCTGAAGATAGCCTTATTCTCAGTGACCGATCCATGAATCTCATTGCAAGGAAATTAA CTTTATTATCACTGATTCACCATAATGGTTTGGAACTACCAAAGATTAAGGGGTTCTCTCCTGATGAGAAGTGTTT CGCTTTGACAGAATTTTTGAGGAAAGTGGTGAACTCAGGGTTGAGTTCAATAGAGAACCTATCAAATTTTATGTAC AATGTGGAGAACCCACGGCTTGCAGCATTCGCCAGCAACAATTACTACCTGACCAGAAAATTATTGAATTCAATAC GAGATACTGAGTCGGGTCAAGTAGCAGTCACCTCATATTATGAATCATTAGAATATATTGATAGTCTTAAGCTAAC CCCACATGTGCCTGGCACCTCATGCATTGAGGATGATAGTCTATGTACAAATGATTACATAATCTGGATCATAGAG TCTAATGCAAACTTGGAGAAGTATCCAATTCCAAATAGCCCTGAGGATGATTCCAATTTCCATAACTTTAAGTTGA ATGCTCCATCGCACCATACCTTACGCCCATTAGGGTTGTCATCAACTGCTTGGTATAAGGGTATAAGCTGCTGCAG GTACCTTGAGCGATTAAAGCTACCACAAGGTGATCATTTATATATTGCAGAAGGTAGTGGTGCCAGTATGACAATC ATAGAATACCTATTCCCAGGAAGAAAGATATATTACAATTCTTTATTTAGTAGTGGTGACAATCCCCCACAAAGAA ATTATGCACCAATGCCTACTCAGTTCATTGAGAGTGTCCCATACAAGCTCTGGCAAGCACACACAGATCAATATCC CGAGATTTTTGAGGACTTCATCCCTCTATGGAACGGAAACGCCGCCATGACTGACATAGGAATGACAGCTTGTGTA GAATTCATCATCAATCGAGTCGGCCCAAGGACTTGCAGTTTAGTACATGTAGATTTGGAATCAAGTGCAAGCTTAA ATCAACAATGCCTGTCAAAGCCGATAATTAATGCTATCATCACTGCTACAACTGTTTTGTGCCCTCATGGGGTGCT TATTCTGAAATATAGTTGGTTGCCATTTACTAGATTTAGTACTTTGATCACTTTCTTATGGTGCTACTTTGAGAGA ATCACTGTTCTTAGGAGCACATATTCTGATCCAGCTAATCATGAGGTTTATTTAATTTGTATCCTTGCCAACAACT TTGCATTCCAGACTGTCTCGCAGGCAACAGGAATGGCGATGACTTTAACTGATCAAGGGTTTACTTTGATATCACC TGAAAGAATAAATCAGTATTGGGATGGTCACTTGAAGCAAGAACGTATCGTAGCAGAAGCAATTGATAAGGTGGTT CTAGGAGAAAATGCTCTATTTAATTCGAGTGATAATGAATTAATTCTCAAATGTGGAGGGACACCAAATGCACGGA ATCTCATCGATATCGAGCCAGTCGCAACTTTCATAGAATTTGAACAATTGATCTGCACAATGTTGACAACCCACTT GAAGGAAATAATTGATATAACAAGGTCTGGAACCCAGGATTATGAAAGTTTATTACTCACTCCTTACAATTTAGGT CTTCTTGGTAAAATCAGTACGATAGTGAGATTATTAACAGAAAGGATTCTAAATCATACTATCAGGAATTGGTTGA TCCTCCCACCTTCGCTCCGGATGATCGTGAAGCAGGACTTGGAATTCGGCATATTCAGGATTACTTCCATCCTCAA TTCTGATCGGTTCCTGAAGCTTTCTCCAAATAGGAAATACTTGATTGCACAATTAACTGCAGGCTACATTAGGAAA TTGATTGAGGGGGATTGCAATATCGATCTAACCAGACCTATCCAAAAGCAAATCTGGAAAGCATTAGGTTGTGTAG TCTATTGTCACGATCCAATGGATCAAAGGGAGTCAACAGAGTTTATTGATATAAATATTAATGAAGAAATAGACCG CGGGATCGATGGCGAGGAAATCTAAACATATCAAGAATCAGAATTAGTTTAAGAAAAAAGAAGAGGATTAATCTTG GTTTTCCCCTTGGT

[0171] The underlined sequence is the PIV5-CVB genomic nucleic acid sequence. The italicized and underlined sequences are the entire MOMP nucleic acid sequence. The double-underlined sequence is the AOS-modified NT-MOMPΔsp nucleic acid sequence.

[0172] c. pMHD116-CVB-NTTM-OspA BPBPk

[0173] The pMHD116-CVB-NTTM-OspA provided in this article BPBPk The nucleic acid sequences from 5' to 3' are as follows:

[0174] ACCAAGGGGAAAATGAAGTGGTGACTCAAATCATCGAAGACCCTCGAGATTACATAGGTCCGGAACCTA TGGCCTTCGTGACCGACCTCGAGTCAGAGTAGTTCAATAAGGACCTATCAAGTTTGGGCAATTTTTCGTCCCCGACA CAAAAATGTCATCCGTGCTTAAAGCATATGAGCGATTCACGCTCACTCAAGAACTGCAAGATCAGAGTGAGGAAGGT ACAATCCCACCTACAACACTAAAACCGGTAATCAGGGTATTTATACTAACCTCTAATAACCCAGAGCTAAGATCCCG GCTTCTTCTATTCTGCCTACGGATTGTTCTCAGTAATGGTGCAAGGGATTCCCATCGCTTTGGAGCATTACTCACAA TGTTTTCGCTACCATCAGCCACAATGCTCAATCATGTCAAATTAGCTGACCAGTCACCAGAAGCTGATATCGAAAGG GTAGAGATCGATGGCTTTGAGGAGGGATCATTCCGCTTAATCCCCAATGCTCGTTCAGGTATGAGCCGTGGAGAGAT CAATGCCTATGCTGCACTTGCAGAAGATCTACCTGACACACTAAACCATGCAACACCTTTCGTTGATTCCGAAGTCG AGGGAACTGCATGGGATGAGATTGAGACTTTCTTAGATATGTGTTACAGTGTCCTAATGCAGGCATGGATAGTGACT TGCAAGTGCATGACTGCGCCAGACCAACCTGCTGCTTCTATTGAGAAACGCCTGCAAAAATATCGTCAGCAAGGCAG GATCAACCCGAGATATCTCCTGCAACCGGAGGCTCGACGAATAATCCAGAATGTAATCCGGAAGGGAATGGTGGTCA GACATTTCCTCACCTTTGAACTGCAGCTTGCCCGAGCACAAAGCCTTGTATCAAATAGGTATTATGCTATGGTAGGG GATGTTGGAAAGTATATAGAGAATTGTGGAATGGGAGGCTTCTTTTTGACACTAAAATATGCATTAGGAACTAGATG GCCCACACTTGCTTTAGCTGCATTTTCAGGAGAGCTAACAAAGCTAAAGTCCCTCATGGCATTATACCAGACCCTTG GTGAGCAGGCCCGATATTTGGCCCTATTGGAGTCACCACATTTGATGGATTTTGCTGCAGCAAACTACCCACTGCTA TATAGCTATGCTATGGGAATAGGCTATGTGTTAGATGTCAACATGAGGAACTACGCTTTCTCCAGATCATACATGAA CAAGACATATTTCCAATTGGGAATGGAAACTGCAAGAAAACAACAGGGTGCAGTTGACATGAGGATGGCAGAAGATC TCGGTCTAACTCAAGCCGAACGCACCGAGATGGCAAATACACTTGCCAAATTGACCACAGCAAATCGAGGGGCAGAC ACCAGGGGAGGAGTCAACCCGTTCTCATCTGTCACTGGGACAACTCAGGTGCCCGCTGCAGCAACAGGTGACACACT CGAGAGTTACATGGCAGCGGATCGACTGAGGCAGAGATATGCTGATGCAGGCACCCATGATGATGAGATGCCACCAT TGGAAGAGGAGGAAGAGGACGACACATCTGCAGGTCCACGCACTGGACCAACTCTTGAACAAGTGGCCTTGGACATC CAGAACGCAGCAGTTGGAGCTCCCATCCATACAGATGACCTGAATGCCGCACTGGGTGATCTTGACATCTAGACAAT TCAGATCCCAATCTAAAATTGACATACCTAATTGATTAGTTAGATGGAACTACAGTGGATTCCATAAGGTTCCTGCC TACCATCGGCTTTAAAGAAAAAAATAGGCCCGGACGGGTTAGCAACAAGCGACTGCCGGTGCCAACAGCGCAATCCA CAATCTACAATGGATCCCACTGATCTGAGCTTCTCCCCAGATGAGATCAATAAGCTCATAGAGACAGGCCTGAATAC TGTAGAGTATTTTACTTCCCAACAAGTCACAGGAACATCCTCTCTTGGAAAGAATACAATACCACCAGGGGTCACAG GACTACTAACCAATGCTGCAGAGGCAAAGATCCAAGAGTCAACTAACCATCAGAAGGGCTCAGTTGGTGGGGGTGCA AAACCAAAGAAACCGCGACCAAAAATTGCCATTGTGCCAGCAGATGACAAAACAGTGCCCGGAAAGCCGATCCCAAA CCCTCTATTAGGTCTGGACTCCACCCCGAGCACCCAAACTGTGCTTGATCTAAGTGGGAAAACATTACCATCAGGAT CCTATAAGGGGGTTAAGCTTGCGAAATTTGGAAAAGAAAATCTGATGACACGGTTCATCGAGGAACCCAGAGAGAAT CCTATCGCAACCAGTTTCCCCATCGATTTTAAGAGGGGCAGGGATACCGGCGGGTTCCATAGAAGGGAGTACTCAAT CGGATGGGTGGGAGATGAAGTCAAGGTCACTGAGTGGTGCAATCCATCCTGTTCTCCAATCACCGCTGCAGCAAGGC GATTTGAATGCACTTGTCACCAGTGTCCAGTCACTTGCTGAATGTGAACGAGATACTTAATACAGTGAGAAATTT GGACTCTCGGATGAATCAACTGGAGACAAAAGTAGATCGCATTCTCTCATCTCAGTCTAATCCAGACCATCAAGA ATGACATAGTTGGACTTAAAGCAGGGATGGCTACTTTAGAAGGAATGATTACAACTGTGAAAATCATGGACCCGGGA GTTCCCAGTAATGTTACTGTGGAAGATGTACGCAAGACACTAAGTAACCATGCTGTTGTTGTGCCAGAATCATTCAA TGATAGTTTCTTGACTCAATCTGAAGATGTAATTTCACTTGATGAGTTGGCTCGACCAACTGCAACAAGTGTTAAGA AGATTGTCAGGAAGGTTCCTCCTCAGAAGGATCTGACTGGATTGAAGATTACACTAGAGCAATTGGCAAAGGATTGC ATCAGCAAACCGAAGATGAGGGAAGAGTATCTCCTCAAAATCAACCAGGCTTCCAGTGAGGCTCAGCTAATTGACCT CAAGAAAGCAATCATCCGCAGTGCAATTTGATCAAGAAACACCCAATTACACTACACTGGTATGACACTGTACTAAC CCTGAGGGTTTTAGAAAAAACGATTAACGATAAATAAGCCCGAACACTACACACTACCTGAGGCAGCCATGCCATCC ATCAGCATTCCCGCAGACCCCACCAATCCACGTCAATCAATAAAAGCGTTCCCAATTGTGATCAACAGTGATGGGGG TGAGAAAGGCCGCTTGGTTAAACAACTACGCACAACCTACTTGAATGACCTAGATACTCATGAGCCACTGGTGACAT TCATAAATACCTATGGATTCATCTACGAACAGGATCGGGGGAATACCATTGTCGGAGAGGATCAACTTGGGAAGAAA AGAGAGGCTGTGACCGCTGCAATGGTTACCCTTGGATGTGGGCCTAATCTACCATCATTAGGGAATGTCCTGGGACA ACTGAGGGAATTCCAGGTCACTGTTAGGAAGACATCCAGCAAAGCGGAAGAGATGGTCTTTGAAATTGTTAAGTATC CGAGAATATTTCGGGGTCATACATTAATCCAGAAAGGACTAGTCTGTGTCTCCGCAGAAAAATTTGTTAAGTCACCA GGGAAAATACAATCTGGAATGGACTATCTCTTCATTCCGACATTTCTGTCAGTGACTTACTGTCCAGCTGCAATCAA ATTTCAGGTACCTGGCCCCATGTTGAAAATGAGATCAAGATACACTCAGAGCTTACAACTTGAACTAATGATAAGAA TCCTGTGTAAGCCCGATTCGCCACTTATGAAGGTCCATACCCCTGACAAGGAGGGAAGAGGATGTCTTGTATCAGTA TGGCTGCATGTATGCAACATCTTCAAATCAGGAAACAAGAATGGCAGTGAGTGGCAGGAATACTGGATGAGAAAGTG TGCTAACATGCAACTTGAAGTGTCGATTGCAGATATGTGGGGACCAACTATCATAATTCATGCCAGAGGTCACATTC CCAAAAGTGCTAAGTTGTTTTTTGGAAAGGGTGGATGGAGCTGCCATCCACTTCACGAAGTTGTTCCAAGTGTCACT AAAACACTATGGTCCGTGGGCTGTGAGATTACAAAGGCGAAGGCAATAATACAAGAGAGTAGCATCTCTCTTCTCGT GGAGACTACTGACATCATAAGTCCAAAAGTCAAAATTTCATCTAAGCATCGCCGCTTTGGGAAATCAAATTGGGGTC TGTTCAAGAAAACTAAATCACTGCCTAACCTGACGGAGCTGGAATGACTGACCTCTAATCGAGACTACACCGCCGCA AACTATAGGTGGGTGGTACCTCAGTGATTAATCTTGTAAGCACTGATCGTAGGCTACAACACACTAATATTATCCAG ATTAGAGAGCTTAATTAGCTCTGTATTAATAATAACACTACTATTCCAATAACTGGAATCACCAGCTTGATTTATCT CCAAAATGATTCAAAGAAAACAAATCATATTAAGACTATCCTAAGCACGAACCCATATCGTCCTTCAAATCATGGGT ACTATAATTCAATTTCTGGTGGTCTCCTGTCTATTGGCAGGAGCAGGCAGCCTTGATCCAGCAGCCCTCATGCAAAT CGGTGTCATTCCAACAAATGTCCGGCAACTTATGTATTATACTGAAGCTTCATCAGCATTCATTGTTGTGAAGTTAA TGCCTACAATTGACTCGCCGATTAGTGGATGTAATATAACATCAATTTCAAGCTATAATGCAACAGTGACAAAACTC CTACAGCCGATCGGTGAGAATTTGGAGACGATTAGGAACCAGTTGATTCCAACTCGGAGGAGACGCCGGTTTGCAGG GGTGGTGATTGGATTAGCTGCATTAGGAGTAGCTACTGCCGCACAGGTCACTGCCGCAGTGGCACTAGTAAAGGCAA ATGAAAATGCTGCGGCTATACTCAATCTCAAAAATGCAATCCAAAAAACAAATGCAGCAGTTGCAGATGTGGTCCAG GCCACACAATCACTAGGAACGGCAGTTCAAGCAGTTCAAGATCACATAAACAGTGTGGTAAGTCCAGCAATTACAGC AGCCAATTGTAAGGCCCAAGATGCTATCATTGGCTCAATCCTCAATCTCTATTTGACCGAGTTGACAACCATCTTCC ACAATCAAATTACAAACCCTGCATTGAGTCCCATTACAATTCAAGCTTTAAGGATCCTACTGGGGAGTACCTTGCCG ACTGTGGTCGAAAAATCTTTCAATACCCAGATAAGTGCAGCTGAGCTTCTCTCATCAGGGTTATTGACAGGCCAGAT TGTGGGATTAGATTTGACCTATATGCAGATGGTCATAAAAATTGAGCTGCCAACTTTAACTGTACAACCTGCAACCC AGATCATAGATCTGGCCACCATTTCTGCATTCATTAACAATCAAGAAGTCATGGCCCAATTACCAACACGTGTTATG GTGACTGGCAGCTTGATCCAAGCCTATCCCGCATCGCAATGCACCATTACACCCAACACTGTGTACTGTAGGTATAA TGATGCCCAAGTACTCTCAGATGATACTATGGCTTGCCTCCAAGGTAACTTGACAAGATGCACCTTCTCTCCAGTGG TTGGGAGCTTTCTCACTCGATTCGTGCTGTTCGATGGAATAGTTTATGCAAATTGCAGGTCGATGTTGTGCAAGTGC ATGCAACCTGCTGCTGTGATCCTACAGCCGAGTTCATCCCCTGTAACTGTCATTGACATGTACAAATGTGTGAGTCT GCAGCTTGACAATCTCAGATTCACCATCACTCAATTGGCCAATGTAACCTACAATAGCACCATCAAGCTTGAATCAT CCCAGATCTTGTCTATTGATCCGTTGGATATATCCCAGAATCTAGCTGCGGTGAATAAGAGTCTAAGTGATGCACTA CAACACTTAGCACAAAGTGACACATATCTTTCTGCAATCACATCAGCTACGACTACAAGTGTATTATCCATAATAGC AATCTGTCTTGGATCGTTAGGTTTAATATTAATAATCTTGCTCAGTGTAGTTGTGTGGAAGTTATTGACCATTGTCG TTGCTAATCGAAATAGAATGGAGAATTTTGTTTATCATAAATAAGCATTCCACCACTCACGATCTGATCTCAGTGAG AAAAATCAACCTGCAACTCTTGGAACAAGATAAGACAGTCATCCATTAGTAATTTTTAAGAAAAAAACGATAGGACC GAACCTAGTATTGAAAGAACCGTCTCGGTCAATCTAGGTAATCGAGCTGATACCGTCTCGGAAAGCTCAAGCGGCC

[0175]

[0176] GGCATCGGCCTGATCCTGGCCCTGATCGCCTGCAAGCAGAACGTGAGCTCCCTGGACGAGAAGAATAG CGTGTCCGTGGATCTGCCCGGCGAGATGAAGGTGCTGGTGAGCAAGGAGAAGAACAAGGACGGCAAGTATGATCTG ATCGCCACAGTGGACAAGCTGGAGCTGAAGGGCACCAGCGATAAGAACAATGGATCCGGCGTGCTGGAGGGAGTGA AGGCAGACAAGTCTAAGGTGAAGCTGACCATCAGCGACGATCTGGGCCAGACCACACTGGAGGTGTTCAAGGAGGA CGGCAAGACCCTGGTGTCCAAGAAGGTGACATCTAAGGATAAGTCTAGCACCGAGGAGAAGTTTAACGAGAAGGGC GAGGTGTCCGAGAAGATCATCACACGGGCCGACGGCACCAGACTGGAGTACACAGGCATCAAGTCCGATGGCTCTG GCAAGGCCAAGGAGGTGCTGAAGGGCTTCACACTGGAGGGCAAGGTGGCCAATGATAAGGTGACCCTGGAGGTGAA GGAGGGCACCGTGACACTGTCCAAGAACATCTCTAAGAGCGGCGAGGTGTCTGTGGAGCTGAATGACACCGATTCC TCTGCCGCCACAAAGAAGACCGCCGCCTGGAACAGCAAGACCTCCACACTGACCATCTCTGTGAATAGCAAGAAGA CCACACAGCTGGTGTTTACAAAGCAGGACACAATCACCGTGCAGAAGTATGATAGCGCCGGAACCAACCTGGAGGG AACAGCAGTGGAGATCAAGACCCTGGACGAGCTGAAGAATGCCCTGAAGTGAGTCGACCACCTGCTATAGGCTATC CACTGCATCATCTCTCCTGCCATACTTCCTACTCACATCATATCTATTTTAAAGAAAAAATAGGCCCGAACACTAA TCGTGCCGGCAGTGCCACTGCACACACAACACTACACATACAATACACTACAATGGTTGCAGAAGATGCCCCTGTT AGGGCCACTTGCCGAGTATTATTTCGAACAACAACTTTAATCTTTCTATGCACACTACTAGCATTAAGCATCTCTA TCCTTTATGAGAGTTTAATAACCCAAAAGCAAATCATGAGCCAAGCAGGCTCAACTGGATCTAATTCTGGATTAGG AAGTATCACTGATCTTCTTAATAATATTCTCTCTGTCGCAAATCAGATTATATATAACTCTGCAGTCGCTCTACCT CTACAATTGGACACTCTTGAATCAACACTCCTTACAGCCATTAAGTCTCTTCAAACCAGTGACAAGCTAGAACAGA ACTGCTCGTGGAGTGCTGCACTGATTAATGATAATAGATACATTAATGGCATCAATCAGTTCTATTTTTCAATTGC TGAGGGTCGCAATCTGACACTTGGCCCACTTCTTAATATGCCTAGTTTCATTCCAACTGCCACGACACCAGAGGGC TGCACCAGGATCCCATCATTCTCGCTCACTAAGACACACTGGTGTTATACACACAATGTTATCCTGAATGGATGCC AGGATCATGTATCCTCAAATCAATTTGTTTCTATGGGAATCATTGAACCCACTTCTGCCGGGTTTCCATTCTTTCG AACCCTAAAGACTCTATATCTCAGCGATGGGGTCAATCGTAAGAGCTGCTCTATCAGTACAGTTCCGGGGGGTTGT ATGATGTACTGTTTTGTTTCTACTCAACCAGAGAGGGATGACTACTTTTCTGCCGCTCCTCCAGAACAACGAATTA TTATAATGTACTATAATGATACAATCGTGGAGCGCATAATTAATCCACCCGGGGTACTAGATGTATGGGCAACATT GAACCCAGGAACAGGAAGCGGGGTATATTATTTAGGTTGGGTGCTCTTTCCAATATATGGCGGCGTGATTAAAGGT ACGAGTTTATGGAATAATCAAGCAAATAAATACTTTATCCCCCAGATGGTTGCTGCTCTCTGCTCACAAAACCAGG CAACTCAAGTCCAAAATGCTAAGTCATCATACTATAGCAGCTGGTTTGGCAATCGAATGATTCAGTCTGGGATCCT GGCATGTCCTCTTCGACAGGATCTAACCAATGAGTGTTTAGTTCTGCCCTTTTCTAATGATCAGGTGCTTATGGGT GCTGAAGGGAGATTATACATGTATGGTGACTCGGTGTATTACTATCAAAGAAGCAATAGTTGGTGGCCTATGACCA TGCTGTATAAGGTAACCATAACATTCACTAATGGTCAGCCATCTGCTATATCAGCTCAGAATGTGCCCACACAGCA GGTCCCTAGACCTGGGACAGGAGACTGCTCTGCAACCAATAGATGTCCCGGTTTTTGCTTGACAGGAGTGTATGCC GATGCCTGGTTACTGACCAACCCTTCGTCTACCAGTACATTTGGATCAGAAGCAACCTTCACTGGTTCTTATCTCA ACACAGCAACTCAGCGTATCAATCCGACGATGTATATCGCGAACAACACACAGATCATAAGCTCACAGCAATTTGG ATCAAGCGGTCAAGAAGCAGCATATGGCCACACAACTTGTTTTTAGGGACACAGGCTCTGTTATGGTATACTGTATC TATATTATTGAATTGTCCTCATCTCTCTTAGGACAATTTCAGATTGTCCCATTTATCCGTCAGGTGACACTATCCT AAAGGCAGAAGCCTTCAGGTCTGACCCAGCCAATCAAAGCATTATACCAGACCATGGAATGCATACCAAACATTAT TGACACTAATGACACACAAAATTGGTTTTAAGAAAACCAAGAGAACAATAGGCCAGAATGGCTGGGTCTCGGGAG ATATTACTCCCTGAAGTCCATCTCAATTCACCAATTGTAAAGCATAAGCTATACTATTACATTCTACTTGGAAACC TCCCAAATGAGATCGACCTTGACGATTTAGGTCCATTACATAATCAAAATTGGAATCAGATAGCCACATGAAGAGTC TAACTTAGCTCAACGCTTGGTAAATGTAAAAATTTCTAATTACCCACATCCCTGATCTTAGAAAGGGCCATTGG CAAGAGTATGTCAATGTAATACTGTGGCCGCGAATTCTTCCCTTGATCCCGGATTTTAAAATCAATGACCAATTGC CTCTGCTCAAAAATTGGGACAAGTTAGTTAAAAGAATCATGTTCAGTAATCAATGCAGGTACTTCCCAGTGCATTCA GAATCTCAGCTATGGACTGACAGGTCGTGGGAACCTCTTTACACGATCACGTGAACTCTCTGGTGACCGCAGGGAT ATTGATCTTAAGACAGTTGTGGCAGCATGGCATGACTCAGACTGGAAAAGAATAAGTGATTTTTGGATTATGATCA AATTCCAGATGAGACAATTAATTGTTAGGCAAACAGATCATAATGATTCTGATTTAATCACGTTATATCGAAAATAG AGAAGGCATAATCATCATAACCCCTGAACTGGTAGCATTATTTAACACTGAGAATCATACACTAACATACATGACCTTTGAAATTGTACTGATGGTTTCAGATATGTACGAAGGTCGTCACAACATTTTCACTATGCACAGTTAGCACTT ACCTGAATCCCTCTGAAGAAAAGAATAACATATTTATTGAGCCTTGTAGATAACTTAGCTTTTCAGATAGGTGATGC TGTATAACATAATTGCTTTGCTAGAATCCTTTGTATATGCACAGTTGCAAATGTCAGATCCCATCCCAGAACTC AGAGGACAATTCCATGCATTCGTATGTTCTGAGATTCTTGATGCACTAAGAGGAACTAATAGTTTCACCCAGGATG AATTAAGAACTGTGACAACTAATTTGATATCCCCATTCCAAGATCTGACCCCAGATCTTACGGCTGAATTGCTCTG TATAATGAGGCTTTGGGGACACCCCATGCTCACTGCCAGTCAAGCTGCAGGAAAGGTACGCGAGTCTATGTGTGCT GGAAAAGTATTAGACTTTCCCACCATTATGAAAACACTAGCCTTTTTCCATACTATTCTGATCAATGGATACAGGA GGAAGCATCATGGAGTATGGCCACCCTTAAACTTACCGGGTAATGCTTCAAAGGGTCTCACGGAACTTATGAATGA CAATACTGAAATAAGCTATGAATTCCACTTAAGCATTGGAAGGAAGTCTCTCTTATAAAATTCAAGAAATGTTTT GATGCAGACGCAGGTGAGGAACTCAGTATTTATGAAAGATAAGGCAATTAGTGCCCCAAAACAAGACTGGATGA GTGTGTTTAGAAGAAGCCTAATCAAACAGCGCCATCAGCATCATCAGGTCCCCCTACCAAAATCCATTCAATCGACG GCTGTTGCTAAACTTTCTCGGAGATGACAAATTCGACCCGAATGTGGAGCTACAGTATGTAACATCAGGTGAGTAT CTACATGATGACACGTTTTGTGCATCATTCACTAAAAGAAGGAAATTAAACCTGATGGTCGAATTTTTGCAA AGTTGACTAAGAGAATGAGATCATGTCAAGTTATAGCAGAATCTCTTTTAGCGAACCATGCTGGGAAGTTAATGAA AGAGAATGGTGTTGTGTGATGAATCAGCTATCATTAAACAAAATCACTATTAACAATGAGTCAGATTGGAATAATATCC GAGAAAGCTAGAAAGTCAACTCGAGATAACATAAATCAACCTGGTTTCCAGAAATATCCAGAGAAATAAATCACATC ACTCCAAGCAAGTCAATCAGCGAGATCCAAGTGATGACTTTGAATTGGCAGCATCTTTTTTAACTACTGATCTCAA AAAATATTGTTTACAATGGAGGTACCAGACAATTATCCCATTTGCTCAATCATTAAACAGAATGTATGGTTATCCT CATCTCTTTGAGTGGATTCACTTACGGCTAATGCGTAGTACACTTTACGTGGGGGATCCCTTCAACCCACCAGCAG ATACCAGTCAATTTGATCTAGATAAAGTAATTAATGGAGATATCTTCATTGTATCACCCAGAGGTGGAATTGAAGG GCTGTGTCAAAAGGCTTGGACAATGATATCTATCGCTGTGATAATTCTATCTGCCACAGAGTCTGGCACACGAGTA ATGAGTATGGTGCAGGGAGATAATCAAGCAATTGCTGTCACCACAGAGTACCAAGGAGCCTGCCGACTCTTGGA AAAAGACTATTGCTTTTAGATCTTGTAATCTATTCTTTGAGAGGTTAAAATGTAATAATTTTGGATTAGGTCACCA TTTGAAACAAGAGACTATCATTAGTTCTCACTTCTTTGTTTATAGCAAGAGAATATTCTATCAGGGGGAGGATT CTAACGCAAGCCTTAAAAAATGCTAGTAAGCTCTGCTTGACAGCTGATGTCCTAGGAGAATGCACCCAATCATCAT GTTCTAATCTTGCAACTACTGTCATGAGGTTAACTGAGAATGGTGTTGAAAAAGATATCTGTTTCTACTTGAATAT CTATATGACCATCAAACAGCTCTCCTATGATATCATCTTCCCTCCAAGTGTCAATTCCTGGAGATCAGATCACATTA GAATACATAAATAATCCACACCTGGTATCACGATTGGCTCTTTTGCCATCCCAGTTAGGAGGTCTAAACTACCTGT CATGCAGTAGGCTGTTCAATCGAAACATAGGCGACCCGGTGGTTTCCGCAGTTGCAGATCTAAGAGATTAATTAA ATCAGGATGTATGGATTACTGGATCCTTTATAACTTATTAGGGAGAAACCGGGAAACGGCTCATGGGCTACTTTA GCAGCTGACCCGGTACTCAATCAATATAGAGTATCAATACCCTCCAACTACAGCTCTTAAGAGGCACACCAACAAG CTCTGATGGAACTCAGTACGAATCCAATGTTACGTGGCATATTCTCTGACAATGCACAGGCAGAAGAAATAACCT TGCTAGGTTTCTCCTGGATAGGGAGGTGATCTTTCCGCGTGTAGCTCACATCATCATTGAGCAAACCAGTGTCGGG AGGAGAAAACAGATTCAAGGATATTTGGATTCAACTAGATCGATAATGAGGAAATCACTAGAAATTAAGCCCTTAT CCAATAGGAAGCTTAATGAAATACTGGATTACAACATCAATTACCTAGCTTACAATTTGGCATTACTCAAGAATGC TATTGAACCTCCGACTTATTTGAAGGCAATGACACTTGAAACATGTAGCATCGACATTGCAAGGAACCTCCGGAAG CTCTCCTGGGCCCACTCTTGGGTGGGGAAATCTTGAAGGATTAGAGACGCCAGATCCCATTGAAATTACTGCAG GAGCATTAATTGTTGGATCGGCTACTGTGAACAGTGTGCTGCAGGAGACAATCGATTCACATGGTTTTTCTTGCC ATCTGGTATCGAGATAGGAGGGATCCCCGTGATAATCCTCCTATCCGTGTACCGTACATTGGCTCCAGGACTGAT GAGAGGAGGGTAGCCTCAATGGCATACATCAGGGGTGCCTCGAGTAGCTTAAAAGCAGTTCTTAGACTGGCGGGAG TGTACATCTGGGCATTCGGAGATACTCTGGAGAATTGGATAGATGCACTGGATTTGTCTCACACTAGAGTTAACAT CACACTTGAACAGCTGCAATCCCTCACCCCACTTCCAACCTCTGCCAATCTAACCCATCGGTTGGATGATGGCACA ACTACCCTAAAGTTTACTCCTGCGAGCTCTTACACCTTTTCAAGTTTCACTCATATATCAAATGATGAGCAATACC TGACAATTAATGACAAAACTGCAGATTCAAATATAATCTACCAACAGTTAATGATCACTGGACTCGGAATCTTAGA AACATGGAATAATCCCCCAATCAATAGAACATTCGAAGAATCTACCCTACATTTGCACACTGGTGCATCATGTTGT GTCCGACCTGTGGACTCCTGCATTCTCTCAGAAGCATTAACAGTCAAGCCACATATTACAGTACCGTACAGCAATA AATTTGTATTTGATGAGGACCCGCTATCTGAATATGAAACTGCAAAACTGGAATCGTTATCATTCCAAGCCCAATT AGGCAACATTGATGCTGTAGATATGACAGGTAAATTAACATTATTGTCCCAATTCACTGCAAGGCAGATTATCAAT GCAATCACTGGACTCGATGAGTCTGTCTCTCTTACTAATGATGCCATTGTTGCATCAGACTATGTCTCCAATTGGA TTAGTGAATGCATGTATACCAAATTAGATGAATTATTTATGTATTGTGGGTGGGAACTACTATTGGAACTATCCTA TCAAATGTATTATCTGAGGGTAGTTGGGTGGAGTAATATAGTGGATTATTCTTACATGATCTTGAGAAGAATCCCG GGTGCAGCATTAAACAATCTGGCATCTACATTAAGTCATCCAAAACTTTTCCGACGAGCTATCAACCTAGATATAG TTGCCCCCTTAAATGCTCCTCATTTTGCATCTCTGGACTACATCAAGATGAGTGTGGATGCAATACTCTGGGGCTG TAAAAGAGTCATCAATGTGCTCTCCAATGGAGGGGACTTAGAATTAGTTGTGACATCTGAAGATAGCCTTATTCTC AGTGACCGATCCATGAATCTCATTGCAAGGAAATTAACTTTATTATCACTGATTCACCATAATGGTTTGGAACTAC CAAAGATTAAGGGGTTCTCTCCTGATGAGAAGTGTTTCGCTTTGACAGAATTTTTGAGGAAAGTGTGAACTCAGG GTTGAGTTCAATAGAGAACCTATCAAATTTTATGTACAATGTGGAGAACCCACGGCTTGCAGCATTCGCCAGCAAC AATTACTACCTGACCAGAAAATTATTGAATTCAATACGAGATACTGAGTCGGGTCAAGTAGCAGTCACCTCATATT ATGAATCATTAGAATATATTGATAGTCTTAAGCTAACCCCACATGTGCCTGGCACCTCATGCATTGAGGATGATAG TCTATGTACAAATGATTACATAATCTGGATCATAGAGTCTAATGCAAACTTGGAGAAGTATCCAATTCCAAATAGC CCTGAGGATGATTCCAATTTCCATAACTTTAAGTTGAATGCTCCATCGCACCATACCTTACGCCCATTAGGGTTGT CATCAACTGCTTGGTATAAGGGTATAAGCTGCTGCAGGTACCTTGAGCGATTAAAGCTACCACAAGGTGATCATTT ATATATTGCAGAAGGTAGTGGTGCCAGTATGACAATCATAGAATACCTATTCCCAGGAAGAAAGATATATTACAAT TCTTTATTTAGTAGTGGTGACAATCCCCCACAAAGAAATTATGCACCAATGCCTACTCAGTTCATTGAGAGTGTCC CATACAAGCTCTGGCAAGCACACACAGATCAATATCCCGAGATTTTTGAGGACTTCATCCCTCTATGGAACGGAAA CGCCGCCATGACTGACATAGGAATGACAGCTTGTGTAGAATTCATCATCAATCGAGTCGGCCCAAGGACTTGCAGT TTAGTACAATGTAGATTTGGAATCAAGTGCAAGCTTAAATCAACAATGCCTGTCAAAGCCGATAATTAATGCTATCA TCACTGCTACAACTGTTTTGTGCCCTCATGGGGGTGCTTATTCTGAAATATAGTTGGTTGCCATTTACTAGATTTAG TACTTTGATCACTTTCTTATGGTGCTACTTTGAGAGAATCACTGTTCTTAGGAGCACATATTCTGATCCAGCTAAT CATGAGGTTTTATTTAATTTGTATCCTTGCCAACAACTTTGCATTCCAGACTGTCTCGCAGGCAACAGGAATGGCGA TGACTTTAACTGATCAAGGGTTTACTTTGATATCACCTGAAAGAATAAATCAGTATTGGGATGGTCACTTGAAGCA AGAACGTATCGTAGCAGAAGCAATTGATAAGGTGGTTCTAGGAGAAAATGCTCTATTTAATTCGAGTGATAATGAA TTAATTCTCAAATGTGGAGGGACACCAAATGCACGGAATCTCATCCGATATCGAGCCAGTCGCAACTTTCATAGAAT TTGAACAATTGATCTGCACAATGTTGACAACCCACTTGAAGGAAATAATTGATATAACAAGGTTCGGAACCCAGGA TTATGAAAGTTTATTACTCACTCCTTACAATTTAGGTCTTCTTGGTAAAATCAGTACGATAGTGAGATTATTAACA GAAAGGATTCTAAATCATACTATCAGGAATTGGTTGATCCTCCCACCTTCGCTCCGGATGATCGTGAAGCAGGACT TGGAATTCGGCATATTCAGGATTACTTCCATCCTCAATTCTGATCGGTTCCTGAAGCTTTCTCCAAATAGGAAATA CTTGATTGCACATTAACTGCAGGCTACATTAGGAAATTGATTGAGGGGGATTGCAATATCGATCTAACCAGACCT ATCCAAAAGCAAATCTGGAAAGCATTAGGTTGTGTAGTCTATTGTCACGATCCAATGGATCAAAGGGAGTCAACAG AGTTTATTGATATAAATATTAATGAAGAAATAGACCGCGGGATCGATGGCGAGGAAATCTAAACATATCAAGAATC AGAATTAGTTTAAGAAAAAAGAAGAGGATTAATCTTGGTTTTCCCCTTGGT

[0177] The underlined sequence is the PIV5-CVB genomic nucleic acid sequence. The italicized and underlined sequences are the entire OspA nucleic acid sequence. The double-underlined sequence is AOS-modified NTTM-OspA. BPBPk Nucleic acid.

[0178] III. Preparation Method

[0179] The present invention also includes methods for preparing and using PIV5 (W3A, CPI, or CVB backbone) viral expression vectors, including but not limited to any of the methods described herein, to express modified bacterial proteins.

[0180] For example, the present invention includes a method for expressing modified bacterial proteins in cells by contacting or infecting cells with or by contacting or infecting cells with a PIV5 viral expression vector, viral particles or a composition as described herein.

[0181] This invention includes a method for inducing an immune response in a subject to a modified bacterial protein by administering to the subject a viral expression vector, viral particles, or composition as described herein. The immune response may include humoral and / or cellular immune responses. The immune response may enhance innate and / or adaptive immune responses.

[0182] The present invention includes a method for expressing heterologous modified bacterial proteins in subjects by administering viral expression vectors, viral particles or compositions as described herein.

[0183] The present invention includes a method for expressing modified bacterial proteins in a subject by administering to the subject a viral expression vector, viral particle, or composition as described herein.

[0184] IV. Treatment Methods

[0185] This disclosure can be used in gene therapy and / or therapeutic methods to treat diseases involving an increase or decrease in a target nucleotide sequence in host cells. In these embodiments, the expressible heteronucleotide sequence can be derived from a mammalian genome. In some embodiments, the expressible heteronucleotide sequence is derived from a human genome, which may be particularly useful where expression of wild-type RNA and / or proteins can produce a therapeutic effect in a patient. For example, the expressible heteronucleotide sequence may encode CFTR, NeuroD1, Cas9, and Guide RNA or any other such sequence. In other embodiments, the heteronucleotide sequence encodes a secreted protein.

[0186] In other embodiments, the expressible heteronucleotide sequence responds to a positive selection stimulus. In other embodiments, the expressible heteronucleotide sequence also responds to a negative selection stimulus. In other embodiments, it may be useful for the polynucleotide sequence to further include a reporter gene. For example, the reporter gene may be a luciferase or green fluorescent protein.

[0187] In some embodiments, PIV5 expresses one or more nucleotide sequences (e.g., siRNA) that can modify the translation and / or transcription of a host cell target nucleotide sequence within the host cell. In some embodiments, the transcription and / or translation of the expressible heteronucleotide sequence is modified to make its nucleotide sequence codonally degenerate relative to the endogenous gene in the cell. Furthermore, the expressible heteronucleotide sequence can be modified to enable the co-expression of a repressor or silencer sequence that suppresses or silences a host cell target nucleotide sequence within the host cell.

[0188] In other embodiments, the expressible target heteronucleotide produces a product that stabilizes the host cell's RNA nucleotide sequence. This product can be inducible or persistently expressed. For example, the 3′ RhoB untranslated region (UTR) can stabilize target RNAs that express toxic proteins or other target proteins in response to serum. Another example is linking the 3′ untranslated region of an eosinophil chemokine to a target gene, which typically has a short half-life, but is stabilized by adding TNF-α and IL-4 to the cell. Alternatively, sequences contained in the 16-mer sequences within the 5′ coding regions of CYP2E 1 and CYP2B 1 mRNAs destabilize the target RNA in the presence of insulin. Removal of insulin stabilizes the target RNA and allows protein expression (Trong et al., Biochem J., December 23, 2004).

[0189] Other non-limiting examples of expressible heterologous sequences that may be used in the compositions and methods of the present invention include sequences capable of producing proteins, including, for example, interferons (α, β, γ, ε), erythropoietin, factor VIII, coagulation factors, antibodies and fragments thereof (e.g., including single-chain, Fab, and humanized), insulin, chemokines, cytokines, growth factors, angiogenesis regulators, and apoptosis regulators, such as growth factors including, for example, bimodalin, B lymphocyte stimulating factor, interleukin-16 (IL16), thymopoietin, TRAIL, Apo-2, pre-B cell colony-enhancing factor, and endothelial differentiation-associated factor 1. (EDF-1), Endothelial Monocyte Activating Polypeptide II, Macrophage Migration Inhibitor Factor MIF, Natural Killer Cell Enhancer Factor (NKEFA), Bone Morphogenetic Protein 8 (Osteogenic Protein 2), Bone Morphogenetic Protein 6, Connective Tissue Growth Factor (CTGF), CGI-149 Protein (Neuroendocrine Differentiation Factor), Cytokine A3 (Macrophage Inflammatory Protein 1-α), Glioblastoma Cell Differentiation-Associated Protein (GBDR1), Hepatocellular Carcinoma Derivative Growth Factor, Neuromodulatory U-25 Progenitor, Any Oncogene, Oncogene, Proto-Oncogene, or Cell Regulatory Gene (which can be found at condor.bcm.tmc.edu / oncogene), Vascular Endothelial Growth Factor (VEGF), Vascular Endothelial Growth Factor B (VEGF-B), T Cell-Specific RANTES Progenitor, Thymic Dendritic Cell Derivative Factor 1; Receptors such as Activin A Receptor Type II (ACVR2), β Signal Sequence Receptor (SSR2), CD14 Monoclonal Antigen Nuclear cell LPS receptor, CD36 (type 1 collagen / platelet-reactive protein receptor)-like 2, CD44R (Hermes antigen gp90 homing receptor), G protein-coupled receptor 9, chemokine C×C receptor 4, colony-stimulating factor 2 receptor β (CSF2RB), FLT-3 receptor tyrosine kinase, transient receptor potential C precursor analog, cytotoxic lectin-like receptor B subfamily, low-density lipoprotein receptor gene, low-affinity Fcγ receptor IIC, MCP-1 receptor, monocyte chemotactic protein 1 receptor (CCR2), nuclear receptor 4 subfamily A member 1, orphan G protein-coupled receptor GPRC5D, peroxisome proliferation-activated receptor γ, pheromone-associated receptor (rat), vasopressin-activated calcium mobilization putative receptor, retinoic acid receptor, Toll-like receptor 6, transmembrane activator and CAML interacting protein (TACI), B cell mature peptide (BCMA), CSF-1 receptor, interferon (α, β, and γ) receptor 1 (IFNAR1). Pathways that can be modulated to increase antibody production include, for example, ubiquitin / proteasome; telomerase; FGFR3; and Mcd-1.

[0190] In some embodiments of this disclosure, the PIV5 composition can be used to prepare an antigen preparation for use as a vaccine. Any suitable antigen can be prepared according to this disclosure, including antigens obtained from prions, viruses, mycobacteria, protozoa (e.g., Plasmodium falciparum (malaria)), trypanosomes, bacteria (e.g., streptococci, Neisseria, etc.).

[0191] Host cells can be transfected with a single PIV5 particle containing one or more heterologous polynucleotide sequences, or with multiple PIV5 particles, each containing the same or different heterologous polynucleotide sequences. For example, multi-subunit antigens (including intracellular and cell surface multi-subunit components) can be prepared by expressing the individual subunits on separate vectors, but infecting the same host cell with all vectors, thereby allowing assembly to occur within the host cell.

[0192] Vaccines typically contain multiple antigenic components, such as those derived from different proteins and / or different epitope regions of the same protein. For example, a vaccine against a viral disease may contain one or more polypeptide sequences derived from a virus, which, when administered to a host, can elicit an immunogenic or protective response against viral attack.

[0193] As previously described, this disclosure can also be used to prepare polypeptide multimers, for example, wherein antigenic preparations composed of more than one polypeptide are produced. For example, a viral capsid can be composed of more than one polypeptide subunit. By transducing host cells with vectors carrying different viral envelope sequences, when proteins are expressed in the cells, these proteins can self-assemble into three-dimensional structures containing more than one protein subunit (e.g., in their native conformation).

[0194] In other embodiments, the expressible heterologous nucleotide sequence is derived from another virus besides PIV5. For example, the heterologous nucleotide sequence may encode influenza HA, RSV F, HIV Gag, and / or Env (from any strain). Such embodiments can be used to develop vaccines and / or vaccination methods. The examples given herein are non-limiting, as those skilled in the art will understand that nucleotide sequences from a variety of pathogens (including bacteria, parasites, etc.) may be suitable for CVB vaccine compositions and / or vaccination methods.

[0195] Examples of viruses from which vaccines can be generated according to this disclosure include, for example, coronaviruses, orthomyxoviruses, influenza A viruses (including all strains with different HA and NA proteins, such as (non-limiting examples) H1N1, H1N2, H2N2, H3N2, H7N7, and H3N8); influenza B viruses, influenza C viruses, sogoto viruses (including Dorivirus, Batken virus, SiAR 126 virus), Isaviruses (e.g., infectious salmon anemia virus), and Gram-negative bacteria (including Burkholderia melioides, Borrelia burgdorferi, Chlamydia, etc.). These include Gram-negative bacteria isolated or transmitted from all species types, including those isolated from invertebrates, vertebrates, mammals, humans, non-human primates, monkeys, pigs, dairy cows and other livestock, birds, poultry (such as turkeys, chickens, quails, and ducks), wild birds (including waterfowl and landfowl), reptiles, etc. These also include existing strains that have been altered, for example, through mutation, antigenic drift, antigenic conversion, recombination, etc., especially strains that have been enhanced in virulence and / or spread across species (e.g., human-to-human transmission).

[0196] V. Application method

[0197] A. Through vaccination

[0198] The present invention includes a method for vaccinating a subject by administering a viral expression vector, viral particle, or composition as described herein.

[0199] This disclosure provides vaccines against Gram-negative bacteria, including existing serotypes, their derivatives and recombinants, such as subtypes and recombinants capable of human-to-human transmission.

[0200] This disclosure also provides a method for producing a PIV5 composition. The host cells that can be used to produce the PIV5 composition include any mammalian or human cell line or primary cells. Non-limiting examples include, for example, 293, HT1080, Jurkat, and SupT1 cells. Other examples include CHO, 293, HeLa, Vero, L929, BHK, NIH 3T3, MRC-5, BAE-1, HEP-G2, NSO, U937, Namalwa, HL60, WEHI 231, YAC 1, U 266B1, SH-SY5Y, CHO (e.g., CHO-K1, CCL-61), and 293 (e.g., CRL-1573). Cells are cultured under conditions that enable efficient transfection and expression. These conditions, for example, include specific environments required to achieve protein production. Such an environment includes, for example, appropriate buffers, oxidants, reducing agents, pH, cofactors, temperature, ion concentration, appropriate cell age and / or stage when using cells (such as at a specific part of the cell cycle or at a specific stage of gene expression), and culture conditions (including cell culture medium, substrate, oxygen, carbon dioxide, glucose and other sugar substrates, serum, growth factors, etc.).

[0201] This disclosure also provides various treatment methods, including in vivo delivery of PIV5 to host cells. In some embodiments, PIV5 is delivered to a subject for the treatment or prevention of Gram-negative bacterial infections. In other embodiments, PIV5 is delivered to a subject for the treatment or prevention of Burkholderia melioides, Borrelia burgdorferi, or Chlamydia trachomatis infections, or to elicit an immune response in the subject against Burkholderia melioides, Borrelia burgdorferi, or Chlamydia trachomatis infections.

[0202] It is conceivable that the compositions and methods of this disclosure, when used to treat various diseases, can be used in combination with other therapeutic agents suitable for the same or similar diseases. Furthermore, two or more embodiments of this disclosure can also be co-administered to produce an additive or synergistic effect. When co-administered with a second therapeutic agent, the embodiments of this disclosure and the second therapeutic agent can be administered simultaneously or sequentially (in any order). Due to additive or synergistic effects, the appropriate therapeutically effective dose of each agent may be reduced.

[0203] As a non-limiting example, this disclosure can be combined with other therapies that block inflammation by means of (e.g., inhibiting, reducing, and / or blocking IL1, INFα / β, IL6, TNF, L13, IL23, etc.). In some embodiments, the PIV5 compositions and methods disclosed herein can be used to enhance the efficacy of vaccines against Gram-negative bacterial infections. The compositions and methods of this disclosure can be administered to a subject simultaneously with or before (e.g., 1–30 days prior) an agent (including, but not limited to, small molecules, antibodies, or cellular reagents) that induces an immune response (e.g., against infection). The compositions and methods of this disclosure can also be administered in combination with antibodies against pathogenic antigens or allergens.

[0204] The pharmaceutical compositions of the present invention are readily applicable to a variety of therapeutic or preventative applications, for example, for treating Gram-negative bacterial infections or inducing an immune response against Gram-negative bacteria in an infected subject. In various embodiments, the vaccine compositions can be used to treat or prevent infections caused by pathogens derived from immunogenic peptides displayed in PIV5-based vaccines. Therefore, the vaccine compositions of the present invention can be used in various clinical settings to treat or prevent infections caused by a variety of Gram-negative bacteria. As an example, vaccine compositions based on *Burkholderia melioides* and *Burkholderia melioides* Pal can be administered to subjects to induce an immune response to sepsis, for example, by inducing broad-spectrum neutralizing antibodies against *Burkholderia melioides* and *Burkholderia melioides*. In another example, vaccine compositions based on *Bretorius burgdorferi* OspA can be administered to subjects to induce an immune response to Lyme disease, for example, by inducing broad-spectrum neutralizing antibodies against *Bretorius burgdorferi*. In another example, a Chlamydia MOMP-based vaccine composition can be administered to a subject to induce an immune response to Chlamydia, for example, to induce the production of broad-spectrum neutralizing antibodies against Chlamydia bacteria. For subjects at risk of infection with these bacteria, the vaccine composition of the present invention can be administered to provide protection against viral infection. Therapeutic and prophylactic applications of vaccines derived from other immunogens described herein can also be carried out similarly. Depending on the specific subject and disease, the pharmaceutical compositions of the present invention can be administered to subjects via a variety of administration methods known to those skilled in the art, such as local, oral, intranasal, intramuscular, subcutaneous, intravenous, intra-articular, intra-articular, intraperitoneal, or parenteral routes. In some aspects, administration is on a mucosal surface. Vaccines can be administered using mass administration techniques, such as placing the vaccine in drinking water or spraying it into the animal's living environment. When administered by injection, the immunogenic composition or vaccine can be administered parenterally. Parenteral administration includes, for example, administration by intravenous, subcutaneous, intramuscular, or intraperitoneal injection.

[0205] 1. Administration by inhalation

[0206] In one embodiment, the disclosed PIV5 vaccine composition is formulated for intranasal administration. The intranasal composition may comprise an inhalable dry powder pharmaceutical formulation containing a therapeutic agent, wherein the therapeutic agent is present in the form of a free base or a mixture of a salt and a free base. The pharmaceutical formulations disclosed herein can be formulated for respiratory administration, such as nasal, intranasal, sinus, oral, and / or pulmonary administration. Typically, the formulation is manufactured to have an appropriate particle size suitable for the airway route or target. Therefore, the formulations disclosed herein can be formulated to have a defined particle size distribution.

[0207] For example, the particle size distribution of the salt form of the therapeutic agent for intranasal application can be between about 5 μm and about 350 μm. More specifically, the salt form of the therapeutic agent for intranasal application can have the following particle size distributions: about 5 μm to about 250 μm, about 10 μm to about 200 μm, about 15 μm to about 150 μm, about 20 μm to about 100 μm, about 38 μm to about 100 μm, about 53 μm to about 100 μm, about 53 μm to about 150 μm, or about 20 μm to about 53 μm. The salt form of the therapeutic agent in the pharmaceutical composition of the present invention for intranasal application can have a particle size distribution range of less than about 200 μm. In other embodiments, the salt form of the therapeutic agent in the pharmaceutical composition has the following particle size distributions: less than about 150 μm, less than about 100 μm, less than about 53 μm, less than about 38 μm, less than about 20 μm, less than about 10 μm, or less than about 5 μm. The salt form of the therapeutic agent in the pharmaceutical composition of the present invention for intranasal application may have the following particle size distribution ranges: greater than about 5 μm, greater than about 10 μm, greater than about 15 μm, greater than about 20 μm, greater than about 38 μm, less than about 53 μm, less than about 70 μm, greater than about 100 μm, or greater than about 150 μm.

[0208] Additionally, the salt form of the therapeutic agent in the pharmaceutical composition of the present invention for pulmonary application may have a particle size distribution range between about 1 μm and about 10 μm. In other embodiments for pulmonary application, the particle size distribution range is between about 1 μm and about 5 μm or between about 2 μm and about 5 μm. In other embodiments, the salt form of the therapeutic agent has the following average particle sizes: at least 1 μm, at least 2 μm, at least 3 μm, at least 4 μm, at least 5 μm, at least 10 μm, at least 20 μm, at least 25 μm, at least 30 μm, at least 40 μm, at least 50 μm, at least 60 μm, at least 70 μm, at least 80 μm, at least 90 μm, or at least 100 μm.

[0209] In some embodiments, the disclosed cannabinoid compositions comprise one or more cannabinoids or pharmaceutically acceptable derivatives or salts thereof, a propellant, an alcohol, and ethylene glycol and / or ethylene glycol ethers. The alcohol may be a monohydric alcohol or a polyhydric alcohol, and preferably a monohydric alcohol. The viscosity of a monohydric alcohol is lower than that of ethylene glycol or ethylene glycol ethers. Therefore, compared to compositions without monohydric alcohols, this composition is capable of forming droplets with smaller diameters. The inventors have surprisingly discovered that a specific ratio of monohydric alcohol to ethylene glycol or ethylene glycol ether yields a composition with a desirable combination of long-term stability (e.g., the composition remains single-phase for at least one week at temperatures of 2-40°C) and small droplet size.

[0210] B. Boosting vaccines

[0211] This disclosure provides the administration of a PIV5 booster vaccine in a method for inducing an immune response in human subjects, wherein the subjects have previously received a primary vaccination against a bacterial infection.

[0212] The booster vaccination method according to this disclosure includes the step of administering a vaccine composition to a subject.

[0213] The immune response induced by the vaccine composition or method disclosed herein is preferably a humoral response, particularly including a response that produces neutralizing antibodies against Gram-negative bacteria, i.e., a neutralizing antibody response.

[0214] Example

[0215] Example 1: Burkholderia (Pal) construct based on PIV5 and AVLP

[0216] To generate the PIV5-Pal (MHD11) construct, a gene fragment encoding amino acid segments 22-170 of the *Burkholderia glandis* (ATCC23344) Pal protein was synthesized, and codon usage was optimized for human cells (GenScript custom gene synthesis service). The Pal protein was inserted between the PIV5 SH and HN linker sites to generate MHD11 (PIV5-Pal). For the PIV5-NTTM-Pal (MHD39) construct, the N-terminus and transmembrane domain of the PIV5 HN glycoprotein were directly incorporated into the N-terminus of the Pal protein. Figure 1 Modified Pal cDNA was cloned between the SH and HN genes of PIV5 in plasmid ZL185, which contains the entire genome of the PIV5 virus. MHD11 and MHD39 viruses were rescued by co-transfecting MHD11 and MHD39 plasmids with pPIV5-NP, pPIV5-P, pPIV5-L, and pT7-polymerase plasmids encoding NP, P, and L proteins, as well as T7 RNA polymerase, into BHK21 cells.

[0217] To test the expression of Pal protein in MHD11 and MHD39 infected cells, cell lysates and supernatants were subjected to SDS-PAGE at 4-20%. Acrylamide gel Separate from the sample and then transfer it to Amersham Hybond LFP. PVDF On the membrane. The membrane was incubated with a mixture of mouse anti-Pal mAb and mouse anti-PIV5-NP antibody, and then bound to Cy3. goat Anti-mouse IgG incubation ( Figure 2 Compared to Pal alone, the addition of the PIV5 HN NT+TM domain increases the expression of Pal protein in cell lysates, and as shown in the culture supernatant, Pal protein can also be incorporated into viral particles.

[0218] To generate the AVLP-Pal (MHD30) construct, the EGFP gene in the pAE31 plasmid containing the AVLP genome was replaced with the same gene fragment as MHD11. For the AVLP-NTTM-Pal (GOB10) construct, the N-terminus and transmembrane domain of the PIV5 HN were inserted into the N-terminus of the Pal protein. Stable BHK21 cell lines containing the MHD30 and GOB10 constructs were formed by co-transfecting the MHD30 and GOB10 plasmids with pPIV5-NP, pPIV5-P, pPIV5-L, and pT7-polymerase plasmids encoding NP, P, and L proteins and T7 RNA polymerase under selected conditions. After cell line formation, AVPPS were created by co-transfecting the PIV5-F, PIV5-HN, and PIV5-M plasmids and collecting the culture medium. Descriptions of these constructs can be found in [link to relevant documentation]. Figure 3A middle.

[0219] The immunogenicity of the MHD30 and GOB10 constructs was measured in a mouse model. Figure 3B In short, BALB / c mice aged 6 to 8 weeks were treated with a dose of 3.8 x 10⁻⁶. 5 Mice were immunized intranasally with PBS, MHD30, or GOB10. Blood samples were collected from mice 14 days post-immunization. A booster immunization was administered intranasally 21 days post-immunization with the same dose as the initial immunization. Seven days post-booster, mice were euthanized humanely, and blood was collected for serological analysis.

[0220] ELISA was used to determine the levels of anti-Pal IgG antibodies in serum 14 days after primary immunization and 7 days after booster immunization (day 28). Briefly, Immulon® 2HB 96-well microplates were coated with 1 μg / ml purified Pal protein PBS solution. Serum samples were serially diluted twofold in blocking buffer and added to the plates. After washing the serum, secondary antibody (horseradish peroxidase-labeled goat anti-mouse IgG) was added to each well. The endpoint titer was defined as the highest serum dilution with an OD value exceeding two standard deviations of the mean OD value of untreated serum. Figure 3B The results showed that the addition of PIV5 HN NT+™ significantly improved the immunogenicity induced by Pal protein at 14 days after primary immunization and 7 days after booster immunization. MHD30 (AVLP-Pal) was not immunogenic in the mouse model.

[0221] A second mouse immunogenicity study was conducted to compare the MHD30 and GOB10 constructs with the MHD11 and MHD39 vaccines. In short, 6- to 8-week-old BALB / c mice were administered a dose of 3.8 x 10⁻⁶. 5 Mice were intranasally immunized with AP or PFU in PBS, MHD30, GOB10, MHD11, or MHD39. Blood was collected from mice 14 days post-immunization. A booster immunization was administered intranasally 21 days post-immunization with the same dose as the initial immunization. Seven days post-boost immunization, mice were humanely euthanized, and blood was collected for serological analysis.

[0222] ELISA was used to determine the levels of anti-Pal IgG antibodies in serum 14 days after primary immunization and 7 days after booster immunization (day 28). Briefly, Immulon® 2HB 96-well microplates were coated with 1 μg / ml purified Pal protein PBS solution. Serum samples were serially diluted twofold in blocking buffer and added to the plates. After washing the serum, secondary antibody (horseradish peroxidase-labeled goat anti-mouse IgG) was added to each well. The endpoint titer was defined as the highest serum dilution with an OD value exceeding two standard deviations of the mean OD value of untreated serum. Figure 3C The results showed that adding PIV5 HN NT+™ to the replicative PIV5 backbone and the replication-deficient AVLP platform significantly improved Pal protein-induced immunogenicity at 14 days after primary immunization and 7 days after booster immunization. MHD30 (AVLP-Pal) was not immunogenic in the mouse model.

[0223] Example 2. PIV5-based Lyme disease (OspA) construct

[0224] OspA is a lipoprotein present in the periplasm and liquid outer membrane, and is present in Borrelia burgdorferi when it is in the midgut of a tick (Li, H., et al., Proceedings of the National Academy of Sciences 94, 3584-3589 (1997)).

[0225] To generate the MHD22 plasmid construct, the OspA BPBPk gene, codon-optimized for human use, was incorporated into the pCAGGS expression plasmid. For the MHD24 plasmid, the N-terminus and transmembrane domain (TM) sequence of the PIV5 HN protein were directly added upstream of the OspA antigen sequence. This was achieved by adding NTTM-OspA... BPBPk Or NTTM-OspA B31 Sequences were introduced between the SH and HN genes of the PIV5 genome, generating the MHD35 and MHD36 constructs. The OspA BPBPk sequence was created by replacing the amino acid sequences of Borrelia burgdorferi B31165-189 and 219-273 with the corresponding sequences of Borrelia auriculata strains to avoid any cross-reaction with autoimmune epitopes (del Rio, B. et al., Clinical and vaccine immunology: CVI 15, 1429-1435 (2008)). The CVL106 construct was created by introducing the NTTM-OspA... BPBPk The CVL116 construct was generated by introducing the gene sequence between the SH and HN genes in the CPIPIV5 genome, while the CVL116 construct was generated by introducing the same antigen into the CVBPIV5 genome to replace the SH gene. All viruses were rescued by co-transfecting the MHD35, MHD36, CVL106, and CVL116 plasmids with pPIV5-NP, pPIV5-P, pPIV5-L, and pT7-polymerase plasmids encoding NP, P, and L proteins and T7 RNA polymerase into cells. Descriptions of these constructs are included in [the relevant section]. Figures 4A-4B middle.

[0226] MHD22 and MHD24 pCAGGS constructs, along with an MHD control, were transfected into BHK cells for Western blot analysis. In short, cell lysates were subjected to SDS-PAGE at 4-20% concentration. Acrylamide gel Separate from the sample and then transfer it to Amersham Hybond LFP. PVDF On the membrane. The membrane was incubated with mouse anti-OspA antibody and mouse anti-actin antibody, and then bound to Cy3. goat Anti-mouse IgG incubation ( Figures 5A-5BThese results indicate that, compared to OspA alone... BPBPk In comparison, the addition of PIV5 HN NT+™ increased OspA in the cell lysate. BPBPk Protein expression.

[0227] OspA is determined using the pCAGGS construct via IFA. BPBPk Protein expression on the cell surface. Cells were transfected with MHD22 and MHD24 constructs, and protein expression was detected using mouse anti-OspA antibody and anti-mouse Alexa Fluor 488 secondary antibody. Figures 6A-6C The results showed that, compared with MHD22, OspA in cells transfected with the MHD24 construct was significantly lower. BPBPk Increased protein expression indicates that the addition of PIV5 HN NT+™ can increase OspA. BPBPk Expression on the cell surface.

[0228] MHD35 (PIV5-A) was analyzed by Western blot assay. BPBPk ) and MHD36 (PIV5-A B31 OspA protein expression in the vaccine construct. In short, cell lysates were subjected to SDS-PAGE at 4-20%. Acrylamide gel Separate from the sample and then transfer it to Amersham Hybond LFP. PVDF On the membrane. The membrane was incubated with mouse anti-OspA antibody and mouse anti-PIV5 NP antibody, and then bound to Cy3. goat Anti-mouse IgG incubation ( Figure 7 These results indicate that both MHD35 and MHD36 viruses express the OspA protein.

[0229] To evaluate the PIV5-A BPBPk PIV5-A B31 and SC OspA B31 Vaccine-induced humoral IgG immune responses were assessed, and all blood samples collected prior to tick challenge were used to determine anti-OspA antibodies (1:100) by ELISA. Compared to controls, sera from mice vaccinated with OspA (IN and SC) typically showed anti-OspA IgG antibody ODs collected before booster immunization (D17) and after booster immunization (>D86). 450 > 3.5. To quantify the endpoint titer of anti-OspA IgG, serum samples collected on day 17 (before booster immunization) and at 3, 6, and 12 months after the initial immunization in Study 3 (15-month challenge) were used. Figure 8 Dilute to 10 on ELISA. 2 -10 6 From SC rOspAB31 +Alum, PIV5-A BPBPk and PIV5-A B31 Mice in the vaccine group reached peak serum IgG endpoint titers (EPT) 3 months after primary immunization, with geometric mean values ​​of 5.5, 5.5, and 5.4 log₂O₅, respectively. 10 These values ​​were significantly higher than serum IgG EPT on day 17 after the initial immunization, indicating that the second dose of vaccine had a enhancing effect. In the subsequent 9-month trial, anti-OspA... B31 Serum IgG antibody levels decreased, including SC rOspA. B31 The mice in the +Alum group showed the largest decrease, at 1.7 log... 10 EPT, while PIV5-A BPBPk and PIV5-A B31 The decrease in mice in the vaccine group was 0.7 log. 10 EPT. Throughout the study, mice in the SC PBS+Alum and IN PIV5 control groups had very low levels of cross-reactive IgG antibodies at some time point, but the peak titers of these control groups were 3 log lower than the peak titers of all vaccine groups. 10 These results suggest that immunization with intranasal PIV5 vaccine provides longer-lasting protection compared to subcutaneously administered protein vaccines.

[0230] Similar to Study 3, mice in Study 2 (9 months, no attack) received either two subcutaneous doses of alum alone (SC PBS + Alum) or alum plus 20 μg rOspA protein (SC rOspA) B31 +Alum); or two intranasal doses of 10 6 PFU wild-type PIV5 (IN PIV5), PIV5-A BPBPk Or PIV5-A B31 Study 1 included the same groups except for the SC PBS+Alum group. In this study, blood was collected on day 117 for a neutralization assay of cultured Borrelia burgdorferi. Figure 9A The mice were then challenged (4 months prior to challenge). Due to insufficient blood volume from each mouse, serum from each group was pooled for neutralization assays. Figure 9A In lifespan studies (Studies 2 and 3), several additional groups of mice were vaccinated for neutralization assays, thus at day 270 (Study 2, 9 months, no attack, Figure 9B ) and day 533 (Study 3, 18 months, no attack, Figure 9CSufficient serum was collected at the time of euthanasia of each mouse. Total active bacteria were measured on days 0, 2, 5, and 7 after Study 1, and on days 0, 3, and 6 after Studies 2 and 3. Serum was compared with SC rOspA at 4 months post-primary booster immunization (Study 1). B31 +Alum, PIV5-A BPBPk and PIV5-A B31 The levels of active bacteria in cultures co-incubated with serum from the vaccine group decreased by 1.6, 0.6, and 1.0 log on day 2 compared to day 0, respectively. 10 ( Figure 9A These values ​​decreased further on day 5, until all vaccine groups had reached 0 by day 7 after neutralization. Figure 9A In contrast, the BbBSK and IN PIV5 control groups showed an increase in active bacterial counts on days 2 and 5 post-neutralization, peaking on day 5 post-inoculation, with geometric mean active Borrelia burgdorferi counts of 7.6 and 7.8 log per milliliter of culture, respectively. 10。 On day 7 after neutralization, these values ​​decreased slightly, with 7.1 and 6.2 log [values] of active Borrelia burgdorferi per milliliter of culture, respectively. 10 Nine months after the initial booster immunization (Study 2), the SC rOspA levels in the vaccine group were [data missing]. B31 +Alum, PIV5-A BPBPk and PIV5-A B31 The number of active bacteria in cultures on day 3 or 6 post-inoculation was significantly reduced compared to day 0, with almost all samples in each group reaching 0 on day 3 after neutralization. Figure 9B On the other hand, compared to day 0, samples from the Bb BSK, SC PBS+Alum, and IN PIV5 control groups showed increased levels of active bacteria in cultures on day 6 after neutralization. On average, the level of active Borrelia burgdorferi per milliliter of culture in the control groups increased by 0.2% on day 6 after neutralization compared to day 0. log10。 At 18 months following the initial-boost immunization (Study 3), data continued to show that, compared to day 0, SC rOspA B31 +Alum, PIV5-A BPBPk and PIV5-A B31 In the vaccine group, the levels of active Borrelia burgdorferi per milliliter of culture were significantly reduced in samples taken on days 3 and 6 after neutralization, with the largest differences observed on day 6, averaging decreases of 0.8, 1.5, and 2.0, respectively. log10 ( Figure 9CIn contrast, the number of active Borrelia burgdorferi per milliliter of culture on day 6 after neutralization in the Bb BSK and IN PIV5 control groups either increased or remained unchanged compared to day 0, with 0.2 log [unclear] observed in the Bb BSK samples. 10 These results indicate that intranasal PIV5 vaccines carrying OspA sequences from different broad Borrelia burgdorferi gene species can generate a robust neutralizing IgG immune response that can last up to 18 months, and these responses are more pronounced than those observed with recombinant protein vaccines.

[0231] Mice from three studies were challenged with ticks infected with various strains of Borrelia burgdorferi at 4 months (Study 1), 9 months (Study 2), or 15 months (Study 3) after primary immunization. To assess the spread of Borrelia burgdorferi to target tissues, the FlaB load of Borrelia burgdorferi in bladder, joint, and heart samples was detected by qPCR. Figures 10A-10C In addition, heart and bladder tissues were cultured in BSK-H medium to assess the motility / viability of Borrelia burgdorferi under a dark-field microscope and confirmed by FlaB PCR of the cultures. Figure 10C-10F In Study 1, using SCrOspA... B31 IN PIV5-A BPBPk and IN PIV5-A B31 Challenge was performed 4 months after the initial-boost immunization, and results showed no Borrelia burgdorferi FlaB DNA in the heart, bladder, and joints, nor any live Borrelia burgdorferi in heart cultures, in contrast to controls receiving intranasal (IN) PIV5 or subcutaneous (SC) PBS + alum. In Study 2, when using INPIV5-A... BPBPk and IN PIV5-A B31 Nine months after the initial-boost immunization, a challenge was performed, and results showed no Borrelia burgdorferi FlaB DNA in the heart, bladder, and joints, and no live Borrelia burgdorferi in bladder cultures. This was consistent with the results of SC rOspA administration. B31 One-quarter of the mice were compared with all controls who received intranasal (IN) PIV5 or subcutaneous (SC) PBS+alum. In Study 3, mice using IN PIV5-A... B31 Challenge was performed 15 months after the initial-boost immunization, and results showed no Borrelia burgdorferi FlaB DNA in the heart, bladder, and joints, nor were there any live Borrelia burgdorferi in bladder cultures, which is consistent with 1 / 3 IN PIV5-A. BPBPk Accept SC rOspA B31Three-thirds of the mice were compared with all controls that received intranasal (IN) PIV5 or s...

Claims

1. A recombinant vaccine comprising a PIV5-based antigen expression vector, said vector comprising: PIV5-based viral expression vectors; and Genes that express antigens in mammalian cells The gene expressing the antigen is modified to increase the expression of the antigen on the cell surface (AOS); The gene expressing the modified antigen is placed in the PIV5 genome; and The antigens mentioned therein are viral antigens or bacterial antigens.

2. The vaccine of claim 1, wherein the PIV5-based antigen expression vector has a heterologous nucleic acid sequence with at least 98% sequence identity to SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12.

3. The vaccine of claim 1, wherein the PIV5-based viral expression vector comprises a PIV5-AVLP, PIV5-W3A, PIV5-CPI, or PIV5-CVB backbone.

4. The vaccine of claim 1, wherein the modified antigen gene is modified by replacing the signal peptide of the gene with the N-terminal domain (NT) of the PIV5 HN protein or the N-terminal domain of the PIV5 HN protein plus a transmembrane domain (NTTM).

5. The vaccine of claim 1, wherein the modified antigen gene is modified by directly adding only the N-terminal domain (NT) of the PIV5 HN protein upstream of the antigen or by adding the N-terminal domain of the PIV5 HN protein plus a transmembrane domain (NTTM) sequence without removing the signal peptide.

6. The vaccine of claim 1, wherein the modified antigen gene is positioned between the SH and HN genes or the HN and L genes of the PIV5 genome.

7. The vaccine of claim 1, wherein the modified antigen gene replaces the SH gene of the PIV5 genome.

8. The vaccine of claim 1, wherein the modified antigen gene is expressed on the surface of the mammalian cells.

9. The vaccine of claim 1, wherein the bacterial antigen is an antigen of Gram-negative bacteria selected from the group consisting of Burkholderia melioides, Borrelia burgdorferi, and Chlamydia trachomatis; and wherein the viral antigen is an antigen of viruses selected from the group consisting of coronaviruses, respiratory syncytial virus (RSV), influenza A virus, influenza B virus, influenza C virus, orthomyxovirus, Sogoto virus, and Isa virus.

10. The vaccine of claim 9, wherein the gene expressing the bacterial antigen is modified by replacing the N-terminus of the Burkholderia melioides Pal protein with the N-terminus and transmembrane domain of the PIV5 HN glycoprotein.

11. The vaccine of claim 9, wherein the gene expressing the bacterial antigen is modified by placing the N-terminus and transmembrane domain of the PIV5 HN glycoprotein directly upstream of the Borrelia burgdorferi OspA BPBPk protein.

12. The vaccine of claim 9, wherein the gene expressing the bacterial antigen is modified by replacing the signal peptide of the Chlamydia trachomatis major outer membrane protein (MOMP) with the N-terminus of the PIV5 HN glycoprotein alone or with the N-terminus and transmembrane domain, or by placing the N-terminus and transmembrane domain of the PIV5 HN glycoprotein directly upstream of the antigen without removing the signal peptide.

13. A PIV5-based antigen vaccine composition comprising... PIV5-based viral expression vectors; and Genes that express bacterial antigens in mammalian cells. The antigens mentioned therein are viral antigens or bacterial antigens; The PIV5-based viral expression vector comprises a PIV5-AVLP, PIV5-W3A, PIV5-CPI, or PIV5-CVB backbone. The gene expressing the antigen is modified in the following ways: by replacing the signal peptide of the gene with the N-terminal domain (NT) of the PIV5 HN protein or the N-terminal domain plus transmembrane domain (NTTM) of the PIV5 HN protein; or by directly adding only the N-terminal domain (NT) of the PIV5 HN protein or adding the N-terminal domain plus transmembrane domain (NTTM) sequence upstream of the antigen without removing the signal peptide; and The gene expressing the modified antigen is placed between the SH and HN genes or between the HN and L genes in the PIV5 genome, or by replacing the SH gene.

14. The composition of claim 13, wherein the PIV5-based antigen vector has a nucleic acid sequence having at least 98% sequence identity with SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12.

15. The composition of claim 13, wherein the composition is a vaccine against bacterial or viral infections.

16. The composition of claim 13, wherein the bacterial infection is caused by Gram-negative bacteria selected from the group consisting of Burkholderia melioides, Borrelia burgdorferi, and Chlamydia trachomatis; and the viral infection is caused by viruses selected from the group consisting of coronaviruses, respiratory syncytial virus (RSV), influenza A virus, influenza B virus, influenza C virus, orthomyxovirus, Sogoto virus, and Isa virus.

17. A method for inducing an immune response in a subject in need, the method comprising administering to the subject the PIV5-based antigen vaccine of claim 1, wherein the immune response comprises a humoral immune response and / or a cellular immune response, and wherein the subject is susceptible to bacterial or viral infection.

18. The method of claim 17, wherein the subject has previously been vaccinated against the bacterial or viral infection, the method comprising administering the composition to the subject.

19. The method of claim 17, wherein the bacterial infection is caused by Gram-negative bacteria selected from the group consisting of Burkholderia melioides, Borrelia burgdorferi, and Chlamydia trachomatis; and the viral infection is caused by viruses selected from the group consisting of coronaviruses, respiratory syncytial virus (RSV), influenza A virus, influenza B virus, influenza C virus, orthomyxovirus, Sogoto virus, and Isa virus.

20. The method of claim 17, wherein the composition is administered intranasally, intramuscularly, topically, parenterally, or orally.

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