Engineering broadly reactive coronavirus vaccines and related designs and uses

Abstract

To provide a vaccine for preventing β-CoV infection.SOLUTION: The present invention provides a vaccine comprising: a viral vector containing a SARSβ-CoV DNA sequence which encodes at least part of S protein for β-CoV, wherein the vector comprises a fully deleted adenovirus vector free of all endogenous genes, the sequence comprises a consensus sequence derived from a homology group containing a plurality of SARS-2 β-CoV and having at least 65% of a sequence identity to each member, and the variable domain of the consensus sequence is deleted from the consensus sequence and replaced with a linker peptide designed to bridge the gap; and at least one packaging plasmid based on an adenovirus selected from the group consisting of the serotypes Ad2, Ad5, Ad6, and Ad36, and combinations thereof, wherein the at least one packaging plasmid comprises a single left ITR or right ITR that confers replication deficiency.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to, and is not a provisional application, Ser. No. 63 / 012,360, filed April 20, 2020, which is incorporated herein by reference in its entirety.

[0002] The present disclosure relates to a vaccine platform for developing coronavirus vaccines, more particularly vaccines for protecting mammals against infection from β-coronaviruses. In another embodiment, the present disclosure relates to a method for developing coronavirus vaccines using the identified group of gene sequences.

[0003] Submission of sequence listing The contents of the electronic submission of the Sequence Listing, a text file entitled "Sequence_Listing", created on April 20, 2021, and having a size of 54, is hereby incorporated by reference in its entirety. [Background technology]

[0004] Coronaviruses (CoVs) are classified into four genera: alpha-, beta-, gamma- and delta-coronaviruses. β-CoVs are enveloped, positive-stranded RNA viruses that can infect mammals, typically bats and rodents, although many β-CoVs are known to infect humans as well. Infections with CoVs in humans and animals generally cause a short-lived, mild to moderate upper respiratory tract disease. Exceptions are Severe Acute Respiratory Syndrome (SARS-1), Middle East Respiratory Syndrome (MERS), and SARS-CoV-2 (SARS-2), also known as COVID-19, which originated in Wuhan, characterized by severe and often fatal symptoms. The first cases of SARS-2 infection were seen in December 2019. As of April 16, 2020, in the United States alone, there were an estimated 632,000 cases and an estimated 31,000 deaths reported by the Centers for Disease Control and Prevention (CDC), resulting in a case fatality rate of 4.9%. SARS-2 is highly infectious in humans. On 30 January 2020, the World Health Organization (WHO) declared the global SARS-2 pandemic a global health emergency.

[0005] No specific treatment for SARS-2 is available, but is under investigation. The best approach to prevent further spread of the disease is the development of a specific vaccine. Herd immunity against SARS-2 is better achieved with immunization by a benign vaccine rather than natural infection with the active SARS-2 virus. One explanation for the low level of immune response seen in recovering patients could be a function of exhausting immune suppression by SARS-2. However, animal studies with traditional vaccines that use inactivated forms of the virus suggest that inactivated virus vaccines may be particularly susceptible to the induction of antibody-dependent enhancement (ADE) of disease. In the case of these vaccinations, Th2-type disease enhancement may be driven by anti-nucleocapsid (NP) responses. It would be desirable to develop a SARS-2 vaccine that does not stimulate ADE in vaccine recipients.

[0006] Although social distancing has successfully suppressed active transmission of SARS-2, reopening of society is expected to lead to a short-term surge in infections as well as possible seasonal outbreaks. Some regions have already seen a surge in infections caused by mutated SARS-2 viruses. The overall mutation rate of SARS-related β-CoV (SARSr) has been calculated as low as 0.1 mutations per generation. Despite the emergence of recent mutations, the SARS-2 virus appears to be stable as well. It would be desirable for any SARS-2 vaccine to also provide protection against short-term variants.

[0007] Numerous animal and clinical trials with the related SARS- and MERS-CoVs suggest that it may be possible to produce effective vaccines against the more common -CoV infections. SARS-2 (COVID-19) is a third deadly β-CoV that has jumped from an animal host to humans. Considering that 1,800 SARSr have already been identified in animals, some of which may eventually infect humans, it would be desirable to also create a group-specific SARSr vaccine to avoid future pandemics.

[0008] The use of viral vectors, including adenovirus-based viral vectors, in vaccines is known. Such "ad vectors" have repeatedly demonstrated higher and more sustained immunogenicity compared to other vaccine systems. One problem with using ad vectors in vaccination programs is the strong immune response elicited against the adenovirus itself, as opposed to the target virus. To avoid these strong anti-adenovirus responses, (fd)ad vectors were developed that completely deleted all endogenous adenovirus genes. The packaging information of the fd adenovirus genome was initially delivered together with a second viral construct-hybrid baculovirus-adenovirus or helper virus. Unfortunately, this resulted in the contamination of the replication elements of the ad vector or helper virus. It is desirable to develop an ad vector vaccine system that avoids these problems with existing ad vector vaccines. Summary of the Invention

[0009] In one embodiment, the present disclosure provides a vaccine for preventing β-CoV infection. According to an embodiment of the present disclosure, the vaccine for preventing β-CoV infection comprises at least one viral vector comprising a β-CoV DNA sequence encoding the S protein of β-CoV.

[0010] In one embodiment, the vector is an adenoviral vector. In another embodiment, the vector is a fully deleted adenoviral vector that does not contain all endogenous genes. In yet another embodiment, the β-CoV DNA sequence is a SARS-2β-CoV DNA sequence. In a further embodiment, the SARS-2β-CoV DNA sequence is the entire sequence encoding the S protein. In yet a further embodiment, the SARS-2β-CoV DNA sequence is a partial sequence encoding the S protein. In another embodiment, the SARS-2β-CoV DNA sequence is a partial sequence encoding the S protein from which the receptor binding domain has been removed. In yet another embodiment, the SARS-2β-CoV DNA sequence is a partial sequence encoding the S protein in which the receptor binding domain sequence has been replaced by DNA encoding a peptide linker.

[0011] In one embodiment, the vaccine further comprises a packaging plasmid based on an adenovirus selected from the group consisting of serotypes Ad2, Ad5, Ad6 and Ad35, and combinations thereof. In a further embodiment, the at least one viral vector is contained in a packaging cell. In yet another embodiment, the packaging cell is encapsidated into a capsid selected from the group consisting of serotypes Ad2, Ad5, Ad6 and Ad35, and combinations thereof.

[0012] In one embodiment, the β-CoV DNA sequence is a SARS-2 β-CoV DNA sequence, and the viral vector comprises at least a second β-CoV DNA sequence derived from a SARSr virus, which second β-CoV DNA sequence encodes the S protein of the SARSr virus.

[0013] In one embodiment, the present disclosure provides a vaccine for preventing SARS-2 infection. According to an embodiment of the present disclosure, the vaccine for preventing SARS-2 infection includes at least one viral vector comprising a SARS-2 β-CoV DNA sequence encoding the S protein of SARS-2 β-CoV and at least one packaging plasmid based on an adenovirus selected from the group consisting of serotypes Ad2, Ad5, Ad6 and Ad36 and combinations thereof, wherein the at least one viral vector and the at least one packaging plasmid are contained in a packaging cell, and the packaging cell is encapsidated into a capsid selected from the group consisting of serotypes Ad2, Ad5, Ad6 and Ad35 and combinations thereof.

[0014] In one embodiment, the SARS-2 β-CoV DNA sequence encodes a partial S protein of the SARS-2 virus.

[0015] In one embodiment, the present disclosure provides a vaccine for preventing β-CoV infection. According to an embodiment of the present disclosure, the vaccine for preventing β-CoV infection comprises at least one β-CoV RNA sequence encoding the S protein of β-CoV.

[0016] In one embodiment, the RNA is mRNA. In a further embodiment, the β-CoV RNA sequence is a SARS-2β-RNA sequence. In yet a further embodiment, the SARS-2β-CoV RNA sequence is the entire sequence encoding the S protein. In yet another embodiment, the SARS-2β-CoV RNA sequence is a partial sequence encoding the S protein. In yet another embodiment, the SARS-2β-CoV RNA sequence is a partial sequence encoding the S protein from which the receptor binding domain has been removed. In a further embodiment, the SARS-2β-CoV RNA sequence is a partial sequence encoding the S protein, with the receptor binding domain sequence replaced by an RNA encoding a peptide linker.

[0017] In one embodiment, the vaccine further comprises an expression vector that delivers the genetic information of β-CoV RNA, hi another embodiment, the expression vector is an engineered viral vector.

[0018] In one embodiment, the present disclosure provides a vaccine for preventing β-CoV infection. According to an embodiment of the present disclosure, the vaccine for preventing β-CoV infection comprises at least one viral vector comprising a β-CoV protein sequence encoding the S protein of β-CoV.

[0019] In one embodiment, the β-CoV RNA sequence is a SARS-2β-CoV protein sequence. In another embodiment, the SARS-2β-CoV protein sequence is the entire sequence encoding the S protein. In yet a further embodiment, the SARS-2β-CoV protein sequence is a partial sequence encoding the S protein. In yet another embodiment, the SARS-2β-CoV protein sequence is a partial S protein sequence in which the receptor binding domain has been removed. In yet another embodiment, the SARS-2β-CoV protein sequence is a partial S protein sequence in which the receptor binding domain sequence has been replaced by a peptide linker.

[0020] In one embodiment, the present disclosure provides a method of vaccinating a mammalian subject against infection from at least one group of β-CoVs. According to an embodiment of the present disclosure, the method of vaccinating a mammalian subject against infection from at least one group of β-CoVs includes classifying a broad group of β-CoVs into homology groups based on similarities in the β-CoV RNA sequences encoding their S proteins, identifying at least one consensus sequence for each homology group that has greater than 60% sequence identity to all other members of the homology group, and preparing a viral vector that includes at least a portion of the consensus sequence from at least one homology group.

[0021] In one embodiment, the consensus sequence is selected from the group consisting of a DNA sequence, an RNA sequence, a protein sequence, and combinations thereof.

[0022] In one embodiment, the step of preparing a viral vector comprises including at least a portion of a consensus sequence from two or more homology groups.

[0023] In one embodiment, the method further comprises injecting the vaccine into a mammalian subject.

[0024] In one embodiment, the present disclosure provides a method of vaccinating a mammalian subject against infection from at least one group of β-CoVs. According to an embodiment of the present disclosure, the method of vaccinating a mammalian subject against infection from at least one group of β-CoVs includes classifying a broad group of β-CoVs into homology groups based on similarity in the DNA, RNA or protein sequences of the β-CoVs encoding their S proteins, identifying at least a portion of a β-CoV protein sequence for each homology group that has greater than 60% sequence identity to all other members of the homology group, and preparing a DNA, RNA or protein vaccine that includes at least a portion of a β-CoV protein sequence from at least one homology group.

[0025] In another embodiment, the method further comprises injecting the vaccine into a mammalian subject. [Brief description of the drawings]

[0026] [Figure 1] FIG. 1 is a schematic diagram showing functional portions of the SARS-2β-CoV RNA segment encoding the S protein according to an embodiment of the present disclosure, along with the portions that are most variable and those that elicit the greatest immune response. [Diagram 2] FIG. 1 illustrates components of a vaccine according to an embodiment of the present disclosure. [Diagram 3] Figure 3 shows the activity of avian influenza vaccines using the viral vectors of the present disclosure. Specifically, Figure 3A shows the survival rate of the subject group, Figure 3B shows the body weight of the subject group, Figure 3C shows the serum antibody titer, and Figure 3D shows the lung virus titer. [Figure 4] Demonstrating the activity of MERS-CoV vaccines. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] Before describing any embodiment of the present disclosure in detail, it should be understood that the present disclosure is not limited in its application to the details of the construction and arrangement of the components set forth in the following description or illustrated in the drawings. The present disclosure is capable of other embodiments and can be practiced or carried out in various ways. It is also understood that the phraseology and terminology used herein are for the purpose of description and should not be considered as limiting. The use of "including", "comprising" or "having" and variations thereof herein means to include the items recited thereafter and equivalents thereof as well as additional items. The use of "including essentially" and "consisting essentially of" and variations thereof herein means to cover the items recited thereafter as well as equivalents and additional items that provide such equivalents and additional items so as not to substantially alter the properties, use or manufacture of the whole. The use of "consisting" and variations thereof herein means to include only the items recited thereafter and those items only.

[0028] With reference to the drawings, like numbers refer to like elements throughout. Terms such as first, second, etc. may be used herein to describe various elements, components, regions, and / or portions, but it will be understood that these elements, components, regions, and / or portions should not be limited by these terms. These terms are used only to distinguish one element, component, region, and / or portion from another element, component, region, and / or portion. Thus, a first element, component, region, or portion can be referred to as a second element, component, region, or portion without departing from the present disclosure.

[0029] Numerical ranges in this disclosure are approximations and may include values ​​outside the range unless otherwise indicated. Numerical ranges include all values ​​from and including the lower and higher values ​​in increments of one unit (unless specifically stated otherwise), provided that there is a separation of at least two units between any lower value and any higher value. As an example, if a composition, physical or other property, such as the amount of an element by weight, is 10-100, all individual values ​​such as 10, 11, 12, etc., and subranges such as 10-44, 55-70, 97-100, etc., are intended to be explicitly recited. Ranges including explicit values ​​(e.g., ranges 1, or 2, or 3-5, or 6, or 7) include any subranges between any two explicit values ​​(e.g., the range 1-7 above includes the subranges 1-2, 2-6, 5-7, 3-7, 5-6, etc.). For ranges containing values ​​less than one or containing fractional ranges greater than one (e.g., 1.1, 1.5, etc.), one unit is considered to be 0.0001, 0.001, 0.01 or 0.1, as appropriate. In ranges containing single digit numbers less than ten (e.g., 1 to 5), one unit is typically considered to be 0.1. These are only examples of what is specifically intended, and all possible combinations of numerical values ​​between the lowest and highest values ​​recited should be considered to be expressly set forth in this disclosure.

[0030] Spatial terms such as "beneath," "below," "lower," "above," "upper," and the like may be used herein for ease of description to describe the relationship of one element or feature to another element or feature as shown in the figures. It will be understood that spatially relative terms are intended to encompass different orientations depending on the orientation in use or illustration. For example, if a device in a figure is inverted, an element described as being "below" or "below" another element or feature would be oriented "above" the other element or feature. Thus, the exemplary term "below" can encompass both an orientation of above and below. The device may be otherwise oriented (rotated 90 degrees or to other orientations) and the spatially relative descriptors used herein would be interpreted accordingly.

[0031] As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items. For example, when used in a phrase such as "A and / or B," the term "and / or" is intended to include both A and B, A or B, A (alone), and B (alone). Similarly, the term "and / or" when used in a phrase such as "A, B, and / or C" is intended to encompass each of the following embodiments: A, B, and C, A, B, or C, A or C, A or B, B or C, A and C, A and B, B and C, A (alone), B (alone), and C (alone).

[0032] In one embodiment, the present disclosure provides a method for preparing a vaccine for preventing at least one β-CoV infection in a subject, particularly a mammalian subject, more particularly a human subject.

[0033] Identification of β-CoV According to an embodiment of the present disclosure, a method for preparing a vaccine for preventing at least one β-CoV infection in a subject, particularly a mammalian subject, more particularly a human subject, comprises identifying at least one β-CoV from an animal host, particularly a mammalian host. In particular, in one embodiment, a method for preparing a vaccine for preventing at least one β-CoV infection in a subject, particularly a mammalian subject, more particularly a human subject, comprises identifying at least one β-CoV from a mammalian host selected from the group consisting of bat, rat, human, and combinations thereof. In one embodiment, the at least one β-CoV comprises at least one SARSr. In another embodiment, the at least one β-CoV comprises at least one SARS-2 β-CoV.

[0034] Identification of homology groups According to an embodiment of the present disclosure, a method for preparing a vaccine for preventing at least one β-CoV infection in a subject, particularly a mammalian subject, more particularly a human subject, comprises classifying identified β-CoVs, e.g., β-CoVs identified from an animal host, into homology groups based on gene sequence similarity and preparing at least one consensus sequence for each homology group. The homology groups can be based on similarity of the entire β-CoV gene sequence, multiple parts of the β-CoV gene sequence, or a single part of the β-CoV gene sequence. The gene sequence is selected from the group consisting of a DNA sequence, an RNA sequence, a protein sequence, and combinations thereof. It will be understood that if a single β-CoV is identified, it is the only member of a single homology group.

[0035] In certain embodiments, the β-CoV comprises multiple SARSr, and the multiple SARSr are classified into one, or at least two, or at least three, or at least four, or at least five homology groups. In one embodiment, the homology groups are based on at least a portion, or at least two or more portions, or all of the gene sequences associated with the spike protein, the SARS receptor binding domain (RBD), the envelope protein, the nucleoprotein, and combinations thereof.

[0036] In a further embodiment, at least one SARS-2β-CoV is identified and classified into at least one homology group.

[0037] In one embodiment, within each homology group, the gene sequence has 60% or more, or 65% or more, or 70% or more, or 75% or more, or 80% or more, or 85% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more sequence identity to all other members of the homology group.

[0038] In one embodiment, within each homology group, the gene sequence has 60% or more, or 65% or more, or 70% or more, or 75% or more, or 80% or more, or 85% or more to 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or less than 100% sequence identity to all other members of the homology group.

[0039] In one embodiment, the gene sequence of each homology group defines a protein sequence that is distinct in the homology group. In one embodiment, the distinct protein is selected from the group consisting of an S protein, an envelope protein, a nucleoprotein, and combinations thereof. In a further embodiment, the distinct protein is an S protein.

[0040] In a particular embodiment, multiple SARSrs are analyzed and classified into five homology groups, and within each homology group, gene sequences have greater than 65% to 99% sequence identity.

[0041] An exemplary process for identifying homology groups and consensus sequences is provided herein.

[0042] SARS-2β-CoV has a positive-sense single-stranded RNA genome of approximately 30 kb and four structural proteins. One of the structural proteins is the spike (S) peplomer. These S proteins are found on the surface of SARS-2β-CoV and mediate cellular receptor binding, thus determining the host tropism of the virus. The protein portion of the RNA encoding the S protein is divided into SI and S2 strands, with SI strand 10 and S2 strand 20 separated by a franc cleavage site 25, as shown in Figure 1. The RBD 30 is located in the SI strand 10. It was discovered that mutations in the RBD affect the binding of the virus to angiotensin-converting enzyme 2 (ACE2), and this enhanced binding is by facilitating the jump of the virus from animal hosts to human hosts. The membrane fusion portion 40 is located in the S2 strand 20. Further shown in Figure 1 are the heptapeptides HR1 and HR2, the transmembrane TM and the cytoplasmic domain of the S protein.

[0043] In contrast to other coronaviruses, such as SARS-1β-CoV, the S protein of SARS-2β-CoV is not enzymatically cleaved during virus assembly. The SARS-2β-CoV S protein is preactivated by the proprotein convertase furin, thus reducing the dependency on target cell proteases during cell entry.

[0044] The SARS-2β-CoV S protein is divided into S2 strand 10 and S2 strand 20. A conformational change in S2 strand 20 leads to viral fusion within the host cell. This, in combination with the S protein-encoding RNA sequence containing the RBD, makes the S protein-encoding RNA sequence an important candidate for use in anti-SARS-2β-CoV vaccine regimens.

[0045] FIG. 1 also shows the S protein portion of SARS-2 β-CoV that elicits a greater immune response (60). As shown, portion 60b overlaps with RBD 30 and is a significant portion, meaning that there is a significant immune response associated with RBD 30. Portions 60d and 60e overlap with the less variable membrane fusion portion 40, but are smaller and therefore elicit a less robust immune response. When analyzing immune responses against different β-CoVs, such as SARS-CoV-1 and MERS-CoV, it was observed that antibodies capable of neutralizing the activity of these coronaviruses could also bind to more conserved regions of the S protein, namely, within the S2 domain 20, within the S protein stem region.

[0046] As further illustrated in Figure 1, alignment of the S protein coding sequences of RNA from various SARSr shows significant differences throughout the gene (50). Therefore, a vaccine based on SARS-2β-CoV RNA of the present invention may not be able to efficiently protect against infection caused by other SARSr. However, analysis of the S protein coding sequences of RNA from multiple SARSr allows the SARSr to be classified into homology groups, as shown in Table 1. TIFF2025028831000002.tif63170

[0047] To obtain the information in Table 1, sequences were found using ViPR and NCBI. Global alignments were performed using Clustal Omega. Related alignments (>92%) were extracted to create groupings, which were aligned using Clustal Omega and confirmed using BLAST multiple sequence alignment.

[0048] For the SARS-CoV group, 1130 sequences from GenBank and ViPR covering the original SARS-CoV-1 were analyzed. Some sequences contained random inserts likely responsible for gaps, but a small number of variants all maintained antibody binding. For the SARS-CoV 2 group, over 3000 sequences were analyzed, including the new clades 20H, 201 and 20J (corresponding to the South African, California and UK variants, respectively). WIV-1 is the prominent SARSr in bats, but has been shown to replicate in human cells. 56 WIV-1 strains were analyzed, including the RaTG13 strain that is thought to have given rise to SARS-2β-CoV. Only 16 strains had a complete CDS. The structure appeared stable across variants, as indicated by the NCBI conserved protein domain families cd21477 and Cn3D. For the YNLF group, 71 sequences (39 with complete CDS) from bats, pangolins and camels. These SARSr strains are less similar to SARS-2β-CoV than to the WIV1 family, but have some strong similarities to the SARS-CoV and SARS-CoV 2 groups in certain regions. The overall spike alignment is moderate, but the RBD alignment shows strong similarities. In the Bat2013 group, 19 samples with high similarity were analyzed. The Bat2013 group shows higher variance than the other groups, but many strains show cross-reactivity to the same antibodies.

[0049] Consensus sequence According to an embodiment of the present disclosure, a method for preparing a vaccine for preventing at least one β-CoV infection in a subject, particularly a mammalian subject, more particularly a human subject, comprises identifying at least one consensus sequence for each homology group. A consensus sequence is a DNA, RNA or protein sequence developed for a group that contains the statistically most frequent residues at each position of the sequence. In one embodiment, the consensus sequence for a homology group has at least 65%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 99% commonality with each member of the corresponding homology group.

[0050] In certain embodiments, a consensus sequence is a DNA sequence that has 60% or more, or 65% or more, or 70% or more, or 75% or more, or 80% or more, or 85% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more sequence identity with all other members of the corresponding homology group.

[0051] In certain embodiments, a consensus sequence is an RNA sequence that has 60% or more, or 65% or more, or 70% or more, or 75% or more, or 80% or more, or 85% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more sequence identity with all other members of the corresponding homology group.

[0052] In certain embodiments, a consensus sequence is a protein sequence that has 60% or more, or 65% or more, or 70% or more, or 75% or more, or 80% or more, or 85% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more sequence identity with all other members of the corresponding homology group.

[0053] In one embodiment, the consensus sequence is edited to remove a variable domain. An exemplary variable domain is shown as sequence 324-533 in Figure 1. In embodiments where one or more variable domains are deleted, the deleted sequence is replaced by a smaller linker peptide designed to bridge the gap created by the deletion.

[0054] In one embodiment, the consensus sequence of each homology group is selected from the group consisting of a DNA sequence, an RNA sequence, a protein sequence, and combinations thereof. In one embodiment, the consensus sequence of at least one of the homology groups is RNA. In a further embodiment, the RNA is mRNA.

[0055] In one embodiment, the β-CoV analyzed is SARSr. In a further embodiment, the SARSr comprises at least one SARS-2β-CoV classified into at least one homology group, and the consensus sequence of at least one homology group is a DNA sequence, an RNA sequence, or a protein sequence. In an embodiment where a single SARSr, for example a single SARS-2β-CoV, is identified and where the single SARSr is the only member of the homology group, it will be understood that the consensus sequence may be a DNA sequence, and the RNA sequence or the protein sequence has 100% commonality with SARSr.

[0056] In one embodiment, the consensus sequence is a SARS-2β-CoV DNA sequence, which is at least a portion of the S protein coding sequence. In a further embodiment, the consensus sequence is a SARS-2β-CoV DNA sequence that includes the entire S protein coding sequence.

[0057] In one embodiment, the consensus sequence is a SARS-2β-CoV RNA sequence, which is at least a portion of the S protein coding sequence.

[0058] In one embodiment, the consensus sequence is a SARS-2β-CoV protein sequence, and the SARS-2β-CoV protein sequence is at least a portion of the S protein.

[0059] Viral Vectors According to an embodiment of the present disclosure, a method for preparing a vaccine for preventing at least one β-CoV infection in a subject, particularly a mammalian subject, more particularly a human subject, comprises inserting at least one consensus sequence into a viral vector. In one embodiment, the viral vector is an adenoviral vector element.

[0060] To minimize pre-existing and induced interfering anti-adenoviral immune responses, all endogenous genes are deleted from the adenoviral vector element, i.e., in one embodiment, the adenoviral vector element is a fully deleted (fd) adenoviral vector.

[0061] In one embodiment, the adenoviral vector 70, preferably the fd adenoviral vector, can accommodate gene constructs up to 33 kb, and has inverted terminal repeats (ITRs) 72, 72 and a packaging signal (Y) 73, as shown in Figure 2. The deleted endogenous genes are replaced with size-complementing stuffers 75. In the embodiment shown, these stuffers 75 are prepared from fragments of the human gene 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase gene (ATIC). In further embodiments, other stuffer sequences can be used, such as, but not limited to, human hypoxanthine-guanine phosphoribosyltransferase.

[0062] In the embodiment shown in Figure 2, five consensus sequences 80a, 80b, 80c, 80d, 80e are accepted by the viral vector or adenoviral vector or fd adenoviral vector. However, in further embodiments, the viral vector may contain more or fewer consensus sequences. The consensus sequences may be according to any embodiment or combination of embodiments provided herein.

[0063] For illustrative purposes only, and referring to FIG. 2, consensus sequence 80a is a SARS-2β-CoV RNA sequence derived from a homology group containing only SARS-2β-CoV. When the SARS-2β-CoV RNA sequence is the only consensus sequence included in the viral vector, the resulting vaccine is intended to provide specific protection from infection with SARS-2β-CoV. In further embodiments, the consensus sequence may be derived from a homology group that includes a broader collection of SARS-2β-CoVs. In still further embodiments, the viral vector may include additional consensus sequences derived from different homology groups, as shown in FIG. 2. In such embodiments, the resulting vaccine may provide broader protection against viruses of different homology groups.

[0064] In another embodiment, the viral vector can be a viral vector configured to deliver a transgene, such as a DNA transgene. Exemplary viral vectors configured to deliver a transgene include, but are not limited to, adenovirus-associated vectors and vaccinia virus vectors.

[0065] Packaging plasmids According to an embodiment of the present disclosure, a method for preparing a vaccine for preventing at least one β-CoV infection in a subject, particularly a mammalian subject, more particularly a human subject, comprises providing at least one packaging plasmid.

[0066] With further reference to Figure 2, an exemplary plasmid 82 used in the present disclosure includes multiple genes, such as late and early genes. In one embodiment, the plasmid 82 includes multiple late genes, preferably late regions 1, 2, 3, 4, and 6, as shown in Figure 2. In a further embodiment, the plasmid 82 used in the present disclosure includes multiple early genes, preferably early regions 2 and 4, as shown in Figure 2.

[0067] In one embodiment, the late and early genes are provided in trans.

[0068] Plasmid 82 further comprises the major late promoter (MLP) and the right ITR, as shown in Figure 2. However, the capsid used in this disclosure lacks the left ITR, the early genes El and E3, their packaging signals, and their protein IX gene.

[0069] In certain embodiments, the plasmid consists essentially of (i) late regions 1, 2, 3, 4, 5, (ii) early regions 2 and 4, (iii) the MLP, and (iv) the right ITR. In such embodiments, the plasmid completely lacks the left ITF, the early genes El and E3, the packaging signal, and the protein IX gene.

[0070] In one embodiment, plasmid 82 is based on an adenovirus. In a further embodiment, plasmid 82 is based on an adenovirus selected from the group consisting of Ad2, Ad5, Ad6 and Ad35 serotypes and combinations thereof, with the adenovirus capsid of human serotype Ad2 being encoded by pPaC2, Ad5 with pPaC5, Ad6 with pPaC6 and Ad35 with pPaB35. Transfection

[0071] According to an embodiment of the present disclosure, a method for preparing a vaccine for preventing at least one β-CoV infection in a subject, particularly a mammalian subject, more particularly a human subject, comprises transfecting a packaging cell with a viral vector and a packaging plasmid. In one embodiment, the packaging cell may contain one or more viral vectors and one or more plasmids. In a preferred embodiment, the packaging cell contains at least one, preferably two or more, more preferably three or more viral vectors and one packaging plasmid.

[0072] 2, viral vector 70 and plasmid 82 are introduced into eukaryotic host cells or packaging cells 85 by co-transfection. In the particular embodiment shown, viral vector 70 and plasmid 82 are co-transfected into packaging cells 85 using an optimized and standardized one-week co-transfection protocol.

[0073] In one embodiment, the viral vector is an adenoviral vector, particularly an fd adenoviral vector, and the packaging cells are derived from cell lines such as, but not limited to, human embryonic kidney cells (HEK293) and PerC.6 cells. The packaging cells required to package the fd adenoviral vector must be engineered to express the genes encoded within the El region of the adenoviral vector. In a particular embodiment, the packaging cells are HEK293-derived Q7 packaging cells engineered to express the genes encoded within the E1 region of the adenoviral vector.

[0074] It is noteworthy that the production of fd adenoviral vectors is initiated by chemical transfection of packaging cells with a mixture of an engineered adenoviral genome, a packaging expression plasmid and a chemical transfection reagent.

[0075] Capsid According to an embodiment of the present disclosure, a method for preparing a vaccine for preventing at least one β-CoV infection in a subject, particularly a mammalian subject, more particularly a human subject, includes encapsidating a packaging cell 85 containing a viral vector 70 and a plasmid 82 into a capsid, as shown in FIG.

[0076] The packaging cell 85 containing the viral vector 70 and the plasmid 82 is delivered in a capsid of an adenovirus serotype that is rare in the mammal being vaccinated. In a particular embodiment, the mammal being vaccinated is a human and the viral vector is delivered in a capsid of an adenovirus serotype that is rare in humans. In one embodiment, the viral vector is delivered in a capsid of the Ad2, Ad5, Ad6 and Ad35 serotypes, and combinations thereof. In one embodiment, the viral vector is delivered in a capsid of the Ad6 serotype.

[0077] Vaccine composition In one embodiment, the present disclosure provides a composition of a vaccine, more specifically a vaccine for preventing infection from a β-CoV, preferably SARSr.

[0078] According to an embodiment of the present disclosure, the vaccine comprises one or more consensus sequences derived from one or more β-CoVs, preferably one or more SARSr, carried by at least one viral vector. The consensus sequences can be according to any embodiment or combination of embodiments described herein. The viral vector can be according to any embodiment or combination of embodiments described above.

[0079] In one embodiment, the one or more consensus sequences are β-CoV DNA sequences, RNA sequences, protein sequences, or a combination thereof, preferably SARSr DNA sequences, RNA sequences, protein sequences, or a combination thereof.

[0080] According to embodiments of the present disclosure, the one or more consensus sequences include at least one SARSr DNA or RNA sequence, or preferably at least one SARS-2β-CoV DNA or RNA sequence, hi one embodiment, the SARSr DNA or RNA sequence, or the SARS-2β-CoV DNA or RNA sequence, is at least a portion of the S protein coding sequence.

[0081] In some embodiments, one or more of the one or more consensus sequences have a variable region partially or completely removed. In certain embodiments, the one or more consensus sequences comprise at least one SARSr DNA or RNA sequence, preferably at least one SARS-2β-CoV DNA or RNA sequence, which is at least a portion of the S protein coding sequence, and at least a portion of the variable region of the S protein coding sequence is removed.

[0082] In one embodiment, expression of the consensus sequence is driven by a promoter. The promoter can be specific to the consensus sequence, the vaccinated animal, and the particular composition of the vaccine. In one embodiment, the promoter is selected from the group consisting of human cytomegalovirus immediate early promoter / enhancer, polyadenylation site from human growth gene, elongation factor 1-alpha, phosphoglycerate kinase, ubiquitin C, beta actin gene, and combinations thereof. In an embodiment, the activity of the promoter can be affected by chemicals, such as, but not limited to, antibiotics. Tetracycline is a non-limiting example of an antibiotic that affects the activity of the promoter.

[0083] In certain embodiments, the vaccine is specifically designed to prevent infection from at least SARS-2β-CoV. In such embodiments, the one or more consensus sequences comprise at least one SARS-2β-CoV DNA or RNA sequence. Preferably, the SARS-2β-CoV DNA or RNA sequence is a DNA or RNA sequence that encodes the S protein. In further embodiments, the SARS-2β-CoV DNA or RNA sequence is an RNA sequence that is (partially or entirely) a sequence that encodes the S protein.

[0084] In embodiments where the consensus sequence is a SARS-2β-CoV RNA sequence encoding (partially or entirely) the S protein, the SARS-2β-CoV DNA sequence is human codon optimized and expression of the specific RNA is driven by the human cytomegalovirus immediate early promoter / enhancer followed by a polyadenylation site derived from the human growth factor gene. In other embodiments, expression of the SARS-2β-CoV RNA is driven by other promoters, including but not limited to promoters derived from elongation factor 1-alpha, phosphoglycerate kinase, ubiquitin C, β-actin genes, and combinations thereof. In another embodiment, expression of the SARS-2β-CoV RNA is driven by a promoter whose activity can be affected by chemicals, including but not limited to the antibiotic tetracycline.

[0085] In further embodiments, the vaccine comprises two or more consensus sequences, one or more viral vectors. According to embodiments of the present disclosure, one consensus sequence is a SARS-2β-CoV DNA or RNA sequence, and the vaccine comprises at least one additional consensus sequence that is a SARSr DNA, RNA or protein sequence.

[0086] In one embodiment, the at least one viral vector is an adenoviral vector, more preferably an fd adenoviral vector.

[0087] In a further embodiment, the vaccine is a SARSr vaccine comprising a viral vector having all or part of a SARS-2β-CoV RNA sequence and at least one other SARSr RNA (all or part) sequence. In such an embodiment, the viral vector also carries an expression cassette for the human codon-optimized S protein for each of the SARSr members represented in the viral vector. The human codon-optimized S protein is driven by the CMV immediate early promoter / enhancer followed by a polyadenylation site derived from human growth hormone.

[0088] In some embodiments, the SARS-2β-CoV RNA and, if present, the additional SARSr RNA have the variable region of the S protein coding sequence completely or partially deleted.

[0089] According to embodiments of the present disclosure, the vaccine further comprises a packaging plasmid. The packaging plasmid can be according to any embodiment or combination of embodiments described herein.

[0090] In one embodiment, the at least one consensus sequence is a SARSr DNA or RNA sequence, in particular a SARSr DNA or RNA sequence that is an S protein coding sequence, and the packaging plasmid lacks the left ITR, the early genes E1 and E3, its packing signal, and its protein IX gene. In a particular embodiment, the at least one consensus sequence is a SARSr DNA or RNA sequence, in particular a SARSr DNA or RNA sequence that is an S protein coding sequence, contained in a viral vector and a packaging plasmid based on an adenovirus selected from the group consisting of serotypes Ad2, Ad5, Ad6 and Ad35 and combinations thereof, the adenovirus capsid of human serotype Ad2 is encoded by pPaC2, Ad5 with pPaC5, Ad6 with pPaC6 and Ad35 with pPaB35, and the plasmid lacks the left ITR, the early genes E1 and E3, its packing signal, and its protein IX gene.

[0091] According to embodiments of the present disclosure, the vaccine comprises packaging cells co-transfected with a consensus-containing viral vector and a plasmid, the packaging being according to any embodiment or combination or embodiments disclosed herein.

[0092] In one embodiment, the viral vector and the plasmid are co-transfected into the packaging cells using an optimized and standardized one-week co-transfection protocol using HEK-293 derived Q7 packaging cells. In an exemplary embodiment, the viral vector comprises at least one consensus sequence comprising a SARSr DNA or RNA sequence, in particular a sequence encoding the S protein, and the plasmid is based on an adenovirus selected from the group consisting of serotypes Ad2, Ad5, Ad6 and Ad35 and combinations thereof, and the adenovirus capsid of human serotype Ad2 is encoded by pPaC2, Ad5 with pPaC5, Ad6 with pPaC6 and Ad35 with pPaB35, and the viral vector and the plasmid are co-transfected into the packaging cells using an optimized and standardized one-week co-transfection protocol using HEK-293 derived Q7 packaging cells.

[0093] The vaccine comprises a capsid in which the packaging cell is encapsidated (along with the viral vector and plasmid). The capsid can be according to any embodiment or combination of embodiments disclosed herein.

[0094] In one embodiment, the capsid is of the Ad2, Ad5, Ad6 and Ad35 serotypes, and combinations thereof.

[0095] Vaccination Method In one embodiment, the present disclosure provides a method of vaccinating an animal subject, preferably a mammalian subject, more preferably a human subject, against infection from at least one group of β-CoV.

[0096] According to an embodiment of the present disclosure, the method includes providing a vaccine comprising at least one viral vector and a plasmid comprising at least one β-CoV consensus sequence, preferably at least one SARSr consensus sequence, more preferably at least one SARS-2 β-CoV consensus sequence, wherein the at least one viral vector and the plasmid are transfected into a packaging cell, and the packaging cell is encapsidated into a capsid.

[0097] In one embodiment, at least one β-CoV consensus sequence is according to any embodiment or combination or embodiments described herein. In one embodiment, at least one viral vector is according to any embodiment or combination of embodiments described herein. In one embodiment, the plasmid is according to any embodiment or combination of embodiments described herein. In one embodiment, the packaging cell is according to any embodiment or combination of embodiments described herein. In one embodiment, the capsid is according to any embodiment or combination of embodiments described herein.

[0098] The method further comprises injecting the viral vector into an animal subject, preferably a mammalian subject, such as a human. In one embodiment, the single dose is sufficient to provide protection against at least one β-CoV, more specifically, any β-CoV that has 60% or more, or 65% or more, or 70% or more, or 75% or more, or 80% or more, or 85% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more sequence identity to at least one of the consensus sequences included in the vaccine.

[0099] In other embodiments, more than one dose may be required to provide protection. In particular, two, three, or four doses are sufficient to provide protection against at least one β-CoV, more particularly any β-CoV that has 60% or more, or 65% or more, or 70% or more, or 75% or more, or 80% or more, or 85% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more sequence identity to at least one of the consensus sequences included in the vaccine. EXAMPLES

[0100] Example 1 To demonstrate the efficiency of the viral vector according to an embodiment of the present disclosure, BALB / c mice were administered various doses of A / Vietnam / 1203 / 2004 (H5N1) vaccine using a viral vector according to an embodiment of the present disclosure and then challenged with H5N1 virus. In particular, there were four groups of 10 mice each. The first control group (Cl) is vaccinated with placebo. The second control group (C2) is not vaccinated. The first experimental group (El) is vaccinated with 3×10 IgG1-specific ... 8 The second experimental group (E2) will be vaccinated with a genome-equivalent amount of GreFluVie vaccine (containing a viral vector containing a consensus sequence with at least 60% commonality with the H5N1 virus). 7 Mice were vaccinated with a genome equivalent amount of GreFluVie vaccine. Groups Cl, E1 and E2 were boosted on day 24 with the same control or vaccine preparation. On day 26, groups Cl, E1 and E2 were challenged intranasally with a median lethal dose (LD50) of H5N1. The groups were observed and their weights were measured daily. On day 48, mice were bled and tested for the presence of antibodies that neutralize infection of MDCK test cells (MDCK test sells) by the H5N1 virus and that inhibit hemagglutination of horse red blood cells.

[0101] As shown in Figure 3, the Cl group has a very poor survival rate, with all mice dying before 15 days after infection. In contrast, both E1 and E2 groups show significantly improved survival rates, with body weights mimicking the trend of the C2 group. Among the E1 and E2 groups, the E1 group shows higher virus neutralization and lower lung virus titers. Importantly, both E1 and E2 groups show a significant improvement in the ability to fight infection after immunization.

[0102] Example 2 A control group (C3) of 5 mice (BALB / c mice) will be vaccinated with a placebo. An experimental group (E3) of 5 mice (BALB / c mice) will be vaccinated with 3 × 10 7 The mice were vaccinated with a genome equivalent of the GreMERSfl vaccine (containing a viral vector with a consensus sequence with at least 60% commonality with EMX / 2012 MERS-CoV). The consensus sequence is specifically the full-length spike protein of MERS-CoV. Groups C3 and E3 were boosted on day 17 with the same control or vaccine preparation. On day 19, groups C3 and E3 were intranasally infected with LD50 of MERS. The groups were mixed on day 21. Serum was tested for the presence of antibodies that neutralize infection of test cells with EMX / 2012 MERS-CoV.

[0103] As shown in Figure 4, the E3 group showed a significant improvement in virus neutralization.

[0104] Although several embodiments of the viral vectors and associated vaccines have been described in detail herein, it is clear that they can be engineered and modified, all of which are within the true spirit and scope of the present invention. In particular, the present viral vectors and vaccines have been described in detail with respect to β-CoV, and more specifically SARS-2 and SARSr viruses, but it will be understood that the viral vectors and vaccines can be engineered according to the techniques of those skilled in the art to apply to other classes of coronaviruses, such as, for example, a-CoV, g-CoV, and d-CoV. Moreover, since numerous modifications and changes will readily occur to those skilled in the art, it is not desired to limit the invention to the exact construction and operation shown and described, and therefore, all suitable modifications and equivalents may be used and are within the scope of the present disclosure. (Sequence Listing) SEQUENCE LISTING <110> Greffex <120> Engineering Broadly Reactive Coronavirus Vaccines, Their Design and Uses <130> 543346.14 <150> US 63 / 012360 <151> 2020-04-20 <160> 5 <170> PatentIn version 3.5 <210> 1 <211> 1255 <212> PRT <213> SARS-CoV <400> 1 Met Phe Ile Phe Leu Leu Phe Leu Thr Leu Thr Ser Gly Ser Asp Leu 1 5 10 15 Asp Arg Cys Thr Thr Phe Asp Asp Val Gln Ala Pro Asn Tyr Thr Gln 20 25 30 His Thr Ser Ser Met Arg Gly Val Tyr Tyr Pro Asp Glu Ile Phe Arg 35 40 45 Ser Asp Thr Leu Tyr Leu Thr Gln Asp Leu Phe Leu Pro Phe Tyr Ser 50 55 60 Asn Val Thr Gly Phe His Thr Ile Asn His Thr Phe Gly Asn Pro Val 65 70 75 80 Ile Pro Phe Lys Asp Gly Ile Tyr Phe Ala Ala Thr Glu Lys Ser Asn 85 90 95 Val Val Arg Gly Trp Val Phe Gly Ser Thr Met Asn Asn Lys Ser Gln 100 105 110 Ser Val Ile Ile Ile Asn Asn Ser Thr Asn Val Val Ile Arg Ala Cys 115 120 125 Asn Phe Glu Leu Cys Asp Asn Pro Phe Phe Ala Val Ser Lys Pro Met 130 135 140 Gly Thr Gln Thr His Thr Met Ile Phe Asp Asn Ala Phe Asn Cys Thr 145 150 155 160 Phe Glu Tyr Ile Ser Asp Ala Phe Ser Leu Asp Val Ser Glu Lys Ser 165 170 175 Gly Asn Phe Lys His Leu Arg Glu Phe Val Phe Lys Asn Lys Asp Gly 180 185 190 Phe Leu Tyr Val Tyr Lys Gly Tyr Gln Pro Ile Asp Val Val Arg Asp 195 200 205 Leu Pro Ser Gly Phe Asn Thr Leu Lys Pro Ile Phe Lys Leu Pro Leu 210 215 220 Gly Ile Asn Ile Thr Asn Phe Arg Ala Ile Leu Thr Ala Phe Ser Pro 225 230 235 240 Ala Gln Asp Ile Trp Gly Thr Ser Ala Ala Ala Tyr Phe Val Gly Tyr 245 250 255 Leu Lys Pro Thr Thr Phe Met Leu Lys Tyr Asp Glu Asn Gly Thr Ile 260 265 270 Thr Asp Ala Val Asp Cys Is Gln Asn Pro Leu Ala Glu Leu Lys Cys 275 280 285 Ser Val Lys Ser Phe Glu Ile Asp Lys Gly Ile Tyr Gln Thr Ser Asn 290,295,300 Phe Arg Val Val Pro Ser Gly Asp Val Val Arg Phe Pro Asn Ile Thr 305 310 315 320 Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Pro Ser 325 330 335 Val Tyr Only Trp Glu Arg Lys Lys Ile Ser Asn Cys Only Only Asp Tyr 340 345 350 Ser Val Leu Tyr Asn Ser Thr Phe Phe Ser Thr Phe Lys Cys Tyr Gly 355 360 365 Val Ser Ala Thr Lys Leu Asn Asp Leu Cys Phe Ser Asn Val Tyr Ala 370 375 380 Asp Ser Phe Val Val Lys Gly Asp Asp Val Arg Gln Ile Ala Pro Gly 385 390 395 400 Gln Thr Gly Val Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe 405 410 415 Met Gly Cys Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr Ser 420 425 430 Thr Gly Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg His Gly Lys Leu 435 440 445 Arg Pro Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp Gly 450 455 460 Lys Pro Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp Pro Leu Asn Asp 465 470 475 480 Tyr Gly Phe Tyr Thr Thr Thr Gly Ile Gly Tyr Gln Pro Tyr Arg Val 485 490 495 Val Val Leu Ser Phe Glu Leu Leu Asn Ala Pro Ala Thr Val Cys Gly 500 505 510 Pro Lys Leu Ser Thr Asp Leu Ile Lys Asn Gln Cys Val Asn Phe Asn 515 520 525 Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Pro Ser Ser Lys Arg 530 535 540 Phe Gln Pro Phe Gln Gln Phe Gly Arg Asp Val Ser Asp Phe Thr Asp 545 550 555 560 Ser Val Arg Asp Pro Lys Thr Ser Glu Ile Leu Asp Ile Ser Pro Cys 565 570 575 Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Ala Ser Ser 580 585 590 Glu Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Asp Val Ser Thr 595 600 605 Ala Ile His Ala Asp Gln Leu Thr Pro Ala Trp Arg Ile Tyr Ser Thr 610 615 620 Gly Asn Asn Val Phe Gln Thr Gln Ala Gly Cys Leu Ile Gly Ala Glu 625 630 635 640 His Val Asp Thr Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile 645 650 655 Cys Ala Ser Tyr His Thr Val Ser Leu Leu Arg Ser Thr Ser Gln Lys 660 665 670 Ser Ile Val Ala Tyr Thr Met Ser Leu Gly Ala Asp Ser Ser Ile Ala 675 680 685 Tyr Ser Asn Asn Thr Ile Ala Ile Pro Thr Asn Phe Ser Ile Ser Ile 690 695 700 Thr Thr Glu Val Met Pro Val Ser Met Ala Lys Thr Ser Val Asp Cys 705 710 715 720 Asn Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ala Asn Leu Leu Leu 725 730 735 Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Ser Gly Ile 740 745 750 Ala Ala Glu Gln Asp Arg Asn Thr Arg Glu Val Phe Ala Gln Val Lys 755 760 765 Gln Met Tyr Lys Thr Pro Thr Leu Lys Tyr Phe Gly Gly Phe Asn Phe 770 775 780 Ser Gln Ile Leu Pro Asp Pro Leu Lys Pro Thr Lys Arg Ser Phe Ile 785 790 795 800 Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Met 805 810 815 Lys Gln Tyr Gly Glu Cys Leu Gly Asp Ile Asn Ala Arg Asp Leu Ile 820 825 830 Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr 835 840 845 Asp Asp Met Ile Ala Ala Tyr Thr Ala Ala Leu Val Ser Gly Thr Ala 850 855 860 Thr Ala Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe 865 870 875 880 Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn 885 890 895 Val Leu Tyr Glu Asn Gln Lys Gln Ile Ala Asn Gln Phe Asn Lys Ala 900 905 910 Ile Ser Gln Ile Gln Glu Ser Leu Thr Thr Thr Ser Thr Ala Leu Gly 915 920 925 Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu 930 935 940 Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn 945 950 955 960 Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln Ile Asp 965 970 975 Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln 980 985 990 Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala 995 1000 1005 Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp 1010 1015 1020 Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ala Ala 1025 1030 1035 Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ser Gln 1040 1045 1050 Glu Arg Asn Phe Thr Thr Ala Pro Ala Ile Cys His Glu Gly Lys 1055 1060 1065 Ala Tyr Phe Pro Arg Glu Gly Val Phe Val Phe Asn Gly Thr Ser 1070 1075 1080 Trp Phe Ile Thr Gln Arg Asn Phe Phe Ser Pro Gln Ile Ile Thr 1085 1090 1095 Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly 1100 1105 1110 Ile Ile Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp 1115 1120 1125 Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser 1130 1135 1140 Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val 1145 1150 1155 Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys 1160 1165 1170 Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr 1175 1180 1185 Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Val Trp Leu Gly Phe Ile 1190 1195 1200 Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Leu Leu Cys Cys 1205 1210 1215 Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Ala Cys Ser Cys Gly 1220 1225 1230 Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys 1235 1240 1245 Gly Val Lys Leu His Tyr Thr 1250 1255 <210> 2 <211> 1256 <212> PRT <213> SARSr <400> 2 Met Lys Leu Leu Val Leu Val Phe Ala Thr Leu Val Ser Ser Tyr Thr 1 5 10 15 Ile Glu Lys Cys Leu Asp Phe Asp Asp Arg Thr Pro Pro Ala Asn Thr 20 25 30 Gln Phe Leu Ser Ser His Arg Gly Val Tyr Tyr Pro Asp Asp Ile Phe 35 40 45 Arg Ser Asn Val Leu His Leu Val Gln Asp His Phe Leu Pro Phe Asp 50 55 60 Ser Asn Val Thr Arg Phe Ile Thr Phe Gly Leu Asn Phe Asp Asn Pro 65 70 75 80 Ile Ile Pro Phe Lys Asp Gly Ile Tyr Phe Ala Ala Thr Glu Lys Ser 85 90 95 Asn Val Ile Arg Gly Trp Val Phe Gly Ser Thr Met Asn Asn Lys Ser 100 105 110 Gln Ser Val Ile Ile Met Asn Asn Ser Thr Asn Leu Val Ile Arg Ala 115 120 125 Cys Asn Phe Glu Leu Cys Asp Asn Pro Phe Phe Val Val Leu Lys Ser 130 135 140 Asn Asn Thr Gln Ile Pro Ser Tyr Ile Phe Asn Asn Ala Phe Asn Cys 145 150 155 160 Thr Phe Glu Tyr Val Ser Lys Asp Phe Asn Leu Asp Leu Gly Glu Lys 165 170 175 Pro Gly Asn Phe Lys Asp Leu Arg Glu Phe Val Phe Arg Asn Lys Asp 180 185 190 Gly Phe Leu His Val Tyr Ser Gly Tyr Gln Pro Ile Ser Ala Ala Ser 195 200 205 Gly Leu Pro Thr Gly Phe Asn Ala Leu Lys Pro Ile Phe Lys Leu Pro 210 215 220 Leu Gly Ile Asn Ile Thr Asn Phe Arg Thr Leu Leu Thr Ala Phe Pro 225 230 235 240 Pro Arg Pro Asp Tyr Trp Gly Thr Ser Ala Ala Ala Tyr Phe Val Gly 245 250 255 Tyr Leu Lys Pro Thr Thr Phe Met Leu Lys Tyr Asp Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ser Gln Asn Pro Leu Ala Glu Leu Lys 275 280 285 Cys Ser Val Lys Ser Phe Glu Ile Asp Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Ala Pro Ser Lys Glu Val Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Thr Phe Pro 325 330 335 Ser Val Tyr Ala Trp Glu Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Thr Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Ala Thr Lys Leu Asn Asp Leu Cys Phe Ser Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Val Lys Gly Asp Asp Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Val Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr 420 425 430 Gln Thr Gly Asn Tyr Asn Tyr Lys Tyr Arg Ser Leu Arg His Gly Lys 435 440 445 Leu Arg Pro Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp 450 455 460 Gly Lys Pro Cys Thr Pro Pro Ala Phe Asn Cys Tyr Trp Pro Leu Asn 465 470 475 480 Asp Tyr Gly Phe Tyr Ile Thr Asn Gly Ile Gly Tyr Gln Pro Tyr Arg 485 490 495 Val Val Val Leu Ser Phe Glu Leu Leu Asn Ala Pro Ala Thr Val Cys 500 505 510 Gly Pro Lys Leu Ser Thr Asp Leu Ile Lys Asn Gln Cys Val Asn Phe 515 520 525 Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Pro Ser Ser Lys 530 535 540 Arg Phe Gln Pro Phe Gln Gln Phe Gly Arg Asp Val Ser Asp Phe Thr 545 550 555 560 Asp Ser Val Arg Asp Pro Lys Thr Ser Glu Ile Leu Asp Ile Ser Pro 565 570 575 Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser 580 585 590 Ser Glu Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Asp Val Pro 595 600 605 Val Ala Ile His Ala Asp Gln Leu Thr Pro Ser Trp Arg Val Tyr Ser 610 615 620 Thr Gly Asn Asn Val Phe Gln Thr Gln Ala Gly Cys Leu Ile Gly Ala 625 630 635 640 Glu His Val Asp Thr Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly 645 650 655 Ile Cys Ala Ser Tyr His Thr Val Ser Ser Leu Arg Ser Thr Ser Gln 660 665 670 Lys Ser Ile Val Ala Tyr Thr Met Ser Leu Gly Ala Asp Ser Ser Ile 675 680 685 Ala Tyr Ser Asn Asn Thr Ile Ala Ile Pro Thr Asn Phe Ser Ile Ser 690 695 700 Ile Thr Thr Glu Val Met Pro Val Ser Met Ala Lys Thr Ser Val Asp 705 710 715 720 Cys Asn Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ala Asn Leu Leu 725 730 735 Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Ser Gly 740 745 750 Ile Ala Val Glu Gln Asp Arg Asn Thr Arg Glu Val Phe Ala Gln Val 755 760 765 Lys Gln Met Tyr Lys Thr Pro Thr Leu Lys Asp Phe Gly Gly Phe Asn 770 775 780 Phe Ser Gln Ile Leu Pro Asp Pro Leu Lys Pro Thr Lys Arg Ser Phe 785 790 795 800 Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe 805 810 815 Met Lys Gln Tyr Gly Glu Cys Leu Gly Asp Ile Asn Ala Arg Asp Leu 820 825 830 Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu 835 840 845 Thr Asp Asp Met Ile Ala Ala Tyr Thr Ala Ala Leu Val Ser Gly Thr 850 855 860 Ala Thr Ala Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro 865 870 875 880 Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln 885 890 895 Asn Val Leu Tyr Glu Asn Gln Lys Gln Ile Ala Asn Gln Phe Asn Lys 900 905 910 Ala Ile Ser Gln Ile Gln Glu Ser Leu Thr Thr Thr Ser Thr Ala Leu 915 920 925 Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr 930 935 940 Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu 945 950 955 960 Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln Ile 965 970 975 Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr 980 985 990 Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala 995 1000 1005 Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val 1010 1015 1020 Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ala 1025 1030 1035 Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ser 1040 1045 1050 Gln Glu Arg Asn Phe Thr Thr Ala Pro Ala Ile Cys His Glu Gly 1055 1060 1065 Lys Ala Tyr Phe Pro Arg Glu Gly Val Phe Val Phe Asn Gly Thr 1070 1075 1080 Ser Trp Phe Ile Thr Gln Arg Asn Phe Phe Ser Pro Gln Ile Ile 1085 1090 1095 Thr Thr Asp Asn Thr Phe Val Ser Gly Ser Cys Asp Val Val Ile 1100 1105 1110 Gly Ile Ile Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu 1115 1120 1125 Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr 1130 1135 1140 Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser 1145 1150 1155 Val Val Asn Ile Gln Lys Glu Ile Asn Arg Leu Asn Glu Val Ala 1160 1165 1170 Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys 1175 1180 1185 Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Val Trp Leu Gly Phe 1190 1195 1200 Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Leu Leu Cys 1205 1210 1215 Cys Met Thr Ser Cys Ser Cys Leu Lys Gly Ala Cys Ser Cys 1220 1225 1230 Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu 1235 1240 1245 Lys Gly Val Lys Leu His Tyr Thr 1250 1255 <210> 3 <211> 1241 <212> PRT <213> SARS <400> 3 Met Lys Ile Leu Ile Phe Ala Phe Leu Val Thr Leu Val Lys Ala Gln 1 5 10 15 Glu Gly Cys Gly Leu Ile Ser Arg Lys Pro Gln Pro Lys Met Ala Gln 20 25 30 Val Ser Ser Ser Arg Arg Gly Val Tyr Tyr Asn Asp Asp Ile Phe Arg 35 40 45 Ser Asp Val Leu His Leu Thr Gln Asp Tyr Phe Leu Pro Phe Asp Ser 50 55 60 Asn Leu Thr Gln Tyr Phe Ser Leu Asn Val Asp Ser Asp Arg Tyr Thr 65 70 75 80 Tyr Phe Asp Asn Pro Ile Leu Asp Phe Gly Asp Gly Val Tyr Phe Ala 85 90 95 Ala Thr Glu Lys Ser Asn Val Ile Arg Gly Trp Ile Phe Gly Ser Thr 100 105 110 Phe Asp Asn Thr Thr Gln Ser Ala Val Ile Val Asn Asn Ser Thr His 115 120 125 Ile Ile Ile Arg Val Cys Asn Phe Asn Leu Cys Lys Glu Pro Met Tyr 130 135 140 Thr Val Ser Arg Gly Thr Gln Gln Ser Ser Trp Val Tyr Gln Ser Ala 145 150 155 160 Phe Asn Cys Thr Tyr Asp Arg Val Glu Arg Ser Phe Gln Leu Asp Thr 165 170 175 Ala Pro Lys Thr Gly Asn Phe Lys Asp Leu Arg Glu Tyr Val Phe Lys 180 185 190 Asn Arg Asp Gly Phe Leu Ser Val Tyr Gln Thr Tyr Thr Ala Val Asn 195 200 205 Leu Pro Arg Gly Leu Pro Thr Gly Phe Ser Val Leu Arg Pro Ile Leu 210 215 220 Lys Leu Pro Phe Gly Ile Asn Ile Thr Ser Tyr Arg Val Val Met Ala 225 230 235 240 Met Phe Ser Gln Thr Thr Ser Asn Phe Leu Pro Glu Ser Ala Ala Tyr 245 250 255 Tyr Val Gly Asn Leu Lys Tyr Thr Thr Phe Met Leu Ser Phe Asn Glu 260 265 270 Asn Gly Thr Ile Thr Asp Ala Ile Asp Cys Ala Gln Asn Pro Leu Ala 275 280 285 Glu Leu Lys Cys Thr Ile Lys Asn Phe Asn Val Ser Lys Gly Ile Tyr 290 295 300 Gln Thr Ser Asn Phe Arg Val Ser Pro Thr Gln Glu Val Ile Arg Phe 305 310 315 320 Pro Asn Ile Thr Asn Arg Cys Pro Phe Asp Lys Val Phe Asn Ala Ser 325 330 335 Arg Phe Pro Asn Val Tyr Ala Trp Glu Arg Thr Lys Ile Ser Asp Cys 340 345 350 Val Ala Asp Tyr Thr Val Leu Tyr Asn Ser Thr Ser Phe Ser Thr Phe 355 360 365 Lys Cys Tyr Gly Val Ser Pro Ser Lys Leu Ile Asp Leu Cys Phe Thr 370 375 380 Ser Val Tyr Ala Asp Thr Phe Leu Ile Arg Ser Ser Glu Val Arg Gln 385 390 395 400 Val Ala Pro Gly Glu Thr Gly Val Ile Ala Asp Tyr Asn Tyr Lys Leu 405 410 415 Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Thr Ala Lys Gln 420 425 430 Asp Gln Gly Gln Tyr Tyr Tyr Arg Ser Ser Arg Lys Thr Lys Leu Lys 435 440 445 Pro Phe Glu Arg Asp Leu Ser Ser Asp Glu Asn Gly Val Arg Thr Leu 450 455 460 Ser Thr Tyr Asp Phe Tyr Pro Thr Val Pro Ile Glu Tyr Gln Ala Thr 465 470 475 480 Arg Val Val Val Leu Ser Phe Glu Leu Leu Asn Ala Pro Ala Thr Val 485 490 495 Cys Gly Pro Lys Leu Ser Thr Gly Leu Val Lys Asn Gln Cys Val Asn 500 505 510 Phe Asn Phe Asn Gly Leu Lys Gly Thr Gly Val Leu Thr Asp Ser Ser 515 520 525 Lys Arg Phe Gln Ser Phe Gln Gln Phe Gly Arg Asp Met Ser Asp Phe 530 535 540 Thr Asp Ser Val Arg Asp Pro Gln Thr Leu Gln Ile Leu Asp Ile Thr 545 550 555 560 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Ala 565 570 575 Ser Ser Glu Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Asp Val 580 585 590 Pro Thr Ala Ile Arg Ala Asp Gln Leu Thr Pro Ala Trp Arg Val Tyr 595 600 605 Ser Thr Gly Ile Asn Val Phe Gln Thr Gln Ala Gly Cys Leu Ile Gly 610 615 620 Ala Glu His Val Asn Ala Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 625 630 635 640 Gly Ile Cys Ala Ser Tyr His Thr Ala Ser Thr Leu Arg Ser Val Gly 645 650 655 Gln Lys Ser Ile Val Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 660 665 670 Ile Ala Tyr Ala Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Ser Ile 675 680 685 Ser Val Thr Thr Glu Val Met Pro Val Ser Met Ser Lys Thr Ser Val 690 695 700 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Gln Glu Cys Ser Asn Leu 705 710 715 720 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 725 730 735 Gly Ile Ala Ile Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 740 745 750 Val Lys Gln Met Tyr Lys Thr Pro Ala Ile Lys Asp Phe Gly Gly Phe 755 760 765 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Thr Lys Arg Ser 770 775 780 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 785 790 795 800 Phe Met Lys Gln Tyr Gly Glu Cys Leu Gly Asp Ile Asn Ala Arg Asp 805 810 815 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 820 825 830 Leu Thr Asp Asp Met Ile Ala Ala Tyr Thr Ala Ala Leu Val Ser Gly 835 840 845 Thr Ala Thr Ala Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 850 855 860 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 865 870 875 880 Gln Asn Val Leu Tyr Glu Asn Gln Lys Gln Ile Ala Asn Gln Phe Asn 885 890 895 Lys Ala Ile Ser Gln Ile Gln Glu Ser Leu Thr Thr Thr Ser Thr Ala 900 905 910 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 915 920 925 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 930 935 940 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 945 950 955 960 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 965 970 975 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 980 985 990 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val 995 1000 1005 Asp Phe Cys Gly Arg Gly Tyr His Leu Met Ser Phe Pro Gln Ala 1010 1015 1020 Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ser 1025 1030 1035 Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Glu Gly 1040 1045 1050 Lys Ala Tyr Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr 1055 1060 1065 Ser Trp Phe Ile Thr Gln Arg Asn Phe Tyr Ser Pro Gln Ile Ile 1070 1075 1080 Thr Thr Asp Asn Thr Phe Val Ala Gly Asn Cys Asp Val Val Ile 1085 1090 1095 Gly Ile Ile Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu 1100 1105 1110 Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr 1115 1120 1125 Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser 1130 1135 1140 Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala 1145 1150 1155 Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys 1160 1165 1170 Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Val Trp Leu Gly Phe 1175 1180 1185 Ile Ala Gly Leu Ile Ala Ile Val Met Ala Thr Ile Leu Leu Cys 1190 1195 1200 Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Ala Cys Ser Cys 1205 1210 1215 Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu 1220 1225 1230 Lys Gly Val Lys Leu His Tyr Thr 1235 1240 <210> 4 <211> 1246 <212> PRT <213> SARS <400> 4 Met Leu Phe Phe Leu Phe Leu Gln Phe Ala Leu Val Asn Ser Gln Cys 1 5 10 15 Val Asn Leu Thr Gly Arg Thr Pro Leu Asn Pro Asn Tyr Thr Asn Ser 20 25 30 Ser Gln Arg Gly Val Tyr Tyr Pro Asp Thr Ile Tyr Arg Ser Asp Thr 35 40 45 Leu Val Leu Ser Gln Gly Tyr Phe Leu Pro Phe Tyr Ser Asn Val Ser 50 55 60 Trp Tyr Tyr Ser Leu Thr Thr Asn Asn Ala Ala Thr Lys Arg Thr Asp 65 70 75 80 Asn Pro Ile Leu Asp Phe Lys Asp Gly Ile Tyr Phe Ala Ala Thr Glu 85 90 95 His Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Asn 100 105 110 Thr Ser Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Ile Ile 115 120 125 Lys Val Cys Asn Phe Asp Phe Cys Tyr Asp Pro Tyr Leu Ser Gly Tyr 130 135 140 Tyr His Asn Asn Lys Thr Trp Ser Ile Arg Glu Phe Ala Val Tyr Ser 145 150 155 160 Ser Tyr Ala Asn Cys Thr Phe Glu Tyr Val Ser Lys Ser Phe Met Leu 165 170 175 Asn Ile Ser Gly Asn Gly Gly Leu Phe Asn Thr Leu Arg Glu Phe Val 180 185 190 Phe Arg Asn Val Asp Gly His Phe Lys Ile Tyr Ser Lys Phe Thr Pro 195 200 205 Val Asn Leu Asn Arg Gly Leu Pro Thr Gly Leu Ser Val Leu Gln Pro 210 215 220 Leu Val Glu Leu Pro Val Ser Ile Asn Ile Thr Lys Phe Arg Thr Leu 225 230 235 240 Leu Thr Ile His Arg Gly Asp Pro Met Pro Asn Asn Gly Trp Thr Ala 245 250 255 Phe Ser Ala Ala Tyr Phe Val Gly Tyr Leu Lys Pro Arg Thr Phe Met 260 265 270 Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys Ala 275 280 285 Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Leu Thr Val 290 295 300 Gln Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr Gln 305 310 315 320 Ser Val Val Arg Phe Pro Asn Ile Thr Asn Val Cys Pro Phe His Lys 325 330 335 Val Phe Asn Ala Thr Arg Phe Pro Ser Val Tyr Ala Trp Glu Arg Thr 340 345 350 Lys Ile Ser Asp Cys Ile Ala Asp Tyr Thr Val Phe Tyr Asn Ser Thr 355 360 365 Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Ser Lys Leu Ile 370 375 380 Asp Leu Cys Phe Thr Ser Val Tyr Ala Asp Thr Phe Leu Ile Arg Phe 385 390 395 400 Ser Glu Val Arg Gln Val Ala Pro Gly Gln Thr Gly Val Ile Ala Asp 405 410 415 Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp 420 425 430 Asn Thr Ala Lys Gln Asp Val Gly Asn Tyr Phe Tyr Arg Ser His Arg 435 440 445 Ser Thr Lys Leu Lys Pro Phe Glu Arg Asp Leu Ser Ser Asp Glu Asn 450 455 460 Gly Val Arg Thr Leu Ser Thr Tyr Asp Phe Asn Pro Asn Val Pro Leu 465 470 475 480 Glu Tyr Gln Ala Thr Arg Val Val Val Leu Ser Phe Glu Leu Leu Asn 485 490 495 Ala Pro Ala Thr Val Cys Gly Pro Lys Leu Ser Thr Gln Leu Val Lys 500 505 510 Asn Gln Cys Val Asn Phe Asn Phe Asn Gly Leu Lys Gly Thr Gly Val 515 520 525 Leu Thr Asp Ser Ser Lys Arg Phe Gln Ser Phe Gln Gln Phe Gly Lys 530 535 540 Asp Ala Ser Asp Phe Ile Asp Ser Val Arg Asp Pro Gln Thr Leu Glu 545 550 555 560 Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr 565 570 575 Pro Gly Thr Asn Thr Ser Leu Glu Val Ala Val Leu Tyr Gln Asp Val 580 585 590 Asn Cys Thr Asp Val Pro Thr Thr Ile His Ala Asp Gln Leu Thr Pro 595 600 605 Ala Trp Arg Ile Tyr Ala Thr Gly Thr Asn Val Phe Gln Thr Gln Ala 610 615 620 Gly Cys Leu Ile Gly Ala Glu His Val Asn Ala Ser Tyr Glu Cys Asp 625 630 635 640 Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr His Thr Ala Ser Ile 645 650 655 Leu Arg Ser Thr Ser Gln Lys Ala Ile Val Ala Tyr Thr Met Ser Leu 660 665 670 Gly Ala Glu Asn Ser Ile Ala Tyr Ala Asn Asn Ser Ile Ala Ile Pro 675 680 685 Thr Asn Phe Ser Ile Ser Val Thr Thr Glu Val Met Pro Val Ser Met 690 695 700 Ala Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Ile 705 710 715 720 Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu 725 730 735 Asn Arg Ala Leu Ser Gly Ile Ala Ile Glu Gln Asp Lys Asn Thr Gln 740 745 750 Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys 755 760 765 Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys 770 775 780 Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr 785 790 795 800 Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Gly 805 810 815 Ile Ser Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr 820 825 830 Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Ala Tyr Thr Ala 835 840 845 Ala Leu Ile Ser Gly Thr Ala Thr Ala Gly Trp Thr Phe Gly Ala Gly 850 855 860 Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn 865 870 875 880 Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile 885 890 895 Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Glu Ser Leu Thr 900 905 910 Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn 915 920 925 Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly 930 935 940 Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val 945 950 955 960 Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser 965 970 975 Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg 980 985 990 Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 995 1000 1005 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1010 1015 1020 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1025 1030 1035 Thr Tyr Ile Pro Ser Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1040 1045 1050 Ile Cys His Glu Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1055 1060 1065 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1070 1075 1080 Glu Pro Lys Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1085 1090 1095 Cys Asp Val Val Ile Gly Ile Ile Asn Asn Thr Val Tyr Asp Pro 1100 1105 1110 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1115 1120 1125 Phe Lys Asn His Thr Ser Pro Asp Ile Asp Leu Gly Asp Ile Ser 1130 1135 1140 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1145 1150 1155 Leu Asn Glu Val Ala Arg Asn Leu Asn Glu Ser Leu Ile Asp Leu 1160 1165 1170 Gln Glu Leu Gly Tyr Glu Gln Gly Ile High Pro High Tyr 1175 1180 1185 Val Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1190 1195 1200 Thr Ile Leu Leu Cys Met Thr Ser Cys Ser Cys Leu Lys 1205 1210 1215 Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp 1220 1225 1230 Ser Glu Pro Val Leu Gly Val Val Leu His Tyr Thr 1235 1240 1245 <210> 5 <211> 1273 <212> PRT <213> SARS-CoV-2 <400> 5 Met Phe Val Phe Leu Val Leu Pro Leu Val Ser Ser Gln Cys Val 1 5 10 15 Asn Leu Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe 20 25 30 Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu 35 40 45 His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp 50 55 60 Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp 65 70 75 80 Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu 85 90 95 Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser 100 105 110 Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile 115 120 125 Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr 130 135 140 Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr 145 150 155 160 Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu 165 170 175 Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe 180 185 190 Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr 195 200 205 Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu 210 215 220 Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr 225 230 235 240 Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser 245 250 255 Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro 260 265 270 Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala 275 280 285 Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys 290 295 300 Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val 305 310 315 320 Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys 325 330 335 Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala 340 345 350 Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu 355 360 365 Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro 370 375 380 Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe 385 390 395 400 Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly 405 410 415 Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys 420 425 430 Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn 435 440 445 Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe 450 455 460 Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys 465 470 475 480 Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly 485 490 495 Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val 500 505 510 Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys 515 520 525 Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn 530 535 540 Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu 545 550 555 560 Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val 565 570 575 Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe 580 585 590 Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val 595 600 605 Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile 610 615 620 His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser 625 630 635 640 Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val 645 650 655 Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala 660 665 670 Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala 675 680 685 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 690 695 700 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 705 710 715 720 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 725 730 735 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 740 745 750 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 755 760 765 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 770 775 780 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 785 790 795 800 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 805 810 815 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 820 825 830 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 835 840 845 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 850 855 860 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 865 870 875 880 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 885 890 895 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 900 905 910 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 915 920 925 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 930 935 940 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 945 950 955 960 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 965 970 975 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 980 985 990 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 995 1000 1005 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn 1010 1015 1020 Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys 1025 1030 1035 Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro 1040 1045 1050 Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val 1055 1060 1065 Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His 1070 1075 1080 Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn 1085 1090 1095 Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln 1100 1105 1110 Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val 1115 1120 1125 Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro 1130 1135 1140 Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn 1145 1150 1155 His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn 1160 1165 1170 Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu 1175 1180 1185 Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu 1190 1195 1200 Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu 1205 1210 1215 Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met 1220 1225 1230 Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys 1235 1240 1245 Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro 1250 1255 1260 Val Leu Lys Gly Val Lys Leu His Tyr Thr 1265 1270

Claims

1. A vaccine for preventing β-CoV infection, comprising: A vaccine comprising at least one viral vector comprising a β-CoV DNA sequence encoding the S protein of β-CoV.

2. The vaccine of claim 1 , wherein the vector is an adenovirus vector.

3. The vaccine of claim 2 , wherein the vector is a fully deleted adenoviral vector that does not contain any endogenous genes.

4. The vaccine of any one of claims 1 to 3, wherein the β-CoV DNA sequence is a SARS-2 β-CoV DNA sequence.

5. 5. The vaccine of claim 4, wherein the SARS-2β-CoV DNA sequence is the entire sequence encoding the S protein.

6. The vaccine of claim 4, wherein the SARS-2β-CoV DNA sequence is a partial sequence encoding the S protein.

7. The vaccine of claim 4, wherein the SARS-2β-CoV DNA sequence is a partial sequence encoding an S protein from which the receptor binding domain has been deleted.

8. 5. The vaccine of claim 4, wherein the SARS-2β-CoV DNA sequence is a subsequence encoding an S protein, in which the receptor binding domain sequence is replaced by DNA encoding a peptide linker.

9. The vaccine of any one of claims 1 to 8, further comprising a packaging plasmid based on an adenovirus selected from the group consisting of serotypes Ad2, Ad5, Ad6 and Ad35, and combinations thereof.

10. The vaccine of any one of claims 1 to 9, wherein at least one viral vector is contained in a packaging cell.

11. 11. The vaccine of claim 10, wherein the packaging cells are encapsidated into a capsid selected from the group consisting of serotypes Ad2, Ad5, Ad6 and Ad35, and combinations thereof.

12. 12. The vaccine of any of claims 1 to 11, wherein the β-CoV DNA sequence is a SARS-2 β-CoV DNA sequence, and the viral vector comprises at least a second β-CoV DNA sequence from a SARSr virus, the second β-CoV DNA sequence encoding the S protein of the SARSr virus.

13. At least one viral vector comprising a SARS-2β-CoV DNA sequence encoding the S protein of SARS-2β-CoV; at least one packaging plasmid based on an adenovirus selected from the group consisting of serotypes Ad2, Ad5, Ad6 and Ad36, and combinations thereof; A vaccine for preventing SARS-2 infection comprising: At least one viral vector and at least one packaging plasmid are contained in a packaging cell; A vaccine wherein the packaging cells are encapsidated into capsids selected from the group consisting of serotypes Ad2, Ad5, Ad6 and Ad35, and combinations thereof.

14. 14. The vaccine of claim 13, wherein the SARS-2 β-CoV DNA sequence encodes a partial S protein of the SARS-2 virus.

15. A vaccine for preventing β-CoV infection, comprising: A vaccine comprising at least one β-CoV RNA sequence encoding the S protein of β-CoV.

16. The vaccine of claim 18, wherein the RNA is mRNA.

17. The vaccine of any of claims 15 to 16, wherein the β-CoV RNA sequence is a SARS-2 β-RNA sequence.

18. The vaccine of any one of claims 15 to 17, wherein the SARS-2β-CoV RNA sequence is the entire sequence encoding the S protein.

19. The vaccine of any one of claims 15 to 16, wherein the SARS-2β-CoV RNA sequence is a partial sequence encoding the S protein.

20. 20. The vaccine of claim 19, wherein the SARS-2β-CoV RNA sequence is a subsequence encoding an S protein from which the receptor binding domain has been deleted.

21. 20. The vaccine of claim 19, wherein the SARS-2β-CoV RNA sequence is a subsequence encoding the S protein, in which the receptor binding domain sequence is replaced by RNA encoding a peptide linker.

22. The vaccine of any one of claims 15 to 21, further comprising an expression vector that delivers the genetic information of β-CoV RNA.

23. 20. The vaccine of claim 18, wherein the expression vector is an engineered viral vector.

24. A vaccine for preventing β-CoV infection, comprising: A vaccine comprising at least one β-CoV protein sequence encoding the S protein of β-CoV.

25. 25. The vaccine of any of claims 24, wherein the β-CoV RNA sequence is a SARS-2 β-CoV protein sequence.

26. The vaccine of any of claims 24 to 25, wherein the SARS-2β-CoV protein sequence is the entire sequence encoding the S protein.

27. The vaccine of any one of claims 24 to 25, wherein the SARS-2β-CoV protein sequence is a partial sequence encoding the S protein.

28. 28. The vaccine of claim 27, wherein the SARS-2 β-CoV protein sequence is a partial S protein sequence in which the receptor binding domain has been removed.

29. 28. The vaccine of claim 27, wherein the SARS-2 β-CoV protein sequence is a partial S protein sequence, in which the receptor binding domain sequence is replaced by a peptide linker.

30. 1. A method of vaccinating a mammalian subject against infection from at least one group of β-CoVs, comprising: Classifying the broad group of β-CoVs into homology groups based on similarities in the β-CoV RNA sequences that encode their S proteins; identifying at least one consensus sequence for each homology group that has greater than 60% sequence identity to all other members of the homology group; Preparing a viral vector comprising at least a portion of a consensus sequence from at least one homology group; A method comprising:

31. 31. The method of claim 30, wherein the consensus sequence is selected from the group consisting of a DNA sequence, an RNA sequence, a protein sequence, and combinations thereof.

32. 31. The method of claim 30, wherein preparing a viral vector comprises including at least a portion of a consensus sequence from two or more homology groups.

33. 31. The method of claim 30, further comprising injecting the vaccine into a mammalian subject.

34. 1. A method of vaccinating a mammalian subject against infection from at least one group of β-CoVs, comprising: Classifying the broad group of β-CoVs into homology groups based on similarities in the β-CoV DNA, RNA or protein sequences encoding their S proteins; identifying at least a portion of a β-CoV protein sequence for each homology group that has greater than 60% sequence identity to all other members of the homology group; preparing a DNA, RNA or protein vaccine comprising at least a portion of a β-CoV protein sequence from at least one homology group; A method comprising:

35. 35. The method of claim 34, further comprising injecting the vaccine into a mammalian subject.

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