KR20220039078A - Vaccine composition for preventing or treating infection of SARS-CoV-2 comprising a modified Spike protein of SARS-CoV-2 - Google Patents

Abstract

The present invention provides a recombinant antigen protein, and a vaccine composition comprising the same, wherein the recombinant antigen protein is derived from a SARS-coronavirus-2 spike protein, in which all or a part of the S2 subunit of the spike protein is removed, and the recombinant antigen protein is a SARS-coronavirus-2 infection prevention or treatment antigen.

Description

SARS-coronavirus-2 vaccine composition for preventing or treating infection, comprising a modified spike protein of SARS-CoV-2 comprising a modified Spike protein of SARS-CoV -2}

The present invention relates to a vaccine composition for preventing SARS-coronavirus-2 (SARS-CoV-2) infection, and more particularly, SARS-coronavirus using an antigen derived from SARS-CoV-2 (SARS-CoV-2). It relates to a vaccine composition for preventing virus-2 infection.

SARS-CoV-2 (SARS-CoV-2) is called Severe Acute Respiratory Syndrome Coronavirus 2 or Covid 19 (COVID19), and is named Corona 19 in Korea. SARS-coronavirus-2 was first discovered at the Huanan Fish Market in Wuhan on December 12, 2019. It is an RNA virus, and a human-to-human infection was confirmed.

SARS-coronavirus-2 is a virus that requires handling at a biosafety level 3 research facility (BSL-3 facility), and the virus's reproduction index (R0) is estimated to be 1.4 to 3.9. This means that one patient can transmit the virus to a minimum of 1.4 to a maximum of 3.9 people, that is, it is estimated that the control of the infectious disease caused by SARS-coronavirus-2 is quite difficult, and as of March 31, 2020, worldwide There were 785,867 infections and 37,827 deaths.

Symptoms such as fever, shortness of breath, kidney and liver damage, cough, and pneumonia are observed for 2 to 14 days after infection with the virus, and a therapeutic agent has not yet been developed.

In a situation where no treatment has been developed, research on vaccines is urgently needed to prevent infection and spread to the community. Since the epidemic virus is usually a high-risk pathogen, inactivated and live vaccines have a high risk in the production and human administration of vaccine substances. In particular, in the case of a live vaccine, it takes a very long time to attenuate and prove safety. The inventors of the present invention completed the present invention by studying a recombinant protein vaccine applicable to the current pandemic novel infectious disease in terms of versatility, safety, efficacy, and commercialization.

1. Zhou Z, Post P, Chubet R, et al. A recombinant baculovirus-expressed S glycoprotein vaccine elicits high titers of SARS-associated coronavirus (SARS-CoV) neutralizing antibodies in mice. Vaccine. 2006;24(17):3624-3631. 2. Dai L, Zheng T, Xu K, et al. A Universal Design of Betacoronavirus Vaccines against COVID-19, MERS, and SARS. Cell. 2020;182(3):722-733.e11.

Therefore, the problem to be solved by the present invention is to provide a novel antigen for preventing SARS-coronavirus-2 infection, a vaccine composition comprising the antigen, or a method for manufacturing the same in order to solve the above problems. The present invention is to provide a recombinant protein vaccine, a method for preventing SARS-coronavirus-2 infection using the same, or a use of the recombinant protein vaccine for preventing SARS-coronavirus-2 infection.

In order to solve the above problems, one aspect of the present invention provides a recombinant antigen protein for preventing SARS-coronavirus-2 (SARS-CoV-2) infection, and a vaccine composition comprising the same.

The inventors of the present invention found that the recombinant antigen produced by cleavage of a specific site of the S1 and/or S2 domain of the SARS-coronavirus-2 spike protein (S protein) has significantly higher ADE (Antibody-Dependent Enhancement) side effects. The present invention was completed on the basis of the reduction.

The present invention includes a recombinant antigen protein derived from SARS-coronavirus-2 spike protein (S protein), in which all or part of the S2 subunit of the spike protein has been removed, wherein the recombinant antigen protein is SARS- Provided is a recombinant antigen protein that is an antigen for preventing or treating coronavirus-2 infection. It is known that SARS-CoV-2 interacts with the receptor of the host cell through the S1 subunit of the spike protein, and the virus penetrates into the cell as the membrane fuses through the S2 subunit. there is. The RBD (Receptor-Binding Domain, located at 331-524 of the full-length amino acid sequence) of the spike protein of SARS-CoV-2 is included in the S1 subunit. After the S protein is transcribed to full-length, it forms a two-subunit complex of S1/S2 through post-translational modification or subsequent protease cleavage, and the S1 subunit is angiotensin-converting enzyme 2 (Angiotensin-converting enzyme 2, It contains a receptor binding domain (RBD) that binds to ACE 2), and the S2 subunit includes a transmembrane domain and a cytosolic domain that penetrate the stem and cytoplasm. The inventors of the present invention confirmed that when all or part of the S2 subunit of the SARS-CoV-2 spike protein was removed, the shape maintenance of the antigen protein and the increase in the antigen protein expression level were achieved and completed the present invention. became Since the spike protein is a membrane protein, it cannot be expressed soluble, and the amount of soluble expression of the ectodomain excluding the transmembrane domain and cytosolic domain is extremely limited. Since the S1 subunit and the S2 subunit are independent domains, the critical neutralizing epitope is located in the RBD part included in the S1 subunit. Therefore, when only the S1 part excluding S2 is expressed, the soluble expression level is maintained while maintaining the critical neutralizing epitope. can increase If you look at the structure of the SARS coronavirus spike protein or the structure of the SARS coronavirus 2 spike protein (PDB ID: 6VSB), even if the 685/686 amino acid position is cut by furin protease and separated into S1 and S2 subunits, the N-terminal of the S2 subunit The part and the C-terminal part of the S1 subunit are still interacting, so it is expected that the C-terminal structure of the S1 subunit can be stabilized if some of the N-terminal amino acids of the S2 subunit are left. In addition, the recombinant antigen protein having a specific sequence of the present invention can obtain an improved expression level, an excellent protein stabilization effect, or a neutralizing antibody induction effect.

The term “all” removal of the S2 subunit means that the S2 subunit is completely removed from the S protein and the signal peptide of the S protein (present at the 1st to 13th amino acid positions of the full-length S peptide sequence) and the S1 subunit connected thereto It is understood to mean that only a unit (present at amino acid positions 14 to 685 of the full-length S peptide sequence) is present. The removal of “part” of the S2 subunit means that some sequence of the S2 subunit peptide of the S protein remains at the C terminus of the S1 subunit, preferably starting at position 686 of the S protein. means that at least 26 peptide sequences of the S2 subunit are left at the C-terminus of the S1 subunit, more preferably at least 16, more preferably at least 11 peptide sequences are left.

In one aspect of the present invention, the recombinant antigen protein has the entire S2 subunit removed from the SARS-coronavirus-2 spike protein, and preferably has an amino acid sequence represented by SEQ ID NO: 1. The C-terminal portion of the S1 subunit does not form a stable structure and is a freely moving portion exposed to a solution. It is not seen in the cryoEM structure, and removing these portions does not affect the structure of S1. It is also possible to obtain the form of the S1 antigen. In another embodiment, in the recombinant antigen protein, four amino acids corresponding to positions 683 to 686 of the full-length S protein may be further removed, which is referred to as SK-S1Δ in the specification, and may be represented by SEQ ID NO: 2 there is. The four amino acids corresponding to positions 683 to 686 are protease cleavage sites. When linking a part that can help enhance immunity, such as the Tetanus P2 domain, to the C terminus of the S1 subunit, removing the protease cleavage site and linking can prevent release of P2 peptide during expression and purification.

The N-terminal domain of S1 has a structure independent of the RBD domain, and in the case of the SARS coronavirus spike protein, this part contains 3 out of 5 immunodominant regions of the spike protein. known not to do so. Therefore, if this part is removed, the immunodominant region is reduced, so that the RBD domain can induce more antibodies, lower the possibility of potential ADE, and increase the protein expression level can be expected. In one embodiment of the present invention, the recombinant antigen protein is a SARS-coronavirus-2 spike protein in which a part of the N-terminal domain of the S1 subunit and a part of the S2 subunit have been removed. In another embodiment, the recombinant antigen protein may be linked to its own signal peptide or a heterologous signal peptide. Preferably, the cleaved and removed position in a part of the S1 subunit may be amino acid position 295, preferably position 293, more preferably position 291, more preferably position 289, starting from the N-terminus of the S protein. . The recombinant antigen protein has a part of the S1 subunit and a part of the amino acid sequence of the S2 subunit, and the amino acid sequence of the human albumin signal peptide of SEQ ID NO: 19 in the amino acid sequence at positions 290 to 701 of the S protein is at the N-terminus. can be connected The recombinant antigen protein is represented by SEQ ID NO: 6.

As used herein, the term "recombinant antigen protein" is an antigen for the prevention or treatment of SARS-CoV-2 infection, specifically, the amino acid sequence of a specific section selected at a specific position of the SARS-CoV-2 spike protein. protein containing. The recombinant antigen protein refers to a protein artificially created through cleavage of a partial region of the SARS-CoV-2 spike protein and binding to a foreign gene.

In another embodiment, the C-terminus of the recombinant antigen protein may optionally further include a T cell epitope, and any T cell epitope domain may be used without limitation. Preferably, one of the T cell epitopes may include a Tetanus Toxoid Epitope P2 domain (SEQ ID NO: 20). The P2 domain may be bound to exhibit a more improved immune enhancing effect. Preferably, the P2 domain may be located at the C-terminus of the recombinant protein antigen. The linker is, for example, no more than 16 amino acids in length and may preferably consist of no more than 6 amino acids. The amino acid used in the linker is at least one of G (Gly, glycerin), S (Ser, serine), and A (Ala, alanine), preferably Gly-Ser-Gly-Ser-Gly (GSGSG), Gly-Ser -Ser-Gly (GSSG), Gly-Ser-Gly-Gly-Ser (GSGGS), Gly-Ser-Gly-Ser (GSGS), and Gly-Ser-Gly-Ser-Ser-Gly (GSGSSG) It may be any one or more peptide linkers selected from, preferably, a GSGSG peptide linker for the purpose of the present invention.

The recombinant antigen protein according to a specific embodiment of the present invention consists of any one or more amino acid sequences selected from the group consisting of the amino acid sequences of SEQ ID NOs: 1 to 6. The recombinant antigenic protein induces an excellent cellular immune response.

The present invention provides a nucleotide sequence encoding a recombinant protein antigen as defined above, and provides a construct for antigen expression for recombinant protein antigen expression. One embodiment of the present invention provides an expression construct for the production of a recombinant antigen protein for the prevention or treatment of SARS-coronavirus-2 infection. In one embodiment, SARS-coronavirus-2 comprising a nucleic acid sequence encoding a peptide from which all or part of the S2 subunit of the spike protein (S protein) has been removed - a recombinant antigen protein for preventing or treating coronavirus-2 infection An expression construct for production is provided. The expression construct is a nucleic acid sequence encoding a peptide from which all or part of the S2 subunit of the SARS-coronavirus-2 spike protein (S protein) has been removed, and a nucleic acid sequence encoding a signal peptide of its own or heterologous origin is added. can be fused. The expression construct may further be fused with a nucleic acid sequence encoding the P2 domain of Tetanus toxin.

As used herein, the term “expression construct” is understood to mean a nucleic acid molecule containing only a minimal element for protein expression in a cell.

Preferably, it provides any one or more nucleotide sequences selected from the group consisting of SEQ ID NOs: 7 to 12, which can obtain excellent recombinant antigen proteins in the baculovirus expression system (BEVS). Or, the expression construct contains any one or more nucleotide sequences selected from the group consisting of SEQ ID NOs: 13 to 18, which can obtain excellent recombinant antigen proteins in a Chinese Hamster ovary cell (CHO) mammalian expression system. It is possible to provide an expression construct that Preferably said nucleotide sequence is a DNA sequence.

The antigen expression construct of one embodiment is a nucleic acid sequence encoding a peptide in which all or part of the S2 subunit of the SARS-coronavirus-2 spike protein (S protein) has been removed from the nucleotide sequence encoding its own or heterologous signal sequence may be provided in connection with As used herein, the term "signal peptide" or "signal sequence" is used interchangeably herein and refers to a short peptide present at the N-terminus of a newly synthesized polypeptide chain (generally 5-30 amino acids in length, but is not limited thereto). The signal peptides referred to herein are eliminated during protein secretion. In one embodiment, the recombinant antigen protein may be linked to the N-terminus of a receptor binding domain in which a heterologous signal sequence is extended. The 'self signal sequence' refers to the signal sequence of the spike protein of ARS-CoV-2. The 'heterologous signal sequence' refers to an externally introduced signal sequence, not the signal sequence of the SARS-CoV-2 spike protein. The signal sequence can be used by replacing it with a signal peptide sequence. Preferred heterologous signal sequences include a murine phosphatase signal peptide sequence, a honeybee melittin signal peptide sequence, a human albumin signal peptide sequence, and the like, and the amino acid sequence of the human albumin signal peptide may be represented by SEQ ID NO: 19.

Preferably, it may include any one or more nucleotide sequences selected from the group consisting of nucleotide sequences of SEQ ID NOs: 7 to 12, which can obtain excellent recombinant antigen proteins in a baculovirus expression system. Or preferably, it may include any one or more nucleotide sequences selected from the group consisting of SEQ ID NOs: 13 to 18 in an expression system using a Chinese hamster ovary (CHO) cell as a host cell. Preferably said nucleotide sequence is a DNA sequence.

Recombinant antigen proteins of the invention can be produced by cloning and expression in prokaryotic or eukaryotic expression systems using suitable expression vectors. Any method known in the art may be used. Preferably, in consideration of the purpose of the present invention and protein expression rate, BEVS, CHO or E. coli expression systems may be used, and BEVS and/or CHO expression systems may be preferably used. A vector may be of any suitable type and may include, but is not limited to, phage, virus, plasmid, phagemid, cosmid, bacmid, and the like. For example, a DNA molecule encoding an antigen of the present invention is inserted into an appropriately constructed expression vector by techniques well known in the art. Known techniques are described in Zhou Z, Post P, Chubet R, et al. A recombinant baculovirus-expressed S glycoprotein vaccine elicits high titers of SARS-associated coronavirus (SARS-CoV) neutralizing antibodies in mice. Vaccine. 2006;24(17):3624-3631. doi:10.1016/j.vaccine.2006.01.059 (BaculoSystems), Dai L, Zheng T, Xu K, et al. A Universal Design of Betacoronavirus Vaccines against COVID-19, MERS, and SARS. Cell. 2020;182(3):722-733.e11. doi:10.1016/j.cell.2020.06.035 (CHO system) and the like can be referred to.

The expression construct according to an embodiment of the present invention uses a baculovirus expression system (BEVS).

The baculovirus expression system can be used without limitation, which is already widely used for recombinant protein production in the industry. For example, commercially available baculovirus vectors such as pBAC4x-1 (Novagen) can be used. Suitable baculovirus promoters for use in the present invention are well known in the literature. As the baculovirus promoter, a commonly used promoter such as polyhedrin and p10 promoter may be used. Also provided are a recombinant Bacmid obtained by transforming Escherichia coli with a baculovirus vector containing an expression construct comprising a nucleotide sequence encoding the antigenic protein, and a recombinant baculovirus comprising the same as a genome. Host cells containing the recombinant baculovirus or transfected with the recombinant baculovirus are also included in the scope of the present invention.

DNA molecules comprising a nucleotide sequence encoding an antigenic protein of the present invention can be inserted into vectors having transcriptional and translational control signals. Cells stably transformed with the introduced DNA can also be selected by introducing one or more markers that allow selection of host cells containing the expression vector. The marker may provide, for example, antibiotic resistance, a deficient nutrient synthesis gene, and the like. Once a vector or DNA sequence containing the construct has been prepared for expression, the DNA construct may be subjected to any of a variety of suitable means, i.e., transformation, transfection, conjugation, protoplast fusion, electroporation, calcium phosphate-precipitation, It can be introduced into suitable host cells by direct microinjection or the like.

Preferred host cells are eukaryotic host cells, for example, Spodopterafrugiperda (Sf) cells such as Sf9 and Sf21 using the Baculovirus expression system as insect cells, Trichoplusia ni cells such as Hi-5 cells, and Drosophila S2 cells. and the mammalian cells may include Chinese Hamster Ovary (CHO) cells. A suitable host cell line may be any Chinese Hamster Ovary (CHO) cell line. The term 'host cell' refers to a cell capable of growing in culture and expressing a desired protein recombinant product protein. Suitable cell lines may include, but are not limited to, for example, CHO K1, CHO pro3-, CHO DG44, CHO P12, and the like.

It is possible to obtain a recombinant antigen protein having an excellent expression rate through the host cell. As non-limiting examples, examples of the eukaryotic host cells may include yeast, algae, plants, C. elegans (or nematodes), etc. within the scope that does not impair the object of the present invention, and prokaryotic host cells are, for example, E. coli (E. coli, B. subtilis), Salmonella typhi, and may contain bacterial cells such as mycobacteria. After introduction of the vector, the host cells are propagated in a normal medium or a selective medium (selected for growth of vector-containing cells). As a result of expression of the cloned gene sequence(s), the desired protein is produced. Purification of the recombinant antigenic protein can be carried out by any conventional procedure involving any one of methods known for this purpose, namely extraction, precipitation, chromatography, electrophoresis, and the like.

Another aspect of the present invention provides a method for producing the recombinant antigenic protein, the method may include culturing a host cell transformed with a vector containing the nucleotide sequence of the present invention and isolating the desired product. .

Another embodiment of the present invention provides a novel use of the recombinant protein antigen for preventing or treating SARS-coronavirus-2 infection, and administering the antigen to a subject to prevent or treat SARS-coronavirus-2 infection A method of preventing SARS-coronavirus-2 infection is provided.

In another embodiment of the present invention, the SARS-coronavirus-2 infection prevention or treatment vaccine composition comprising the recombinant antigen protein according to the present invention as an active ingredient is provided. The 'SARS-coronavirus-2 infection' is a concept that broadly includes not only the infection of the SARS-coronavirus-2 itself, but also various conditions (eg, respiratory disease, pneumonia, etc.) resulting from the infection of the virus. can be understood as In the present invention, the vaccine may be prepared by a conventional method well known in the art, and may optionally further include various additives that can be used in the preparation of a vaccine in the art. The vaccine composition according to the present invention may include the recombinant antigen protein and a pharmaceutically acceptable carrier. Although not limited thereto, for example, lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose as commonly used in formulations. , polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. The pharmaceutical composition of the present invention, in addition to the above components, TWEEN ™, non-ionic surfactants such as polyethylene glycol (PEG), antioxidants including ascorbic acid, lubricants, wetting agents, sweetening agents, flavoring agents, emulsifying agents, suspending agents, It may be used by further including a preservative and the like. In the present invention, the vaccine is prepared in unit dose form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person of ordinary skill in the art to which the present invention pertains. or it may be prepared by incorporation into a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer. A suitable dosage of the vaccine in the present invention may be prescribed variously depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity of the patient. can On the other hand, the dosage of the vaccine according to the present invention may be preferably 1 ~ 500 ug per dose. In one embodiment of the present invention, the vaccine comprising the recombinant antigen protein as an active ingredient may be administered into the body by intravenous injection, intramuscular injection, subcutaneous injection, transdermal delivery or airway inhalation, but is not limited thereto.

The vaccine composition may further include an immunological adjuvant, in order to enhance the immune response effect, further adding the nucleocapsid (N) protein of SARS-coronavirus-2 with or without the immunological adjuvant. may include

The immunological adjuvant is, for example, squalene (MF59), liposomes, TLR agonist, monophosphoryl lipid A (MPL) (AS04), magnesium hydroxide, magnesium carbonate hydroxide pentahydrate date, titanium didoxide, calcium selected from carbonate, barium oxide, barium hydroxide, barium peroxide, barium sulfate, calcium sulfate, calcium pyrophosphate, magnesium carbonate, magnesium oxide, aluminum hydroxide, aluminum phosphate and hydrated aluminum potassium sulfate (Alum) It may be any one or more, but is not limited thereto.

The 'SARS-coronavirus-2 nucleocapsid (N) protein' is an artificially made SARS-coronavirus-2 nucleocapsid (N) protein of SEQ ID NO: 21. The N protein can induce cellular immunity, and can be used together with the recombinant antigen protein obtained according to an embodiment of the present invention to induce increased protective immunogenicity. The construct for N protein expression of the protein of SEQ ID NO: 21 may be provided by fusion of a nucleotide sequence capable of expressing a human albumin signal peptide to the N-terminus of the N protein. Preferably, the nucleotide sequence optimized in the BEV expression system is represented by SEQ ID NO: 22, and the nucleotide sequence optimized in the CHO expression system is represented by SEQ ID NO: 23.

Optionally, the vaccine composition may further comprise an M protein of SARS-coronavirus-2.

The recombinant protein and/or recombinant virus vaccine according to an embodiment of the present invention has high safety.

The vaccine according to one embodiment of the present invention has excellent immunogenicity and has excellent efficacy as a vaccine.

The vaccine of the present invention is excellent in inducing neutralizing antibody.

The vaccine of the present invention is excellent in inducing cellular immunity.

The present invention is excellent in preventing or treating SARS-coronavirus-2 infection.

The recombinant antigen protein of the present invention can maintain a stable three-dimensional antigen protein structure. The recombinant antigen of the present invention can have high antibody inducibility. The recombinant protein of the present invention has a high expression rate. The recombinant antigen of the present invention does not cause the possibility of ADE (Antibody-Dependent Enhancement) side effects.

The following drawings attached to this specification illustrate preferred embodiments of the present invention, and serve to further understand the technical spirit of the present invention together with the above-described content of the invention, so the present invention is limited to the matters described in those drawings It should not be construed as being limited.
1 shows a schematic diagram of the structure of the SARS-CoV2 spike full-length protein domain.
Figure 2 is a schematic diagram of a construct for expression of a recombinant antigen protein made on the basis of the peptide sequence of the S protein.
3 to 14 show SEQ ID NOs: 7 to 18, respectively. 3 to 14, blue was used to indicate a signal peptide, orange was a peptide linker, and light green was used to indicate a nucleotide sequence encoding a Tetanus Toxoid Epitope P2 domain.

Hereinafter, examples and the like will be described in detail to help the understanding of the present invention. However, the embodiments according to the present invention may be modified in various other forms, and the scope of the present invention should not be construed as being limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art to which the present invention pertains.

1. Preparation of a construct for antigen expression using the SARS-coronavirus-2 Spike protein

S gene, N gene, and M gene sequences were prepared by referring to the sequence of Genbank # MN908947 Severe acute respiratory syndrome coronavirus 2 isolate Wuhan-Hu-1 to prepare the antigen protein used for vaccine production.

1 shows a schematic diagram of the structure of the SARS-CoV2 spike full-length protein domain. 2 shows a construct for expression of a recombinant antigen protein prepared based on the peptide sequence of the S protein.

Table 1 below shows the characteristics of the antigen protein obtained by the expression construct shown in FIG.

residues Length (aa) MW (kDa) PI (Isoelectric Point) Charge at pH7 Extinction Coefficient number of cys Cyc% S1 truncated 2 (SEQ ID NO: 5) 290~701 410 45.193 7.50 2.20 48665 15 3.7

SEQ ID NO: 5 is designated as SK-S1-T2, and in SEQ ID NO: 5, a part of the N-terminal domain of the S1 subunit of the spike protein is removed, and the N-terminal part of the S2 subunit is included at the C-terminus.

When each construct is expressed, an appropriate signal peptide can be added to each expression system so that the recombinant protein can be secreted into the periplasmic region or culture medium, or the original signal peptide can be replaced with a heterogeneous signal peptide. In the Spike protein, N-terminal 1-13 aa (MFVFLVLLPLVSS) is its own signal peptide, and it was replaced with human albumin signal peptide (SEQ ID NO: 19: MKWVTFISLLFLFSSAYS) in the baculovirus system, CHO cell expression system, and mammalian cell expression system.

The T cell epitope P2 domain was linked with a GSGSG peptide linker.

2. Preparation of constructs for antigen expression using other proteins

A construct for antigen expression was prepared based on the N protein gene. It was prepared by linking the human albumin signal peptide of SEQ ID NO: 19 to SEQ ID NO: 21.

3. Codon Optimization

The DNA sequence encoding the recombinant protein was synthesized with codons optimized for insect cells and CHO cells in GenScript, respectively.

The codon-optimized sequences for each expression system are as follows.

BEVS CHO designation SEQ ID NO: SEQ ID NO: SK-S1 7 13 SK-S1△ 8 14 SK-S1-P2 9 15 SK-S1△-P2 10 16 SK-S1-T2 11 17 SK-S1-T2-P2 12 18

4. Manufacture of Recombinant Protein Vaccines

The effect of the vaccine was confirmed by the following procedure using Baculo, and CHO.

1.1 Production of recombinant proteins

1.1.1 Recombinant protein production using baculovirus expression system

1.1.1.1 Each gene was synthesized to express the SARS-coronavirus-2 antigen gene. The construct for protein expression and the construct for N protein expression of FIG. 2 were prepared, respectively. Transfer vector pFastBac vector was cloned by inserting the prepared construct gene into the vector, and the gene sequence was analyzed.

1.1.1.2 The prepared plasmid was transformed into E. coli for bacmid production to prepare a recombinant bacmid, and the gene sequence was analyzed.

1.1.1.3 Recombinant Vacmid was inoculated into Sf9 cells cultured as a monolayer, transfected, and recombinant baculovirus (P0) was prepared and quantified by the plaque test.

1.1.1.4 The cultured Hi-5 cells were infected with the recombinant baculovirus to secure the P1 virus, and the antigen protein produced in the supernatant was confirmed.

1.1.1.5 The antigen protein produced by infecting the P1 virus with Hi-5 cells was recovered.

1.1.1.5 Recombinant Protein Purification

The collected recombinant protein was filtered using a filter, and the recombinant protein was purified using an appropriate chromatography method (Ion Exchange, Size Exclusion, etc.).

1.1.2 Recombinant protein production using CHO cell expression system

1.1.2.1 Each gene was synthesized to express the SARS-coronavirus-2 antigen gene. The synthesized gene was inserted into the expression vector and cloned, and the gene sequence was analyzed.

1.1.2.2 The recombinant plasmid was transformed into CHO cells for protein production (CHO K-1 cell line).

1.1.2.3 Transformed cells expressing the recombinant protein were identified using antibiotics.

1.1.2.4 The identified transformed CHO cells were mass-cultured and the recombinant protein was harvested.

1.1.2.5 Recombinant Protein Purification

The collected recombinant protein was filtered using a filter, and the recombinant protein was purified using an appropriate chromatography method (Ion Exchange, Size Exclusion, etc.).

1.1.5 Recombinant protein identification and quantification

1.1.5.1 Expression of the recombinant protein was confirmed using SDS-PAGE and Western blot.

1.1.5.2 Recombinant protein was quantified using basic total protein quantification methods (Lowry method, BCA method, etc.).

5. Evaluation of Recombinant Antigen Proteins

A. Animal testing

1.2.1 Immunogenicity Test

1.2.1.1 In an animal model, the purified recombinant protein was combined with an immune adjuvant (eg / aluminum hydroxide) and inoculated 2-3 times with an interval of 2-3 weeks.

1.2.1.2 Measure changes in body weight and body temperature to confirm safety

1.2.1.3 Two to three weeks after the final inoculation, whole blood was used to obtain serum and splenocytes isolated.

1.2.2 Protection Test

1.2.2.1 In an animal model, the purified recombinant protein was combined with an immune adjuvant (eg / aluminum hydroxide) and inoculated 2-3 times with an interval of 2-3 weeks.

1.2.2.2 Two to three weeks after the final inoculation, a lethal dose of wild-type SARS-coronavirus-2 virus was infected.

1.2.2.3 Virus shedding in the nasal cavity, respiratory tract, and organs was evaluated for one week after infection.

1.2.2.4 For 2 weeks after infection, changes in body weight and body temperature, mortality, etc. were evaluated.

B. Immunogenicity Assessment

1.3.1 IgG ELISA

1.3.1.1 Coating antigen (RBD, S1, S2, N, etc.) for coating on a 96-well-plate, and blocking the plate with a blocking buffer. The sample (serum) was reacted on the plate. An IgG detection antibody was reacted on the plate. A substrate buffer was added to develop color, and absorbance was measured.

1.3.2 Pseudovirus production

1.3.2.1 Cloning the S protein gene of SARS-coronavirus-2 into an expression vector. A reporter gene was cloned into a transfer vector. The two genes were transformed into pseudovirus-producing cells to prepare a pseudovirus expressing a reporter protein.

1.3.3 Neutralizing antibody evaluation

1.3.3.1 Passage-diluted sample (serum) is reacted with pseudovirus. The reacted pseudovirus was infected with cells for infection (Vero E6, etc.) cultured in a 96-well-plate and cultured. After 4-6 hours, it was washed with PBS and replaced with a fresh medium. The neutralizing antibody titer was evaluated by comparing the reporter protein expression level after culturing for 24 to 72 hours.

1.3.4 Cellular Immunity Assessment

1.3.4.1 96-well-plates were coated with anti-interferon-gamma antibody (anti-IFN-γ antibody). The plate was blocked with a blocking buffer, splenocytes and stimulator antigen (Stimulate) were added, and incubated for 24 to 36 hours. Interferon-gamma detection antibody was reacted, and a substrate was added to react. Immune cells were evaluated using an ELISPOT reader.

1.3.4.2 For immune characterization, the immune cell-specific antibody and cytokine antibody were separated and reacted with splenocytes for 2 hours. T cell distribution and cytokine expression rate were measured by flow cytometry.

C. Assessment of antigenicity of antigens for vaccines

BioLayer Interferometry (BLI) was used to evaluate binding to CR3022. CR3022 is a human monoclonal antibody against the recombinant SARS-CoV-2 Spike Glycoprotein S1. (CAT# of Abcam: ab273073)

BLI measures the affinity constant KD value (Kdis/Kon) through association and dissociation between the antibody and antigen, and the smaller the value, the higher the affinity. Corona 19 S-specific antibody was immobilized using Octet K2 on ProA sensor chip (ForteBio). Association was measured by dipping the sensor chip into an antigen sample diluted 2-fold from 100 nM, and dissociation was measured by dipping into a well containing only kinetic buffer. Using Octet Data Analysis software (11.0), the data obtained by subtracting the reference from the result value was analyzed by fitting the 1:1 binding model.

Through this, it was possible to secure synthetic sequences and information, protein expression confirmation, protein isolation and purification, and recombinant protein vaccine candidates.

Through this, sufficient antibodies and protective immunity can be induced to prevent corona infection.

number division note One peptide SK-S1 2 peptide SK-S1△ 3 peptide SK-S1-P2 4 peptide SK-S1△-P2 5 peptide SK-S1-T2 6 peptide SK-S1-T2-P2 7 nucleotides Optimized constructs for SK-S1 peptide expression in BEVS 8 nucleotides Optimized constructs for SK-S1Δ peptide expression in BEVS 9 nucleotides Optimized constructs for SK-S1-P2 peptide expression in BEVS 10 nucleotides Optimized constructs for expression of SK-S1Δ-P2 peptides in BEVS 11 nucleotides Optimized constructs for SK-S1-T2 peptide expression in BEVS 12 nucleotides Optimized constructs for expression of SK-S1-T2-P2 peptides in BEVS 13 nucleotides Optimized constructs for SK-S1 peptide expression in CHO 14 nucleotides Optimized constructs for SK-S1Δ peptide expression in CHO 15 nucleotides Optimized constructs for SK-S1-P2 peptide expression in CHO 16 nucleotides Optimized constructs for expression of SK-S1Δ-P2 peptides in CHO 17 nucleotides Optimized constructs for SK-S1-T2 peptide expression in CHO 18 nucleotides Optimized constructs for expression of SK-S1-T2-P2 peptides in CHO 19 peptide human albumin signal peptide 20 peptide Tetanus Toxoid Epitope P2 Domain 21 peptide N protein 22 nucleotides Optimized constructs for N protein expression in BEVS 23 nucleotides Optimized constructs for N protein expression in CHO 24 nucleotides Spike protein full length 25 nucleotides S1 domain 26 nucleotides S2 domain 27 nucleotides RBD domain 28 peptide Spike full length 29 peptide S1 domain 30 peptide S2 domain 31 peptide RBD domain

The (CHO) denotes a nucleic acid sequence optimized for the CHO expression system, (BEVS) denotes a nucleic acid sequence optimized for the BEVS expression system, and is denoted by _CHO and _BEVS in the sequence listing, respectively.

<110> SK bioscience Co., Ltd. <120> Vaccine composition for preventing or treating infection of SARS-CoV-2 comprising a modified Spike protein of SARS-CoV-2 <130> P20-180 <160> 31 <170> KoPatentIn 3.0 <210> 1 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> SK-S1 <400> 1 Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr 1 5 10 15 Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser 20 25 30 Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn 35 40 45 Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys 50 55 60 Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala 65 70 75 80 Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr 85 90 95 Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn 100 105 110 Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu 115 120 125 Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe 130 135 140 Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln 145 150 155 160 Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu 165 170 175 Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser 180 185 190 Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser 195 200 205 Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg 210 215 220 Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp 225 230 235 240 Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr 245 250 255 Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile 260 265 270 Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys 2 75 280 285 Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn 290 295 300 Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr 305 310 315 320 Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser 325 330 335 Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr 340 345 350 Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly 355 360 365 Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala 370 375 380 Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly 385 390 395 400 Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe 405 410 415 Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp S er Lys Val 420 425 430 Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu 435 440 445 Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser 450 455 460 Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln 465 470 475 480 Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg 485 490 495 Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys 500 505 510 Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe 515 520 525 Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys 530 535 540 Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr 545 550 555 560 Asp Ala Val Arg Asp Pro Gln Thr Leu Glu I le Leu Asp Ile Thr Pro 565 570 575 Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser 580 585 590 Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro 595 600 605 Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser 610 615 620 Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala 625 630 635 640 Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly 645 650 655 Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg 660 665 670 Ser <210> 2 <211> 669 <212> PRT <213> Artificial Sequence <220> <223> 4 amino acids are removed from the peptide of SK-S1. <400> 2 Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr 1 5 10 15 Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser 20 25 30 Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn 35 40 45 Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys 50 55 60 Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala 65 70 75 80 Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr 85 90 95 Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn 100 105 110 Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu 115 120 125 Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe 130 135 140 Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln 145 150 155 160 Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu 165 170 175 Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser 180 185 190 Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser 195 200 205 Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg 210 215 220 Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp 225 230 235 240 Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr 245 250 255 Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile 260 265 270 Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys 275 280 285 Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn 290 295 300 Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr 305 310 315 320 Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser 325 330 335 Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr 340 345 350 Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly 355 360 365 Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala 370 375 380 Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly 385 390 395 400 Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe 405 410 415 Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val 420 425 430 Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu 435 440 445 Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser 450 455 460 Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln 465 470 475 480 Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg 485 490 495 Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys 500 505 510 Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe 515 520 525 Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys 530 535 540 Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr 545 550 555 560 Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro 565 570 575 Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser 580 585 590 Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro 595 600 605 Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser 610 615 620 Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala 625 630 635 640 Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly 645 650 655 Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg 660 665 <210> 3 <211> 689 <212> PRT <213> Artificial Sequence <220> <223> SK- S1-P2 <400> 3 Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr 1 5 10 15 Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser 20 25 30 Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn 35 40 45 Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys 50 55 60 Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala 65 70 75 80 Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr 85 90 95 L eu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn 100 105 110 Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu 115 120 125 Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe 130 135 140 Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln 145 150 155 160 Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu 165 170 175 Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser 180 185 190 Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser 195 200 205 Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg 210 215 220 Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp 225 230 235 240 Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr 245 250 255 Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile 260 265 270 Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys 275 280 285 Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn 290 295 300 Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr 305 310 315 320 Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser 325 330 335 Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr 340 345 350 Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly 355 360 365 Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala 370 375 3 80 Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly 385 390 395 400 Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe 405 410 415 Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val 420 425 430 Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu 435 440 445 Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser 450 455 460 Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln 465 470 475 480 Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg 485 490 495 Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys 500 505 510 Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe 515 520 525 Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys 530 535 540 Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr 545 550 555 560 Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro 565 570 575 Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser 580 585 590 Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro 595 600 605 Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser 610 615 620 Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala 625 630 635 640 Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly 645 650 655 Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Gly Ser Gly 660 665 670 Ser Gly Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile Gly Ile Thr Glu 675 680 685 Leu <210> 4 <211> 689 <212> PRT <213> Artificial Sequence <220> <223> P2 is linked to the protein having sequence No.2. <400> 4 Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr 1 5 10 15 Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser 20 25 30 Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn 35 40 45 Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys 50 55 60 Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala 65 70 75 80 Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr 85 90 95 Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn 100 105 110 Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu 115 120 125 Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe 130 135 140 Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln 145 150 155 160 Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu 165 170 175 Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser 180 185 190 Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser 195 200 205 Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg 210 215 220 Phe Gln Thr Leu Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp 225 230 235 240 Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr 245 250 255 Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile 260 265 270 Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys 275 280 285 Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn 290 295 300 Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr 305 310 315 320 Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser 325 330 335 Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr 340 345 350 Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly 355 360 365 Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala 370 375 380 Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly 385 390 395 400 Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe 405 410 415 Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val 420 425 430 Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu 435 440 445 Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser 450 455 460 Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln 465 470 475 480 Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg 485 490 495 Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys 500 505 510 Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe 515 520 525 Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys 530 535 540 Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr 545 550 555 560 Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro 565 570 575 Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser 580 585 590 Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro 595 600 605 Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser 610 615 620 Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala 625 630 635 640 Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly 645 650 655 Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Gly Ser Gly 660 665 670 Ser Gly Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile Gly Ile Thr Glu 675 680 685 Leu <210> 5 <211> 412 <212> PRT <213> Artificial Sequence <220> < 223> SK-S1-T2 <400> 5 Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser 1 5 10 15 Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln 20 25 30 Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro 35 40 45 Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp 50 55 60 Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr 65 70 75 80 Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr 85 90 95 L ys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val 100 105 110 Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys 115 120 125 Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val 130 135 140 Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr 145 150 155 160 Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu 165 170 175 Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn 180 185 190 Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe 195 200 205 Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu 210 215 220 Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys 225 230 235 240 Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly 245 250 255 Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro 260 265 270 Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg 275 280 285 Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly 290 295 300 Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala 305 310 315 320 Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile His 325 330 335 Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn 340 345 350 Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val Asn 355 360 365 Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser 370 375 3 80 Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala Ser 385 390 395 400 Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala 405 410 <210> 6 <211> 432 <212> PRT <213> Artificial Sequence <220> <223> SK-S1-T2-P2 <400> 6 Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser 1 5 10 15 Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln 20 25 30 Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro 35 40 45 Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp 50 55 60 Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr 65 70 75 80 Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr 85 90 95 Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val 100 105 110 Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys 115 120 125 Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val 130 135 140 Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr 145 150 155 160 Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu 165 170 175 Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn 180 185 190 Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe 195 200 205 Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu 210 215 220 Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys 225 230 235 240 Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly 245 250 255 Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro 260 265 270 Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg 275 280 285 Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly 290 295 300 Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala 305 310 315 320 Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile His 325 330 335 Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn 340 345 350 Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val Asn 355 360 365 Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser 370 375 380 Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala Ser 385 390 395 400 Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Gly Ser Gly Ser 405 410 415 Gly Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile Gly Ile Thr Glu Leu 420 425 430 <210> 7 <211> 2076 <212> DNA <213> Artificial Sequence <220> <223> SK-S1_BEVS <400> 7 atgaagtggg tgactttcat ctccctgctg ttcctgttct ccagcgctta cagccagtgc 60 gtcaacctca ccactaggac ccagctgcct cccgcctaca ccaactcttt cactaggggc 120 gtgtactacc ccgacaaggt gttccgctcc tccgtgctgc actctactca ggacctgttc 180 ctgccattct tctcaaacgt gacctggttc cacgctatcc acgtctcagg caccaacgga 240 actaagcgct tcgacaaccc agtgctgcct ttcaacgacg gagtctactt cgcctccacc 300 gaaaagagca acatcatccg tggttggatc ttcggcacca ctctggactc taagactcag 360 tcactgctga tcgtgaacaa cgctaccaac gtggtcatca aggtctgcga gttccagttc 420 tgcaacgacc ccttcctggg tgtgtactac cacaagaaca acaagtcctg gatggagagc 480 gagttccgtg tgtactccag cgctaacaac tgcactttcg agtacgtctc tcagccattc 540 ctgatggacc tggaaggcaa gcagggcaac ttcaagaacc tgagggagtt cgtgttcaag 600 aacatcgacg gctacttcaa gatttactcc aagcacaccc ccatcaacct ggtgcgcgac 660 ctgccacagg gtttcagcgc tctggaacct ctggtggacc tgccaatc gg catcaacatc 720 accaggttcc agactctgct ggctctgcac agaagctacc tgactcctgg agactcttca 780 tccggatgga ccgccggtgc tgccgcttac tacgtgggtt acctgcaacc caggaccttc 840 ctgctgaagt acaacgagaa cggcaccatc actgacgccg tggactgcgc tctggaccca 900 ctgtctgaaa ccaagtgcac tctgaagtca ttcaccgtcg agaagggcat ctaccagact 960 tctaacttcc gcgtgcagcc taccgaatca atcgtccgtt tcccaaacat cactaacctg 1020 tgccctttcg gagaggtgtt caacgccacc agattcgctt ccgtctacgc ctggaaccgc 1080 aagcgtatct ctaactgcgt cgctgactac tcagtgctgt acaacagcgc ctctttctca 1140 accttcaagt gctacggagt gtctcctact aagctgaacg acctgtgctt caccaacgtc 1200 tacgctgact cattcgtgat ccgcggtgac gaggtccgtc agatcgctcc cggacagact 1260 ggcaagatcg ccgactacaa ctacaagctg ccagacgact tcaccggttg cgtgatcgcc 1320 tggaactcta acaacctgga ctcaaaggtc ggtggcaact acaactacct gtacaggctg 1380 ttcagaaagt ctaacctgaa gcctttcgag cgcgacatct ccactgaaat ctaccaggct 1440 ggtagcaccc cctgcaacgg cgtggaagga ttcaactgct acttccctct gcaatcatac 1500 ggcttccagc ccactaacgg cgtcggatac cagccatacc gtgtggtcgt gctgtcct tc 1560 gagctgctcc acgctcctgc tactgtgtgc ggccccaaga agagcaccaa cctggtcaag 1620 aacaagtgcg tgaacttcaa cttcaacgga ctgaccggta ctggcgtgct gaccgaatcc 1680 aacaagaagt tcctgccttt ccagcagttc ggtcgcgaca tcgctgacac cactgacgcc 1740 gtccgtgacc ctcagaccct ggagatcctg gacatcactc cctgctcctt cggaggtgtc 1800 agcgtgatca ctcccggcac caacactagc aaccaggtcg ctgtgctgta ccaggacgtg 1860 aactgcaccg aagtcccagt ggccatccac gctgaccagc tgacccctac ttggagggtg 1920 tactctaccg gctcaaacgt gttccagact agagctggct gcctgatcgg agccgagcac 1980 gtcaacaact cctacgaatg cgacatccca atcggagctg gtatctgcgc ctcctaccag 2040 acccagacta acagccctag gcgtgctagg tcctaa 2076 <210> 8 <211> 2064 <212> DNA <213> Artificial Sequence <220> <223> Codons for expression of the protein having sequence No.2. in BEVS <400> 8 atgaagtggg tgactttcat ctccctgctg ttcctgttct ccagcgctta cagccagtgc 60 gtcaacctca ccactaggac ccagctgcct cccgcctaca ccaactcttt cactaggggc 120 gtgtactacc ccgacaaggt gttccgctcc tccgtgctgc actctactca ggacctgttc 180 ctgccattct tctcaaacgt gacctggttc cacgctatcc acgtctcagg caccaacgga 240 actaagcgct tcgacaaccc agtgctgcct ttcaacgacg gagtctactt cgcctccacc 300 gaaaagagca acatcatccg tggttggatc ttcggcacca ctctggactc taagactcag 360 tcactgctga tcgtgaacaa cgctaccaac gtggtcatca aggtctgcga gttccagttc 420 tgcaacgacc ccttcctggg tgtgtactac cacaagaaca acaagtcctg gatggagagc 480 gagttccgtg tgtactccag cgctaacaac tgcactttcg agtacgtctc tcagccattc 540 ctgatggacc tggaaggcaa gcagggcaac ttcaagaacc tgagggagtt cgtgttcaag 600 aacatcgacg gctacttcaa gatttactcc aagcacaccc ccatcaacct ggtgcgcgac 660 ctgccacagg gtttcagcgc tctggaacct ctggtggacc tgccaatcgg catcaacatc 720 accaggttcc agactctgct ggctctgcac agaagctacc tgactcctgg agactcttca 780 tccggatgga ccgccggtgc tgccgcttac tacgtgggtt acctgcaacc caggaccttc 840 ctgct gaagt acaacgagaa cggcaccatc actgacgccg tggactgcgc tctggaccca 900 ctgtctgaaa ccaagtgcac tctgaagtca ttcaccgtcg agaagggcat ctaccagact 960 tctaacttcc gcgtgcagcc taccgaatca atcgtccgtt tcccaaacat cactaacctg 1020 tgccctttcg gagaggtgtt caacgccacc agattcgctt ccgtctacgc ctggaaccgc 1080 aagcgtatct ctaactgcgt cgctgactac tcagtgctgt acaacagcgc ctctttctca 1140 accttcaagt gctacggagt gtctcctact aagctgaacg acctgtgctt caccaacgtc 1200 tacgctgact cattcgtgat ccgcggtgac gaggtccgtc agatcgctcc cggacagact 1260 ggcaagatcg ccgactacaa ctacaagctg ccagacgact tcaccggttg cgtgatcgcc 1320 tggaactcta acaacctgga ctcaaaggtc ggtggcaact acaactacct gtacaggctg 1380 ttcagaaagt ctaacctgaa gcctttcgag cgcgacatct ccactgaaat ctaccaggct 1440 ggtagcaccc cctgcaacgg cgtggaagga ttcaactgct acttccctct gcaatcatac 1500 ggcttccagc ccactaacgg cgtcggatac cagccatacc gtgtggtcgt gctgtccttc 1560 gagctgctcc acgctcctgc tactgtgtgc ggccccaaga agagcaccaa cctggtcaag 1620 aacaagtgcg tgaacttcaa cttcaacgga ctgaccggta ctggcgtgct gaccgaatcc 1680 aacaagaagt tc ctgccttt ccagcagttc ggtcgcgaca tcgctgacac cactgacgcc 1740 gtccgtgacc ctcagaccct ggagatcctg gacatcactc cctgctcctt cggaggtgtc 1800 agcgtgatca ctcccggcac caacactagc aaccaggtcg ctgtgctgta ccaggacgtg 1860 aactgcaccg aagtcccagt ggccatccac gctgaccagc tgacccctac ttggagggtg 1920 tactctaccg gctcaaacgt gttccagact agagctggct gcctgatcgg agccgagcac 1980 gtcaacaact cctacgaatg cgacatccca atcggagctg gtatctgcgc ctcctaccag 2040 acccagacta acagccctag gtaa 2064 <210> 9 < 211> 2136 <212> DNA <213> Artificial Sequence <220> <223> SK-S1-P2_BEVS <400> 9 atgaagtggg tgactttcat ctccctgctg ttcctgttct ccagcgctta cagccagtgc 60 gtcaacctca ccactaggac ccagctgcct cccgcctaca ccaactcttt cactaggggc 120 gtgtactacc ccgacaaggt gttccgctcc tccgtgctgc actctactca ggacctgttc 180 ctgccattct tctcaaacgt gacctggttc cacgctatcc acgtctcagg caccaacgga 240 actaagcgct tcgacaaccc agtgctgcct ttcaacgacg gagtctactt cgcctccacc 300 gaaaagagca acattcatccg tggacttggatc cat aggact cactggacttggatc ttcggtggatc cacta acctggtggatc ttcggcacca c tctgcga gttccagttc 420 tgcaacgacc ccttcctggg tgtgtactac cacaagaaca acaagtcctg gatggagagc 480 gagttccgtg tgtactccag cgctaacaac tgcactttcg agtacgtctc tcagccattc 540 ctgatggacc tggaaggcaa gcagggcaac ttcaagaacc tgagggagtt cgtgttcaag 600 aacatcgacg gctacttcaa gatttactcc aagcacaccc ccatcaacct ggtgcgcgac 660 ctgccacagg gtttcagcgc tctggaacct ctggtggacc tgccaatcgg catcaacatc 720 accaggttcc agactctgct ggctctgcac agaagctacc tgactcctgg agactcttca 780 tccggatgga ccgccggtgc tgccgcttac tacgtgggtt acctgcaacc caggaccttc 840 ctgctgaagt acaacgagaa cggcaccatc actgacgccg tggactgcgc tctggaccca 900 ctgtctgaaa ccaagtgcac tctgaagtca ttcaccgtcg agaagggcat ctaccagact 960 tctaacttcc gcgtgcagcc taccgaatca atcgtccgtt tcccaaacat cactaacctg 1020 tgccctttcg gagaggtgtt caacgccacc agattcgctt ccgtctacgc ctggaaccgc 1080 aagcgtatct ctaactgcgt cgctgactac tcagtgctgt acaacagcgc ctctttctca 1140 accttcaagt gctacggagt gtctcctact aagctgaacg acctgtgctt caccaacgtc 1200 tacgctgact cattcgtgat ccgcggtgac gaggtccgtc agatcgctcc cggacaga ct 1260 ggcaagatcg ccgactacaa ctacaagctg ccagacgact tcaccggttg cgtgatcgcc 1320 tggaactcta acaacctgga ctcaaaggtc ggtggcaact acaactacct gtacaggctg 1380 ttcagaaagt ctaacctgaa gcctttcgag cgcgacatct ccactgaaat ctaccaggct 1440 ggtagcaccc cctgcaacgg cgtggaagga ttcaactgct acttccctct gcaatcatac 1500 ggcttccagc ccactaacgg cgtcggatac cagccatacc gtgtggtcgt gctgtccttc 1560 gagctgctcc acgctcctgc tactgtgtgc ggccccaaga agagcaccaa cctggtcaag 1620 aacaagtgcg tgaacttcaa cttcaacgga ctgaccggta ctggcgtgct gaccgaatcc 1680 aacaagaagt tcctgccttt ccagcagttc ggtcgcgaca tcgctgacac cactgacgcc 1740 gtccgtgacc ctcagaccct ggagatcctg gacatcactc cctgctcctt cggaggtgtc 1800 agcgtgatca ctcccggcac caacactagc aaccaggtcg ctgtgctgta ccaggacgtg 1860 aactgcaccg aagtcccagt ggccatccac gctgaccagc tgacccctac ttggagggtg 1920 tactctaccg gctcaaacgt gttccagact agagctggct gcctgatcgg agccgagcac 1980 gtcaacaact cctacgaatg cgacatccca atcggagctg gtatctgcgc ctcctaccag 2040 acccagacta acagccctag gcgtgctagg tccggctccg gaagcggaca gtacatcaag 210 0 gccaacagca agttcatcgg tatcaccgag ctgtaa 2136 <210> 10 <211> 2124 <212> DNA <213> Artificial Sequence <220> <223> Codons for expression of the protein having sequence No.4. in BEVS <400> 10 atgaagtggg tgactttcat ctccctgctg ttcctgttct ccagcgctta cagccagtgc 60 gtcaacctca ccactaggac ccagctgcct cccgcctaca ccaactcttt cactaggggc 120 gtgtactacc ccgacaaggt gttccgctcc tccgtgctgc actctactca ggacctgttc 180 ctgccattct tctcaaacgt gacctggttc cacgctatcc acgtctcagg caccaacgga 240 actaagcgct tcgacaaccc agtgctgcct ttcaacgacg gagtctactt cgcctccacc 300 gaaaagagca acatcatccg tggttggatc ttcggcacca ctctggactc taagactcag 360 tcactgctga tcgtgaacaa cgctaccaac gtggtcatca aggtctgcga gttccagttc 420 tgcaacgacc ccttcctggg tgtgtactac cacaagaaca acaagtcctg gatggagagc 480 gagttccgtg tgtactccag cgctaacaac tgcactttcg agtacgtctc tcagccattc 540 ctgatggacc tggaaggcaa gcagggcaac ttcaagaacc tgagggagtt cgtgttcaag 600 aacatcgacg gctacttcaa gatttactcc aagcacaccc ccatcaacct ggtgcgcgac 660 ctgccacagg gtttcagcgc tctggaacct ctggtggacc tgccaatcgg catcaacatc 720 accaggttcc agactctgct ggctctgcac agaagctacc tgactcctgg agactcttca 780 tccggatgga ccgccggtgc tgccgcttac tacgtgggtt acctgcaacc caggaccttc 840 ctgctgaagt acaacgagaa cggcaccatc actgacgccg tggactgcgc tctggaccca 900 ctgtctgaaa ccaagtgcac tctgaagtca ttcaccgtcg agaagggcat ctaccagact 960 tctaacttcc gcgtgcagcc taccgaatca atcgtccgtt tcccaaacat cactaacctg 1020 tgccctttcg gagaggtgtt caacgccacc agattcgctt ccgtctacgc ctggaaccgc 1080 aagcgtatct ctaactgcgt cgctgactac tcagtgctgt acaacagcgc ctctttctca 1140 accttcaagt gctacg gagt gtctcctact aagctgaacg acctgtgctt caccaacgtc 1200 tacgctgact cattcgtgat ccgcggtgac gaggtccgtc agatcgctcc cggacagact 1260 ggcaagatcg ccgactacaa ctacaagctg ccagacgact tcaccggttg cgtgatcgcc 1320 tggaactcta acaacctgga ctcaaaggtc ggtggcaact acaactacct gtacaggctg 1380 ttcagaaagt ctaacctgaa gcctttcgag cgcgacatct ccactgaaat ctaccaggct 1440 ggtagcaccc cctgcaacgg cgtggaagga ttcaactgct acttccctct gcaatcatac 1500 ggcttccagc ccactaacgg cgtcggatac cagccatacc gtgtggtcgt gctgtccttc 1560 gagctgctcc acgctcctgc tactgtgtgc ggccccaaga agagcaccaa cctggtcaag 1620 aacaagtgcg tgaacttcaa cttcaacgga ctgaccggta ctggcgtgct gaccgaatcc 1680 aacaagaagt tcctgccttt ccagcagttc ggtcgcgaca tcgctgacac cactgacgcc 1740 gtccgtgacc ctcagaccct ggagatcctg gacatcactc cctgctcctt cggaggtgtc 1800 agcgtgatca ctcccggcac caacactagc aaccaggtcg ctgtgctgta ccaggacgtg 1860 aactgcaccg aagtcccagt ggccatccac gctgaccagc tgacccctac ttggagggtg 1920 tactctaccg gctcaaacgt gttccagact agagctggct gcctgatcgg agccgagcac 1980 gtcaacaact cctacgaatg c gacatccca atcggagctg gtatctgcgc ctcctaccag 2040 acccagacta acagccctag gggctccgga agcggacagt acatcaaggc caacagcaag 2100 ttcatcggta tcaccgagct gtaa 2124 <210> 11 <211> SKTg < 1293 <212> 11 atgaStact Artificial Sequence ctccctgctg ttcctgttct ccagcgctta cagcgactgc 60 gctctggacc cactgtctga aaccaagtgc actctgaagt cattcaccgt cgagaagggc 120 atctaccaga cttctaactt ccgcgtgcag cctaccgaat caatcgtccg tttcccaaac 180 atcactaacc tgtgcccttt cggagaggtg ttcaacgcca ccagattcgc ttccgtctac 240 gcctggaacc gcaagcgtat ctctaactgc gtcgctgact actcagtgct gtacaacagc 300 gcctctttct caaccttcaa gtgctacgga gtgtctccta ctaagctgaa cgacctgtgc 360 ttcaccaacg tctacgctga ctcattcgtg atccgcggtg acgaggtccg tcagatcgct 420 cccggacaga ctggcaagat cgccgactac aactacaagc tgccagacga cttcaccggt 480 tgcgtgatcg cctggaactc taacaacctg gactcaaagg tcggtggcaa ctacaactac 540 ctgtacaggc tgttcagagaa gtctaacctg aagcctgcgtgatg acctgatt g acctaacctg aagcctcacttcg acctg ttccct 660 ctgcaatcat acggcttcca gcccactaac ggcgtcggat accagccata ccgtgtggtc 720 gtgctgtcct tcgagctgct ccacgctcct gctactgtgt gcggccccaa gaagagcacc 780 aacctggtca agaacaagtg cgtgaacttc aacttcaacg gactgaccgg tactggcgtg 840 ctgaccgaat ccaacaagaa gttcctgcct ttccagcagt tcggtcgcga catcgctgac 900 accactgacg ccgtccgtga ccctcagacc ctggagatcc tggacatcac tccctgctcc 960 ttcggaggtg tcagcgtgat cactcccggc accaacacta gcaaccaggt cgctgtgctg 1020 taccaggacg tgaactgcac cgaagtccca gtggccatcc acgctgacca gctgacccct 1080 acttggaggg tgtactctac cggctcaaac gtgttccaga ctagagctgg ctgcctgatc 1140 ggagccgagc acgtcaacaa ctcctacgaa tgcgacatcc caatcggagc tggtatctgc 1200 gcctcctacc agacccagac taacagccct aggcgggcac gtagtgtagc tagtcaatcc 1260 atcattgcct acactatgtc acttggtgca taa 1293 <210> 12 <211> 1353 <212> DNA <213> Artificial Sequence <220> <223> SK-S1-T2-P2_BEVS <400> 12 atgaagtggg tgactttcat ctccctgctg ttcctgttct ccagcgctta cagcgactgc 60 gctctggacc cactgtctga aaccaagtgc actctgaagt cattcaccgt cgagaagggc 120 atctaccag a cttctaactt ccgcgtgcag cctaccgaat caatcgtccg tttcccaaac 180 atcactaacc tgtgcccttt cggagaggtg ttcaacgcca ccagattcgc ttccgtctac 240 gcctggaacc gcaagcgtat ctctaactgc gtcgctgact actcagtgct gtacaacagc 300 gcctctttct caaccttcaa gtgctacgga gtgtctccta ctaagctgaa cgacctgtgc 360 ttcaccaacg tctacgctga ctcattcgtg atccgcggtg acgaggtccg tcagatcgct 420 cccggacaga ctggcaagat cgccgactac aactacaagc tgccagacga cttcaccggt 480 tgcgtgatcg cctggaactc taacaacctg gactcaaagg tcggtggcaa ctacaactac 540 ctgtacaggc tgttcagaaa gtctaacctg aagcctttcg agcgcgacat ctccactgaa 600 atctaccagg ctggtagcac cccctgcaac ggcgtggaag gattcaactg ctacttccct 660 ctgcaatcat acggcttcca gcccactaac ggcgtcggat accagccata ccgtgtggtc 720 gtgctgtcct tcgagctgct ccacgctcct gctactgtgt gcggccccaa gaagagcacc 780 aacctggtca agaacaagtg cgtgaacttc aacttcaacg gactgaccgg tactggcgtg 840 ctgaccgaat ccaacaagaa gttcctgcct ttccagcagt tcggtcgcga catcgctgac 900 accactgacg ccgtccgtga ccctcagacc ctggagatcc tggacatcac tccctgctcc 960 ttcggaggtg tcagcgtgat cactccc ggc accaacacta gcaaccaggt cgctgtgctg 1020 taccaggacg tgaactgcac cgaagtccca gtggccatcc acgctgacca gctgacccct 1080 acttggaggg tgtactctac cggctcaaac gtgttccaga ctagagctgg ctgcctgatc 1140 ggagccgagc acgtcaacaa ctcctacgaa tgcgacatcc caatcggagc tggtatctgc 1200 gcctcctacc agacccagac taacagccct aggcgggcac gtagtgtagc tagtcaatcc 1260 atcattgcct acactatgtc acttggtgca ggctccggaa gcggacagta catcaaggcc 1320 aacagcaagt tcatcggtat caccgagctg taa 1353 <210> 13 < 211> 2061 <212> DNA <213> Artificial Sequence <220> <223> SK-S1_CHO <400> 13 atgttcgtgt ttctggtgct gctgccactg gtgtccagcc agtgcgtgaa tctgaccaca 60 agaacccagc tgccccctgc ctataccaac agctttacaa ggggcgtgta ctatcctgat 120 aaggtgttca ggtcctccgt gctgcacagc acacaggacc tgtttctgcc attcttttct 180 aatgtgacct ggttccacgc catccacgtg tccggcacca acggcacaaa gagatttgat 240 aatccagtgc tgccctttaa cgacggcgtg tacttcgctt ctaccgagaa gtccaacatc 300 atccgcggct ggatcttcgg caccacactg gatagcaaga cacagtctct gctgatcgtg gctgatcgtg aacaactggt cgaccctttt 420 ctgggcgtgt actatcataa gaacaataag tcctggatgg agagcgagtt ccgcgtgtat 480 agctctgcta acaattgtac atttgagtac gtgtctcagc cattcctgat ggatctggag 540 ggcaagcagg gcaacttcaa gaacctgagg gagttcgtgt ttaagaatat cgacggctac 600 ttcaaaatct actctaagca cacccctatc aacctggtgc gggatctgcc acagggcttt 660 tccgctctgg agcctctggt ggacctgcca atcggcatca acatcaccag gttccagaca 720 ctgctggccc tgcatcggtc ctacctgaca cccggcgatt ccagctctgg atggaccgct 780 ggcgccgctg cctactatgt gggctacctc cagcctagga cctttctgct gaagtacaac 840 gagaatggca ccatcacaga cgctgtggat tgcgccctgg acccactgag cgagacaaag 900 tgtacactga agtctttcac cgtggagaag ggcatctatc agacatctaa cttcagggtg 960 cagcccaccg agtccatcgt gcgcttccca aatatcacaa acctgtgccc ctttggcgag 1020 gtgttcaacg ctacccggtt tgcctccgtg tacgcttgga atagaaagcg catctctaac 1080 tgcgtggccg attattccgt gctgtacaac tccgcctcct tctccacctt caagtgctat 1140 ggcgtgagcc ccacaaagct gaatgacctg tgctttacca acgtgtacgc cgattctttc 1200 gtgatcagag gcgacgaggt gcgccagatc gcccctggcc agacaggcaa gatcgctgat 1260tacaattata agctgccaga cgatttcacc ggctgcgtga tcgcttggaa ctctaacaat 1320 ctggactcca aagtgggcgg caactacaat tatctgtaca ggctgtttcg gaagtctaat 1380 ctgaagccct tcgagagaga catctccaca gaaatctacc aggccggcag caccccttgc 1440 aatggcgtgg agggctttaa ctgttatttc cccctccagt cctacggctt tcagcctacc 1500 aacggcgtgg gctatcagcc ataccgcgtg gtggtgctgt ccttcgagct gctgcacgct 1560 ccagctacag tgtgcggccc taagaagagc accaatctgg tgaagaacaa gtgcgtgaac 1620 ttcaacttca acggcctgac cggcacaggc gtgctgaccg agtccaataa gaagttcctg 1680 ccctttcagc agttcggcag agacatcgcc gataccacag acgctgtgcg cgatccccag 1740 accctggaga tcctggacat cacaccttgc agctttggcg gcgtgtctgt gatcacacct 1800 ggcaccaata caagcaacca ggtggccgtg ctgtatcagg atgtgaattg taccgaggtg 1860 ccagtggcta tccacgccga ccagctgacc cccacatgga gggtgtactc caccggcagc 1920 aacgtgttcc agacacgggc cggatgtctg atcggagctg agcatgtgaa caattcctat 1980 gagtgcgaca tcccaatcgg cgccggcatc tgtgcctcct accagaccca gacaaactcc 2040 cccaggcggg ctcggagctg a 2061 <210> 14 < 211> 2049 <212> DNA <213> Artificial S sequence <220> <223> Codons for expression of the protein having sequence No.2. in CHO <400> 14 atgttcgtgt ttctggtgct gctgccactg gtgtccagcc agtgcgtgaa tctgaccaca 60 agaacccagc tgccccctgc ctataccaac agctttacaa ggggcgtgta ctatcctgat 120 aaggtgttca ggtcctccgt gctgcacagc acacaggacc tgtttctgcc attcttttct 180 aatgtgacct ggttccacgc catccacgtg tccggcacca acggcacaaa gagatttgat 240 aatccagtgc tgccctttaa cgacggcgtg tacttcgctt ctaccgagaa gtccaacatc 300 atccgcggct ggatcttcgg caccacactg gatagcaaga cacagtctct gctgatcgtg 360 aacaatgcca ccaacgtggt catcaaggtg tgcgagttcc agttttgtaa cgaccctttt 420 ctgggcgtgt actatcataa gaacaataag tcctggatgg agagcgagtt ccgcgtgtat 480 agctctgcta acaattgtac atttgagtac gtgtctcagc cattcctgat ggatctggag 540 ggcaagcagg gcaacttcaa gaacctgagg gagttcgtgt ttaagaatat cgacggctac 600 ttcaaaatct actctaagca cacccctatc aacctggtgc gggatctgcc acagggcttt 660 tccgctctgg agcctctggt ggacctgcca atcggcatca acatcaccag gttccagaca 720 ctgctggccc tgcatcggtc ctacctgaca cccggcgatt ccagctctgg atggaccgct 780 ggcgccgctg cctactatgt gggctacctc cagcctagga cctttctgct gaagtacaac 840 gagaa tggca ccatcacaga cgctgtggat tgcgccctgg acccactgag cgagacaaag 900 tgtacactga agtctttcac cgtggagaag ggcatctatc agacatctaa cttcagggtg 960 cagcccaccg agtccatcgt gcgcttccca aatatcacaa acctgtgccc ctttggcgag 1020 gtgttcaacg ctacccggtt tgcctccgtg tacgcttgga atagaaagcg catctctaac 1080 tgcgtggccg attattccgt gctgtacaac tccgcctcct tctccacctt caagtgctat 1140 ggcgtgagcc ccacaaagct gaatgacctg tgctttacca acgtgtacgc cgattctttc 1200 gtgatcagag gcgacgaggt gcgccagatc gcccctggcc agacaggcaa gatcgctgat 1260 tacaattata agctgccaga cgatttcacc ggctgcgtga tcgcttggaa ctctaacaat 1320 ctggactcca aagtgggcgg caactacaat tatctgtaca ggctgtttcg gaagtctaat 1380 ctgaagccct tcgagagaga catctccaca gaaatctacc aggccggcag caccccttgc 1440 aatggcgtgg agggctttaa ctgttatttc cccctccagt cctacggctt tcagcctacc 1500 aacggcgtgg gctatcagcc ataccgcgtg gtggtgctgt ccttcgagct gctgcacgct 1560 ccagctacag tgtgcggccc taagaagagc accaatctgg tgaagaacaa gtgcgtgaac 1620 ttcaacttca acggcctgac cggcacaggc gtgctgaccg agtccaataa gaagttcctg 1680 ccctttcagc ag ttcggcag agacatcgcc gataccacag acgctgtgcg cgatccccag 1740 accctggaga tcctggacat cacaccttgc agctttggcg gcgtgtctgt gatcacacct 1800 ggcaccaata caagcaacca ggtggccgtg ctgtatcagg atgtgaattg taccgaggtg 1860 ccagtggcta tccacgccga ccagctgacc cccacatgga gggtgtactc caccggcagc 1920 aacgtgttcc agacacgggc cggatgtctg atcggagctg agcatgtgaa caattcctat 1980 gagtgcgaca tcccaatcgg cgccggcatc tgtgcctcct accagaccca gacaaactcc 2040 cccaggtga 2049 <210> 15 <211> 2121 <212> DNA <213> Artificial Sequence <220> <223> SK-S1-P2_CHO <400> 15 atgttcgtgt ttctggtgct gctgccactg gtgtccagcc agtgcgtgaa tctgaccaca 60 agaacccagc tgccccctgc ctataccaac agctttacaa ggggcgtgta ctatcctgat 120 aaggtgttca ggtcctccgt gctgcacagc acacaggacc tgtttctgcc attcttttct 180 aatgtgacct ggttccacgc catccacgtg tccggcacca acggcacaaa gagatttgat 240 aatccagtgc tgccctttaa cgacggcgtg tacttcgctt ctaccgagaa gtccaacatc 300 atccgcggct ggatcttcgg caccacactg gatagcaaga cacagtctct gctgatcgt cc acctggt cacaatgt tt 420 ctgggcgtgt actatcataa gaacaataag tcctggatgg agagcgagtt ccgcgtgtat 480 agctctgcta acaattgtac atttgagtac gtgtctcagc cattcctgat ggatctggag 540 ggcaagcagg gcaacttcaa gaacctgagg gagttcgtgt ttaagaatat cgacggctac 600 ttcaaaatct actctaagca cacccctatc aacctggtgc gggatctgcc acagggcttt 660 tccgctctgg agcctctggt ggacctgcca atcggcatca acatcaccag gttccagaca 720 ctgctggccc tgcatcggtc ctacctgaca cccggcgatt ccagctctgg atggaccgct 780 ggcgccgctg cctactatgt gggctacctc cagcctagga cctttctgct gaagtacaac 840 gagaatggca ccatcacaga cgctgtggat tgcgccctgg acccactgag cgagacaaag 900 tgtacactga agtctttcac cgtggagaag ggcatctatc agacatctaa cttcagggtg 960 cagcccaccg agtccatcgt gcgcttccca aatatcacaa acctgtgccc ctttggcgag 1020 gtgttcaacg ctacccggtt tgcctccgtg tacgcttgga atagaaagcg catctctaac 1080 tgcgtggccg attattccgt gctgtacaac tccgcctcct tctccacctt caagtgctat 1140 ggcgtgagcc ccacaaagct gaatgacctg tgctttacca acgtgtacgc cgattctttc 1200 gtgatcagag gcgacgaggt gcgccagatc gcccctggcc agacaggcaa gatcgctgat 1260 tacaatta ta agctgccaga cgatttcacc ggctgcgtga tcgcttggaa ctctaacaat 1320 ctggactcca aagtgggcgg caactacaat tatctgtaca ggctgtttcg gaagtctaat 1380 ctgaagccct tcgagagaga catctccaca gaaatctacc aggccggcag caccccttgc 1440 aatggcgtgg agggctttaa ctgttatttc cccctccagt cctacggctt tcagcctacc 1500 aacggcgtgg gctatcagcc ataccgcgtg gtggtgctgt ccttcgagct gctgcacgct 1560 ccagctacag tgtgcggccc taagaagagc accaatctgg tgaagaacaa gtgcgtgaac 1620 ttcaacttca acggcctgac cggcacaggc gtgctgaccg agtccaataa gaagttcctg 1680 ccctttcagc agttcggcag agacatcgcc gataccacag acgctgtgcg cgatccccag 1740 accctggaga tcctggacat cacaccttgc agctttggcg gcgtgtctgt gatcacacct 1800 ggcaccaata caagcaacca ggtggccgtg ctgtatcagg atgtgaattg taccgaggtg 1860 ccagtggcta tccacgccga ccagctgacc cccacatgga gggtgtactc caccggcagc 1920 aacgtgttcc agacacgggc cggatgtctg atcggagctg agcatgtgaa caattcctat 1980 gagtgcgaca tcccaatcgg cgccggcatc tgtgcctcct accagaccca gacaaactcc 2040 cccaggcggg ctcggagcgg ctccggctcc ggccagtaca tcaaggccaa ctccaagttc 2100 atcggcatca ccg agctgta a 2121 <210> 16 <211> 2109 <212> DNA <213> Artificial Sequence <220> <223> Codons for expression of the protein having sequence No.4. in CHO. <400> 16 atgttcgtgt ttctggtgct gctgccactg gtgtccagcc agtgcgtgaa tctgaccaca 60 agaacccagc tgccccctgc ctataccaac agctttacaa ggggcgtgta ctatcctgat 120 aaggtgttca ggtcctccgt gctgcacagc acacaggacc tgtttctgcc attcttttct 180 aatgtgacct ggttccacgc catccacgtg tccggcacca acggcacaaa gagatttgat 240 aatccagtgc tgccctttaa cgacggcgtg tacttcgctt ctaccgagaa gtccaacatc 300 atccgcggct ggatcttcgg caccacactg gatagcaaga cacagtctct gctgatcgtg 360 aacaatgcca ccaacgtggt catcaaggtg tgcgagttcc agttttgtaa cgaccctttt 420 ctgggcgtgt actatcataa gaacaataag tcctggatgg agagcgagtt ccgcgtgtat 480 agctctgcta acaattgtac atttgagtac gtgtctcagc cattcctgat ggatctggag 540 ggcaagcagg gcaacttcaa gaacctgagg gagttcgtgt ttaagaatat cgacggctac 600 ttcaaaatct actctaagca cacccctatc aacctggtgc gggatctgcc acagggcttt 660 tccgctctgg agcctctggt ggacctgcca atcggcatca acatcaccag gttccagaca 720 ctgctggccc tgcatcggtc ctacctgaca cccggcgatt ccagctctgg atggaccgct 780 ggcgccgctg cctactatgt gggctacctc cagcctagga cctttctgct gaagtacaac 840 gagaatggca c catcacaga cgctgtggat tgcgccctgg acccactgag cgagacaaag 900 tgtacactga agtctttcac cgtggagaag ggcatctatc agacatctaa cttcagggtg 960 cagcccaccg agtccatcgt gcgcttccca aatatcacaa acctgtgccc ctttggcgag 1020 gtgttcaacg ctacccggtt tgcctccgtg tacgcttgga atagaaagcg catctctaac 1080 tgcgtggccg attattccgt gctgtacaac tccgcctcct tctccacctt caagtgctat 1140 ggcgtgagcc ccacaaagct gaatgacctg tgctttacca acgtgtacgc cgattctttc 1200 gtgatcagag gcgacgaggt gcgccagatc gcccctggcc agacaggcaa gatcgctgat 1260 tacaattata agctgccaga cgatttcacc ggctgcgtga tcgcttggaa ctctaacaat 1320 ctggactcca aagtgggcgg caactacaat tatctgtaca ggctgtttcg gaagtctaat 1380 ctgaagccct tcgagagaga catctccaca gaaatctacc aggccggcag caccccttgc 1440 aatggcgtgg agggctttaa ctgttatttc cccctccagt cctacggctt tcagcctacc 1500 aacggcgtgg gctatcagcc ataccgcgtg gtggtgctgt ccttcgagct gctgcacgct 1560 ccagctacag tgtgcggccc taagaagagc accaatctgg tgaagaacaa gtgcgtgaac 1620 ttcaacttca acggcctgac cggcacaggc gtgctgaccg agtccaataa gaagttcctg 1680 ccctttcagc agttcggca g agacatcgcc gataccacag acgctgtgcg cgatccccag 1740 accctggaga tcctggacat cacaccttgc agctttggcg gcgtgtctgt gatcacacct 1800 ggcaccaata caagcaacca ggtggccgtg ctgtatcagg atgtgaattg taccgaggtg 1860 ccagtggcta tccacgccga ccagctgacc cccacatgga gggtgtactc caccggcagc 1920 aacgtgttcc agacacgggc cggatgtctg atcggagctg agcatgtgaa caattcctat 1980 gagtgcgaca tcccaatcgg cgccggcatc tgtgcctcct accagaccca gacaaactcc 2040 cccaggggct ccggctccgg ccagtacatc aaggccaact ccaagttcat cggcatcacc 2100 gagctgtaa 2109 <210> 17 <211> 1293 <212> DNA <213> Artificial Sequence <220> <223> SK-S1-T2_CHO <400> 17 atgaagtggg tcactttcat cagcctgttg tttctgttca gctccgccta ctctgattgctta 60 gccctggacc cactgagcgt cacc accctggacc cactgagcga ccatcgtgcg cttcccaaat 180 atcacaaacc tgtgcccctt tggcgaggtg ttcaacgcta cccggtttgc ctccgtgtac 240 gcttggaata gaaagcgcat ctctaactgc cc gtggccgatt attccgtatgct gtacaactg cc cc gtggccgatt attccgtatgct gtacaactg cc tttaccaacg tgtacgccga ttctttcgtg atcagaggcg acgaggtgcg ccagatcgcc 420 cctggccaga caggcaagat cgctgattac aattataagc tgccagacga tttcaccggc 480 tgcgtgatcg cttggaactc taacaatctg gactccaaag tgggcggcaa ctacaattat 540 ctgtacaggc tgtttcggaa gtctaatctg aagcccttcg agagagacat ctccacagaa 600 atctaccagg ccggcagcac cccttgcaat ggcgtggagg gctttaactg ttatttcccc 660 ctccagtcct acggctttca gcctaccaac ggcgtgggct atcagccata ccgcgtggtg 720 gtgctgtcct tcgagctgct gcacgctcca gctacagtgt gcggccctaa gaagagcacc 780 aatctggtga agaacaagtg cgtgaacttc aacttcaacg gcctgaccgg cacaggcgtg 840 ctgaccgagt ccaataagaa gttcctgccc tttcagcagt tcggcagaga catcgccgat 900 accacagacg ctgtgcgcga tccccagacc ctggagatcc tggacatcac accttgcagc 960 tttggcggcg tgtctgtgat cacacctggc accaatacaa gcaaccaggt ggccgtgctg 1020 tatcaggatg tgaattgtac cgaggtgcca gtggctatcc acgccgacca gctgaccccc 1080 acatggaggg tgtactccac cggcagcaac gtgttccaga cacgggccgg atgtctgatc 1140 ggagctgagc atgtgaacaa ttcctatgag tgcgacatcc caatcggcgc cggcatctgt 1200 gcctcctacc agacccagac aaacagccca aggcgggcca ggtctgtggc ttcccagagc 1260 atcatcgcct atac catgtc cctgggcgcc taa 1293 <210> 18 <211> 1353 <212> DNA <213> Artificial Sequence <220> <223> SK-S1-T2-P2_CHO <400> 18 atgaagtggg tcactttcat cagcctgttg ttttctctgtttca gctccgccta gccctagtacctgtgc acctgactagtgtgtcc acctgactagtgtgtgc 60 gccctagtacctgccta ggagaagggc 120 atctatcaga catctaactt cagggtgcag cccaccgagt ccatcgtgcg cttcccaaat 180 atcacaaacc tgtgcccctt tggcgaggtg ttcaacgcta cccggtttgc ctccgtgtac 240 gcttggaata gaaagcgcat ctctaactgc gtggccgatt attccgtgct gtacaactcc 300 gcctccttct ccaccttcaa gtgctatggc gtgagcccca caaagctgaa tgacctgtgc 360 tttaccaacg tgtacgccga ttctttcgtg atcagaggcg acgaggtgcg ccagatcgcc 420 cctggccaga caggcaagat cgctgattac aattataagc tgccagacga tttcaccggc 480 tgcgtgatcg cttggaactc taacaatctg gactccaaag tgggcggcaa ctacaattat 540 ctgtacaggc tgtttcggaa gtctaatctg aagcccttcg agagagacat ctccacagaa 600 atctaccagg ccggcagcac cccttgcaat ggcgtggagg gctttaactg ttatttcccc 660 ctccagtcct acggctttca gcctaccaac ggcgtgggct atcagccata ccgcgtggtg 720 gtgctgtcct tcgagctgct gcacgctcca gctacagt gt gcggccctaa gaagagcacc 780 aatctggtga agaacaagtg cgtgaacttc aacttcaacg gcctgaccgg cacaggcgtg 840 ctgaccgagt ccaataagaa gttcctgccc tttcagcagt tcggcagaga catcgccgat 900 accacagacg ctgtgcgcga tccccagacc ctggagatcc tggacatcac accttgcagc 960 tttggcggcg tgtctgtgat cacacctggc accaatacaa gcaaccaggt ggccgtgctg 1020 tatcaggatg tgaattgtac cgaggtgcca gtggctatcc acgccgacca gctgaccccc 1080 acatggaggg tgtactccac cggcagcaac gtgttccaga cacgggccgg atgtctgatc 1140 ggagctgagc atgtgaacaa ttcctatgag tgcgacatcc caatcggcgc cggcatctgt 1200 gcctcctacc agacccagac aaacagccca aggcgggcca ggtctgtggc ttcccagagc 1260 atcatcgcct ataccatgtc cctgggcgcc ggctccggct ccggccagta catcaaggcc 1320 aactccaagt tcatcggcat caccgagctg taa 1353 <210> 19 <211> 18 <212> PRT <213> Unknown <220> <223> Human albumin signal peptide <400 > 19 Met Lys Trp Val Thr Phe Ile Ser Leu Leu Phe Leu Phe Ser Ser Ala 1 5 10 15 Tyr Ser <210> 20 <211> 15 <212> PRT <213> Unknown <220> <223> Tetanus Toxoid Epitope- P2 domain <400> 20 Gln Tyr I le Lys Ala Asn Ser Lys Phe Ile Gly Ile Thr Glu Leu 1 5 10 15 <210> 21 <211> 418 <212> PRT <213> Artificial Sequence <220> <223> N protein of SARS-CoV-2 <400 > 21 Ser Asp Asn Gly Pro Gln Asn Gln Arg Asn Ala Pro Arg Ile Thr Phe 1 5 10 15 Gly Gly Pro Ser Asp Ser Thr Gly Ser Asn Gln Asn Gly Glu Arg Ser 20 25 30 Gly Ala Arg Ser Lys Gln Arg Arg Pro Gln Gly Leu Pro Asn Asn Thr 35 40 45 Ala Ser Trp Phe Thr Ala Leu Thr Gln His Gly Lys Glu Asp Leu Lys 50 55 60 Phe Pro Arg Gly Gln Gly Val Pro Ile Asn Thr Asn Ser Ser Pro Asp 65 70 75 80 Asp Gln Ile Gly Tyr Tyr Arg Arg Ala Thr Arg Arg Ile Arg Gly Gly 85 90 95 Asp Gly Lys Met Lys Asp Leu Ser Pro Arg Trp Tyr Phe Tyr Tyr Leu 100 105 110 Gly Thr Gly Pro Glu Ala Gly Leu Pro Tyr Gly Ala Asn Lys Asp Gly 115 120 125 Ile Ile Trp Val Ala Thr Glu Gly Ala Leu Asn Thr Pro Lys Asp His 130 135 140 Ile Gly Thr Arg Asn Pro Ala Asn Asn Ala Ala I le Val Leu Gln Leu 145 150 155 160 Pro Gln Gly Thr Thr Leu Pro Lys Gly Phe Tyr Ala Glu Gly Ser Arg 165 170 175 Gly Gly Ser Gln Ala Ser Ser Arg Ser Ser Ser Arg Ser Arg Asn Ser 180 185 190 Ser Arg Asn Ser Thr Pro Gly Ser Ser Arg Gly Thr Ser Pro Ala Arg 195 200 205 Met Ala Gly Asn Gly Gly Asp Ala Ala Leu Ala Leu Leu Leu Leu Asp 210 215 220 Arg Leu Asn Gln Leu Glu Ser Lys Met Ser Gly Lys Gly Gln Gln Gln 225 230 235 240 Gln Gly Gln Thr Val Thr Lys Lys Ser Ala Ala Glu Ala Ser Lys Lys 245 250 255 Pro Arg Gln Lys Arg Thr Ala Thr Lys Ala Tyr Asn Val Thr Gln Ala 260 265 270 Phe Gly Arg Arg Gly Pro Glu Gln Thr Gln Gly Asn Phe Gly Asp Gln 275 280 285 Glu Leu Ile Arg Gln Gly Thr Asp T yr Lys His Trp Pro Gln Ile Ala 290 295 300 Gln Phe Ala Pro Ser Ala Ser Ala Phe Phe Gly Met Ser Arg Ile Gly 305 310 315 320 Met Glu Val Thr Pro Ser Gly Thr Trp Leu Thr Tyr Thr Gly Ala Ile 325 330 335 Lys Leu Asp Asp Lys Asp Pro Asn Phe Lys Asp Gln Val Ile Leu Leu 340 345 350 Asn Lys His Ile Asp Ala Tyr Lys Thr Phe Pro Pro Thr Glu Pro Lys 355 360 365 Lys Asp Lys Lys Lys Lys Ala Asp Glu Thr Gln Ala Leu Pro Gln Arg 370 375 380 Gln Lys Lys Gln Gln Thr Val Thr Leu Leu Pro Ala Ala Asp Leu Asp 385 390 395 400 Asp Phe Ser Lys Gln Leu Gln Gln Ser Met Ser Ser Ala Asp Ser Thr 405 410 415 Gln Ala <210 > 22 <211> 1311 <212> DNA <213> Artificial Sequence <220> <223> N protein of SARS-CoV-2_BEVS <400> 22 atgaaatggg tcac cttcat cagtctgctg ttcctgttct cttccgctta ctcctccgac 60 aacggtcctc aaaaccaacg caacgcaccc cgcatcacct tcggtggccc aagcgactct 120 actggttcca accagaacgg tgaacgctca ggcgctcgtt ccaagcagcg ccgtccacag 180 ggcctgccta acaacaccgc ttcctggttc accgccctga ctcagcacgg aaaggaggac 240 ctgaagttcc ctcgtggaca gggtgtgccc atcaacacca actccagccc tgacgaccag 300 atcggatact acaggagagc cactcgccgt atcaggggag gtgacggcaa gatgaaggac 360 ctgtccccca gatggtactt ctactacctc ggcaccggac ccgaggctgg actgccatac 420 ggtgccaaca aggacggtat catctgggtg gctaccgaag gcgccctgaa cactcccaag 480 gaccacatcg gtactaggaa cccagctaac aacgctgcca tcgtcctgca actgccacag 540 ggcaccactc tgcctaaggg tttctacgct gaaggcagcc gcggcggatc tcaggcctct 600 tcacgttcca gctctcgctc ccgtaactca tccaggaaca gcaccccagg cagctctagg 660 ggaacttctc ctgctagaat ggctggaaac ggtggcgacg ctgccctggc tctgctgctg 720 ctggacagac tgaaccagct ggagagcaag atgtctggca agggacagca gcagcaggga 780 cagactgtga ccaagaagtc cgctgctgag gcttccaaga agcccaggca gaagagaacc 840 gctactaagg cctacaacgt cacccaggcc ttc ggaagga gaggtccaga gcagactcag 900 ggcaacttcg gtgaccagga actgatccgc cagggcaccg actacaagca ctggcctcag 960 atcgctcagt tcgccccctc agcttccgcc ttcttcggaa tgtctcgtat cggtatggaa 1020 gtgaccccat caggcacttg gctgacctac actggagcta tcaagctgga tgacaaggac 1080 cctaacttca aggaccaggt catcctgctg aacaagcaca tcgacgccta caagaccttc 1140 cctcccactg agcctaagaa ggacaagaag aagaaggctg acgaaaccca ggccctgcct 1200 cagcgccaga agaagcagca gactgtcact ctgctgcccg ctgccgacct ggacgacttc 1260 agcaagcagc tgcaacagtc tatgtcatcc gctgactcaa ctcaggccta a 1311 <210> 23 <211> 1311 <212> DNA <213> Artificial Sequence <220> <223> N protein of SARS-CoV-2_CHO <400> 23 atgaagtggg tcactttcat cagcctgttg tttctgttca gctccgctgccta ctagacttcagat ccatccgccta ctagacttcagat 60 caccagtccacc t aagcgactca 120 acaggcagta accagaacgg cgagcggtcc ggcgctagat ccaagcagag acggcctcag 180 ggcctgccaa acaacaccgc ctcttggttt accgctctga cccagcacgg caaggaggtc accg caaggatagaggtc acc cc cc cagcaataga acctc acc cc aaggaga atccgcggcg gcgacggcaa gatgaaggac 360 ctgtccccac ggtggtactt ctactatctg ggcaccggcc ccgaggctgg cctgccttat 420 ggcgctaaca aggatggcat catctgggtg gctacagagg gcgctctgaa tacccctaag 480 gatcacatcg gcacaagaaa tccagctaat aacgccgcta tcgtgctgca actgccccag 540 ggcaccacac tgccaaaggg cttttacgct gagggctctc gcggcggctc ccaggcttct 600 tccagaagct cttccagatc cagaaactcc tctcgcaact ctacccctgg ctcttccaga 660 ggcacaagcc ctgctagaat ggccggcaat ggcggcgacg ccgctctggc cctgctgctg 720 ctggataggc tgaaccagct ggagtccaag atgtctggca agggccagca gcagcagggc 780 cagacagtga ccaagaagtc tgccgctgag gcttccaaga agcctcggca gaagagaacc 840 gccacaaagg cttataacgt gacccaggct tttggcagaa gaggccctga gcagacccag 900 ggcaacttcg gcgatcagga gctgatcaga cagggcaccg attacaagca ttggccacag 960 atcgcccagt ttgctccttc cgccagcgcc ttctttggca tgtccaggat cggcatggag 1020 gtgacaccct ctggcacctg gctgacatat accggcgcta tcaagctgga cgataaggac 1080 ccaaacttca aggatcaggt aatcctgctg aacaagcaca tcgacgccta caagacattc 1140 ccacctaccg agccaaagaa ggacaagaag aagaaggcc g atgaaaccca ggccctgccc 1200 cagagacaga agaagcagca gacagtgacc ctgctgccag ctgccgatct ggacgatttc 1260 tcaaaacagc ttcagcagtc aatgtcatcc gccgattcaa ctcaggcata a Nucleus Sequence Coding for the Gene Sequence <220> 223 <211> Artificial Sequence <220> spike of 223 <211> protein of SARS-CoV-2 <400> 24 atgtttgttt ttcttgtttt attgccacta gtctctagtc agtgtgttaa tcttacaacc 60 agaactcaat taccccctgc atacactaat tctttcacac gtggtgttta ttaccctgac 120 aaagttttca gatcctcagt tttacattca actcaggact tgttcttacc tttcttttcc 180 aatgttactt ggttccatgc tatacatgtc tctgggacca atggtactaa gaggtttgat 240 aaccctgtcc taccatttaa tgatggtgtt tattttgctt ccactgagaa gtctaacata 300 ataagaggct ggatttttgg tactacttta gattcgaaga cccagtccct acttattgtt 360 aataacgcta ctaatgttgt tattaaagtc tgtgaatttc aattttgtaa tgatccattt 420 ttgggtgttt attaccacaa aaacaacaaa agttggatgg aaagtgagtt cagagtttat 480 tctagtgcga ataattgcac ttttgaatat gtctctcagc cttttcttat ggaccttgaa 540 ggaaaacagg gtaatttcaa aaatcttagg gaatttgtgt ttaagaatat tg atggttat 600 tttaaaatat attctaagca cacgcctatt aatttagtgc gtgatctccc tcagggtttt 660 tcggctttag aaccattggt agatttgcca ataggtatta acatcactag gtttcaaact 720 ttacttgctt tacatagaag ttatttgact cctggtgatt cttcttcagg ttggacagct 780 ggtgctgcag cttattatgt gggttatctt caacctagga cttttctatt aaaatataat 840 gaaaatggaa ccattacaga tgctgtagac tgtgcacttg accctctctc agaaacaaag 900 tgtacgttga aatccttcac tgtagaaaaa ggaatctatc aaacttctaa ctttagagtc 960 caaccaacag aatctattgt tagatttcct aatattacaa acttgtgccc ttttggtgaa 1020 gtttttaacg ccaccagatt tgcatctgtt tatgcttgga acaggaagag aatcagcaac 1080 tgtgttgctg attattctgt cctatataat tccgcatcat tttccacttt taagtgttat 1140 ggagtgtctc ctactaaatt aaatgatctc tgctttacta atgtctatgc agattcattt 1200 gtaattagag gtgatgaagt cagacaaatc gctccagggc aaactggaaa gattgctgat 1260 tataattata aattaccaga tgattttaca ggctgcgtta tagcttggaa ttctaacaat 1320 cttgattcta aggttggtgg taattataat tacctgtata gattgtttag gaagtctaat 1380 ctcaaacctt ttgagagaga tatttcaact gaaatctatc aggccggtag cacaccttgt 1440 aatggtgttg aaggttttaa ttgttacttt cctttacaat catatggttt ccaacccact 1500 aatggtgttg gttaccaacc atacagagta gtagtacttt cttttgaact tctacatgca 1560 ccagcaactg tttgtggacc taaaaagtct actaatttgg ttaaaaacaa atgtgtcaat 1620 ttcaacttca atggtttaac aggcacaggt gttcttactg agtctaacaa aaagtttctg 1680 cctttccaac aatttggcag agacattgct gacactactg atgctgtccg tgatccacag 1740 acacttgaga ttcttgacat tacaccatgt tcttttggtg gtgtcagtgt tataacacca 1800 ggaacaaata cttctaacca ggttgctgtt ctttatcagg atgttaactg cacagaagtc 1860 cctgttgcta ttcatgcaga tcaacttact cctacttggc gtgtttattc tacaggttct 1920 aatgtttttc aaacacgtgc aggctgttta ataggggctg aacatgtcaa caactcatat 1980 gagtgtgaca tacccattgg tgcaggtata tgcgctagtt atcagactca gactaattct 2040 cctcggcggg cacgtagtgt agctagtcaa tccatcattg cctacactat gtcacttggt 2100 gcagaaaatt cagttgctta ctctaataac tctattgcca tacccacaaa ttttactatt 2160 agtgttacca cagaaattct accagtgtct atgaccaaga catcagtaga ttgtacaatg 2220 tacatttgtg gtgattcaac tgaatgcagc aatcttttgt tgcaatatgg cagtttttgt 2280 acaca attaa accgtgcttt aactggaata gctgttgaac aagacaaaaa cacccaagaa 2340 gtttttgcac aagtcaaaca aatttacaaa acaccaccaa ttaaagattt tggtggtttt 2400 aatttttcac aaatattacc agatccatca aaaccaagca agaggtcatt tattgaagat 2460 ctacttttca acaaagtgac acttgcagat gctggcttca tcaaacaata tggtgattgc 2520 cttggtgata ttgctgctag agacctcatt tgtgcacaaa agtttaacgg ccttactgtt 2580 ttgccacctt tgctcacaga tgaaatgatt gctcaataca cttctgcact gttagcgggt 2640 acaatcactt ctggttggac ctttggtgca ggtgctgcat tacaaatacc atttgctatg 2700 caaatggctt ataggtttaa tggtattgga gttacacaga atgttctcta tgagaaccaa 2760 aaattgattg ccaaccaatt taatagtgct attggcaaaa ttcaagactc actttcttcc 2820 acagcaagtg cacttggaaa acttcaagat gtggtcaacc aaaatgcaca agctttaaac 2880 acgcttgtta aacaacttag ctccaatttt ggtgcaattt caagtgtttt aaatgatatc 2940 ctttcacgtc ttgacaaagt tgaggctgaa gtgcaaattg ataggttgat cacaggcaga 3000 cttcaaagtt tgcagacata tgtgactcaa caattaatta gagctgcaga aatcagagct 3060 tctgctaatc ttgctgctac taaaatgtca gagtgtgtac ttggacaatc aaaaagagtt 3120 gatttttgtg gaaagggcta tcatcttatg tccttccctc agtcagcacc tcatggtgta 3180 gtcttcttgc atgtgactta tgtccctgca caagaaaaga acttcacaac tgctcctgcc 3240 atttgtcatg atggaaaagc acactttcct cgtgaaggtg tctttgtttc aaatggcaca 3300 cactggtttg taacacaaag gaatttttat gaaccacaaa tcattactac agacaacaca 3360 tttgtgtctg gtaactgtga tgttgtaata ggaattgtca acaacacagt ttatgatcct 3420 ttgcaacctg aattagactc attcaaggag gagttagata aatattttaa gaatcataca 3480 tcaccagatg ttgatttagg tgacatctct ggcattaatg cttcagttgt aaacattcaa 3540 aaagaaattg accgcctcaa tgaggttgcc aagaatttaa atgaatctct catcgatctc 3600 caagaacttg gaaagtatga gcagtatata aaatggccat ggtacatttg gctaggtttt 3660 atagctggct tgattgccat agtaatggtg acaattatgc tttgctgtat gaccagttgc 3720 tgtagttgtc tcaagggctg ttgttcttgt ggatcctgct gcaaatttga tgaagacgac 3780 tctgagccag tgctcaaagg agtcaaatta cattacacat aa 3822 <210> 25 <211> 2058 <212> DNA <213> Artificial Sequence < 220> <223> Nucleotide sequence of the gene coding for the S1 domain of spike protein of SARS-CoV-2 <400> 25 atgtttgttt tt cttgtttt attgccacta gtctctagtc agtgtgttaa tcttacaacc 60 agaactcaat taccccctgc atacactaat tctttcacac gtggtgttta ttaccctgac 120 aaagttttca gatcctcagt tttacattca actcaggact tgttcttacc tttcttttcc 180 aatgttactt ggttccatgc tatacatgtc tctgggacca atggtactaa gaggtttgat 240 aaccctgtcc taccatttaa tgatggtgtt tattttgctt ccactgagaa gtctaacata 300 ataagaggct ggatttttgg tactacttta gattcgaaga cccagtccct acttattgtt 360 aataacgcta ctaatgttgt tattaaagtc tgtgaatttc aattttgtaa tgatccattt 420 ttgggtgttt attaccacaa aaacaacaaa agttggatgg aaagtgagtt cagagtttat 480 tctagtgcga ataattgcac ttttgaatat gtctctcagc cttttcttat ggaccttgaa 540 ggaaaacagg gtaatttcaa aaatcttagg gaatttgtgt ttaagaatat tgatggttat 600 tttaaaatat attctaagca cacgcctatt aatttagtgc gtgatctccc tcagggtttt 660 tcggctttag aaccattggt agatttgcca ataggtatta acatcactag gtttcaaact 720 ttacttgctt tacatagaag ttatttgact cctggtgatt cttcttcagg ttggacagct 780 ggtgctgcag cttattatgt gggttatctt caacctagga cttttctatt aaaatataat 840 gaaaatggaa ccattacaga tgctgtagac t gtgcacttg accctctctc agaaacaaag 900 tgtacgttga aatccttcac tgtagaaaaa ggaatctatc aaacttctaa ctttagagtc 960 caaccaacag aatctattgt tagatttcct aatattacaa acttgtgccc ttttggtgaa 1020 gtttttaacg ccaccagatt tgcatctgtt tatgcttgga acaggaagag aatcagcaac 1080 tgtgttgctg attattctgt cctatataat tccgcatcat tttccacttt taagtgttat 1140 ggagtgtctc ctactaaatt aaatgatctc tgctttacta atgtctatgc agattcattt 1200 gtaattagag gtgatgaagt cagacaaatc gctccagggc aaactggaaa gattgctgat 1260 tataattata aattaccaga tgattttaca ggctgcgtta tagcttggaa ttctaacaat 1320 cttgattcta aggttggtgg taattataat tacctgtata gattgtttag gaagtctaat 1380 ctcaaacctt ttgagagaga tatttcaact gaaatctatc aggccggtag cacaccttgt 1440 aatggtgttg aaggttttaa ttgttacttt cctttacaat catatggttt ccaacccact 1500 aatggtgttg gttaccaacc atacagagta gtagtacttt cttttgaact tctacatgca 1560 ccagcaactg tttgtggacc taaaaagtct actaatttgg ttaaaaacaa atgtgtcaat 1620 ttcaacttca atggtttaac aggcacaggt gttcttactg agtctaacaa aaagtttctg 1680 cctttccaac aatttggcag agacattgct gacactact g atgctgtccg tgatccacag 1740 acacttgaga ttcttgacat tacaccatgt tcttttggtg gtgtcagtgt tataacacca 1800 ggaacaaata cttctaacca ggttgctgtt ctttatcagg atgttaactg caccatt tgacttgt c t ggt t t gacttgt c t gg t t t gactt t caact aatgtttttc aaacacgtgc aggctgttta ataggggctg aacatgtcaa caactcatat 1980 gagtgtgaca tacccattgg tgcaggtata tgcgctagtt atcagactca gactaattct the 2040 cctcggc Sequenceggg domain cacg223> Artificial of <2058 <212> coding the gene for gene <26>211> <212> of spike protein of SARS-CoV-2 <400> 26 gtagctagtc aatccatcat tgcctacact atgtcacttg gtgcagaaaa ttcagttgct 60 tactctaata actctattgc catacccaca aattttacta ttagtgttac cacagaaatt 120 ctaccagtgt ctatgaccaa gacatcagta gattgtacaa tgtacatttg tggtgattca 180 actgaatgca gcaatctttt gttgcaatat ggcagttttt gtacacaatt aaaccgtgct 240 ttaactggaa tagctgttga acaagacaaa aacacccaag aagtttttgc acaagtcaaa 300 caaatttaca aaacaccacc aattaaagat tttggtggtt ttaatttttc acaaatatta 360 ccagatccat caaaaccaag caagaggtca tttattgaag atctactttt caacaaagtg 420 acacttgcag atgctggctt catcaaacaa tatggtgatt gccttggtga tattgctgct 480 agagacctca tttgtgcaca aaagtttaac ggccttactg ttttgccacc tttgctcaca 540 gatgaaatga ttgctcaata cacttctgca ctgtta gcgg gtacaatcac ttctggttgg 600 acctttggtg caggtgctgc attacaaata ccatttgcta tgcaaatggc ttataggttt 660 aatggtattg gagttacaca gaatgttctc tatgagaacc aaaaattgat tgccaaccaa 720 tttaatagtg ctattggcaa aattcaagac tcactttctt ccacagcaag tgcacttgga 780 aaacttcaag atgtggtcaa ccaaaatgca caagctttaa acacgcttgt taaacaactt 840 agctccaatt ttggtgcaat ttcaagtgtt ttaaatgata tcctttcacg tcttgacaaa 900 gttgaggctg aagtgcaaat tgataggttg atcacaggca gacttcaaag tttgcagaca 960 tatgtgactc aacaattaat tagagctgca gaaatcagag cttctgctaa tcttgctgct 1020 actaaaatgt cagagtgtgt acttggacaa tcaaaaagag ttgatttttg tggaaagggc 1080 tatcatctta tgtccttccc tcagtcagca cctcatggtg tagtcttctt gcatgtgact 1140 tatgtccctg cacaagaaaa gaacttcaca actgctcctg ccatttgtca tgatggaaaa 1200 gcacactttc ctcgtgaagg tgtctttgtt tcaaatggca cacactggtt tgtaacacaa 1260 aggaattttt atgaaccaca aatcattact acagacaaca catttgtgtc tggtaactgt 1320 gatgttgtaa taggaattgt caacaacaca gtttatgatc ctttgcaacc tgaattagac 1380 tcattcaagg aggagttaga taaatatttt aagaatcata catcacca ga tgttgattta 1440 ggtgacatct ctggcattaa tgcttcagtt gtaaacattc aaaaagaaat tgaccgcctc 1500 aatgaggttg ccaagaattt aaatgaatct ctcatcgatc tccaagaact tggaaagtat 1560 gagcagtata taaaatggcc atggtacatt tggctaggtt ttatagctgg cttgattgcc 1620 atagtaatgg tgacaattat gctttgctgt atgaccagtt gctgtagttg tctcaagggc 1680 tgttgttctt gtggatcctg ctgcaaattt gatgaagacg actctgagcc agtgctcaaa 1740 ggagtcaaat tacattacac ataa 1764 <210> 27 <211> 582 <212> DNA <213> Artificial Sequence <220> <223> Nucleotide sequence of the gene coding for the RBD of spike protein of SARS-CoV-2 <400> 27 aatattacaa acttgtgccc ttttggtgaa gtttttaacg ccaccagatt tgcatctgtttt 60 tatgctttgct g attagataact t a 120 tccgcatcat tttccacttt taagtgttat ggagtgtctc ctactaaatt aaatgatctc 180 tgctttacta atgtctatgc agattcattt gtaattagag gtgatgaagt cagacaaatc 240 gctccagggc aaactggaaa gattgctgat tataattata aattaccaga tgattttaca 300 ggctgcgtta tagcttggaa ttctaacaat cttgattcta aggttggtgg taattataat 360 tacctgtata gattgt ttag gaagtctaat ctcaaacctt ttgagagaga tatttcaact 420 gaaatctatc aggccggtag cacaccttgt aatggtgttg aaggttttaa ttgttacttt 480 cctttacaat catatggttt ccaacccact aatggtgttg gttaccaacc atacagagta 540 gtagtacttt cttttgaact tctacatgca ccagcaactg tt 582 <210> 28 <211> 1274 <212> PRT <213> Unknown <220> <223> Amino acid sequence of the spike protein of SARS-CoV-2 <400> 28 Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val 1 5 10 15 Asn Leu Thr 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 Leu 1010 1015 1020 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val 1025 1030 1035 1040 Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala 1045 1050 1055 Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu 1060 1065 1070 Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His 1075 1080 1085 Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val 1090 1095 1100 Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr 1105 1110 1115 1120 Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr 1125 1130 1135 Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu 1140 1145 1150 Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp 1155 1160 1165 Ile Ser Gly Ile Asn Ala Ser Val Va l Asn Ile Gln Lys Glu Ile Asp 1170 1175 1180 Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1185 1190 1195 1200 Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile 1205 1210 1215 Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile 1220 1225 1230 Met 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 Val 1250 1255 1260 Leu Lys Gly Val Lys Leu His Tyr Thr *** 1265 1270 <210> 29 <211> 673 <212> PRT <213> Unknown <220> <223> Amino acid sequence of the S1 domain of spike protein of SARS-CoV-2 <400> 29 Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr 1 5 10 15 Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser 20 25 30 Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn 35 40 45 Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys 50 55 60 Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala 65 70 75 80 Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr 85 90 95 Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn 100 105 110 Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu 115 120 125 Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe 130 135 140 Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln 145 150 155 160 Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu 165 170 175 Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser 180 185 190 Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser 195 200 205 Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg 210 215 220 Phe Gln Thr Leu Leu Ala Leu His A rg Ser Tyr Leu Thr Pro Gly Asp 225 230 235 240 Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr 245 250 255 Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile 260 265 270 Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys 275 280 285 Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn 290 295 300 Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr 305 310 315 320 Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser 325 330 335 Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr 340 345 350 Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly 355 360 365 Val Ser Pro Thr Lys L eu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala 370 375 380 Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly 385 390 395 400 Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe 405 410 415 Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val 420 425 430 Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu 435 440 445 Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser 450 455 460 Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln 465 470 475 480 Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg 485 490 495 Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys 500 505 510 Gly Pro L ys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe 515 520 525 Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys 530 535 540 Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr 545 550 555 560 Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro 565 570 575 Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser 580 585 590 Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro 595 600 605 Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser 610 615 620 Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala 625 630 635 640 Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly 645 650 6 55 Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg 660 665 670 Ser <210> 30 <211> 588 <212> PRT <213> Unknown <220> <223> Amino acid sequence of the S2 domain of spike protein of SARS-CoV-2 <400> 30 Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu 1 5 10 15 Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe 20 25 30 Thr Ile Ser Val Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr 35 40 45 Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser 50 55 60 Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala 65 70 75 80 Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe 85 90 95 Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Ile Lys Asp Phe Gly 100 105 110 Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys 115 120 125 Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp 130 135 140 Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala 145 150 155 160 Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro 165 170 175 Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu 180 185 190 Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu 195 200 205 Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly 210 215 220 Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln 225 230 235 240 Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala 245 250 255 Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala 260 265 270 Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser 275 280 285 Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu 290 295 300 Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr 305 310 315 320 Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala 325 330 335 Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys 340 345 350 Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 355 360 365 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala 370 375 380 Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys 385 390 395 400 Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp 405 410 415 Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp 420 425 430 Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn 435 440 445 Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu 450 455 460 Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu 465 470 475 480 Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu 485 490 495 Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile 500 505 510 Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp 515 520 525 Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 530 535 540 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly 545 550 555 560 Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu 565 570 575 Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr ** * 580 585 <210> 31 <211> 194 <212> PRT <213> Unknown <220> <223> Amino acid sequence of the RBD of spike protein of SARS-CoV-2 <400> 31 Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg 1 5 10 15 Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val 20 25 30 Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys 35 40 45 Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn 50 55 60 Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile 65 70 75 80 Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro 85 90 95 Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp 100 105 110 Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys 115 120 125 Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln 130 135 140 Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe 145 150 155 160 Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln 165 170 175 Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala 180 185 190Thr Val

Claims (31)

It is a recombinant antigen protein derived from the spike protein (S protein) of SARS-coronavirus-2, in which all or part of the S2 subunit of the spike protein has been removed,
The recombinant antigen protein is SARS- Recombinant antigen protein, characterized in that the antigen for the prevention or treatment of coronavirus-2 infection. The recombinant antigen protein according to claim 1, wherein the entire S2 subunit is removed from the spike protein, and the recombinant antigen protein is represented by SEQ ID NO: 1. According to claim 2, wherein the recombinant antigen protein is characterized in that the four amino acids corresponding to positions 683 to 686 of the spike protein are further removed, the recombinant antigen protein. The recombinant antigen protein according to any one of claims 1 to 3, characterized in that a P2 domain is further linked to the C terminus of the recombinant antigen protein. The method of claim 1, wherein the recombinant antigen protein is
A recombinant antigen protein characterized in that a part of the N-terminal domain of the S1 subunit of the SARS-coronavirus-2 spike protein has been removed, and the N-terminal part of the S2 subunit is included at the C-terminus. According to claim 5, wherein the recombinant antigen protein is characterized in that consisting of the amino acid sequence of positions 290 to 701 of the S protein, the recombinant antigen protein. The recombinant antigen protein according to claim 6, characterized in that the P2 domain is further linked to the C-terminus of the recombinant antigen protein. 5. The recombinant antigen protein according to claim 4, wherein the P2 domain is linked by a GSGSG linker. The recombinant antigen protein according to claim 1, wherein the recombinant antigen protein has any one sequence selected from the group consisting of SEQ ID NOs: 1 to 6. Claims 1 to 3, claim 5 to claim 7, and claim 9, comprising the recombinant antigen protein according to any one of claims 9, SARS-coronavirus-2 prophylactic or therapeutic antigen composition. As an expression construct for the production of recombinant antigen protein for the prevention or treatment of SARS-coronavirus-2 infection,
An expression construct comprising a nucleic acid sequence encoding a peptide in which all or part of the S2 subunit of the SARS-coronavirus-2 spike protein (S protein) has been removed. The method according to claim 11, wherein the expression construct encodes a signal peptide derived from itself or heterologous to a nucleic acid sequence encoding a peptide in which all or part of the S2 subunit of the SARS-coronavirus-2 spike protein (S protein) has been removed. An expression construct in which a nucleic acid sequence is further fused. The expression construct according to claim 12, wherein the heterologous signal peptide is a human albumin signal peptide. The expression construct according to claim 13, wherein the expression construct is further fused with a nucleic acid sequence encoding the P2 domain of Tetanus toxin. The expression construct according to claim 11, wherein the expression construct has any one nucleic acid sequence selected from SEQ ID NOs: 7 to 18. A vector comprising the expression construct according to any one of claims 11 to 15. A host cell having the vector of claim 16 . A vector comprising the expression construct according to any one of claims 11 to 15. A host cell having the vector of claim 18 . The host cell of claim 19, wherein the host cell is a Chinese hamster ovary (CHO) cell, and the expression construct has any one nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13 to 18. A baculovirus recombinant vector comprising the expression construct according to any one of claims 11 to 15. The baculovirus recombinant vector according to claim 21, wherein the expression construct comprises any one expression construct selected from the group consisting of SEQ ID NOs: 7 to 12. A recombinant Bacmid obtained by transforming the baculovirus recombinant vector according to claim 22 into E. coli. A recombinant baculovirus comprising the recombinant vacmid according to claim 22 . A host cell comprising the recombinant bacmid according to claim 22 . The host cell according to claim 25, wherein the host cell is an insect cell comprising Sf21 or Sf9. A method for producing a recombinant antigenic protein comprising culturing the host cell of claim 19 or 25 and isolating the desired product. Use of the antigenic protein according to any one of claims 1 to 3, 5 to 7 and 9 for the prevention or treatment of SARS-coronavirus-2 infection. SARS-coronavirus- for preventing or treating SARS-coronavirus-2 infection by administering the antigenic protein according to any one of claims 1 to 3, 5 to 7 and 9 to an individual 2 How to prevent or treat an infection. Claims 1 to 3, claim 5 to claim 7, comprising the antigen protein according to any one of claims 9 and a pharmaceutically acceptable carrier, SARS - for preventing or treating coronavirus-2 infection vaccine composition. 31. The method of claim 30, wherein the composition comprises
SEQ ID NO: 21 SARS-coronavirus-2 nucleocapsid (N) protein, an immunological adjuvant or a mixture thereof, further comprising a SARS-coronavirus-2 infection prevention or treatment vaccine composition.

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