TW201249996A - Novel vaccines against pandemic influenza virus A/H1N1 - Google Patents

Abstract

The invention relates to influenza virus vaccines, and in particular to an influenza virus which has at least its hemagglutinin gene comprising a D222 mutation, and an oseltamivir resistant neuraminidase gene. In some embodiments the influenza virus vaccine further comprises a matrix protein comprising a mutation that increases specific immune response in humans. The virus is useful for production of vaccines against hypervirulent and oseltamivir-resistant influenza.

Description

201249996 6. INSTRUCTIONS OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to anti-influenza virus vaccines, and in particular to epidemic areas of supervirulent strains against pandemic A/H1N1 influenza virus. The present invention is useful for the production of influenza virus strains that carry influenza vaccines with specific mutations in the A and NA segments. The present application claims priority to U.S. Provisional Patent Application Serial No. 61,409/303, filed on Nov. 2, 2010. This provisional application is incorporated herein in its entirety by reference. [Prior Art] Influenza is caused by the RNA virus of the genus rth〇myx〇viridae. There are three types of these viruses, and they cause three different types of influenza: sputum type, sputum type and Type c. Influenza-type influenza virus infects mammals (humans, pigs, ferrets, horses) and birds. This is extremely important to humans because this type of virus has caused widespread disease worldwide. Influenza virus (also referred to as influenza (1) only infects humans. It sometimes causes a local outbreak of influenza. Influenza prions only infect humans. They have the highest number of infections in the early stages and rarely cause serious diseases. The cold A virus infects a wide variety of mammals, including humans, horses, pigs, cedar and birds. Major human pathogens are associated with epidemics and pandemics. There are at least 15 known hemagglutinin (H) serotypes and 9 The neuraminidase (N) serotype is known. It is believed that pigs and birds are particularly important reservoirs that generate genetically and antigenically distinct virus pools that are 15989 0.doc 201249996 Transferred back to the population by close contact between humans and animals. Influenza B virus only infects mammals and causes disease, but is generally less severe than type A. Unlike influenza A virus, influenza B virus There is no discernible serotype. Influenza C virus only infects mammals, but extremely; &, causes disease. It is genetically and morphologically different from type A and type b. Unlike many other viruses, it is popular. The genomic genome consists of RNA rather than DNA. The genome contains 8 segments and only contains viral particles of all 8 segments. (Steinhauer DA, Skehel π, Genetics of influenza viruses. Ann Rev Genet. 2002 36: 305-332). There are four antigens in the influenza virus: hemagglutinin (HA), neuraminidase (ΝΑ), nucleocapsid (NA), matrix (M) and nucleocapsid protein (NP). ° NP is a type-specific antigen and is stored in three forms of A B&c

In, it knows: the basis for the classification of human influenza virus. The matrix protein (M protein) surrounds the nucleocapsid and accounts for 35-45% of the mass of the particles. Two surface glycoproteins were observed to have rod-like projections on the surface. Hemagglutinin (HA) consists of two human units, HA1 and HA2. HA mediates the attachment of the virus to cellular receptors. The neurokinase (NA) molecule is present in smaller amounts in the mantle. Circulating human strains are well known for their frequent accumulation of mutations year after year and the recurrence of epidemics. The influenza virus is one of the few viruses in a single segment (8). The segmentation of the genome also allows for the exchange of complete genes between different strains, thereby increasing the likelihood of recombinant formation (if two different viruses infect the same cell, they can be formed by swapping the gene segments). That is, if a cell is infected by two different influenza strains, each segment 159890.doc 201249996 can be redistributed to produce a new virus with a segment from each initial virus. In nature, this can help to quickly produce new influenza strains. A new genome can be assembled by mixing between the two viruses. This process can also be replicated in the laboratory (for the manufacture of vaccine strains). In the Far East, poultry and human strains that are reconstituted in pigs allow the evolution of virulent human strains. The genetic plasticity of influenza viruses is also likely to be related to vaccine design, pathogenicity, and the ability of new viruses to emerge from natural repositories and cause worldwide pandemics. The pandemic (H1N1) 2009 virus has evolved worldwide, and it has dominated from the initial mixed branching pattern to a branching (branch 7) during the pandemic process. Therefore, the virus that makes up this branch can be responsible for most of the pandemic that has broken out worldwide. The mutation S203T in the globulin (HA) protein sequence is unique to the branch 7 isolate. (Valli MB et al, Evolutionary pattern of pandemic influenza (H1N1) 2009 virus in the late phases of the 2009 pandemic. PLoS Curr Influenza. March 3, 2010: RRN1149) o In the branch 7 isolate containing the S203T mutation A change in the amino acid corresponding to position 222 (position 225 of the H3 number) in the hemagglutinin HA1 gene sequence (in which aspartic acid (D) is changed to glycine (G)) is associated with severe clinical outcome. D222G mutations were recorded in the H1N1 pdm global isolate in the GenBank at various time points between April and November 2009 (Chen GW, Shih SR (2009) Genomic signatures of influenza A Pandemic (H1N1) 2009 virus. Emerg Infect Dis). 159890.doc 201249996 The influenza vaccine is an annual vaccine against the protection of highly susceptible influenza viruses. Each seasonal influenza vaccine injected contains three influenza viruses: an A (H3N2) virus, a 2009 prevalent H1N1 virus, and an influenza B virus. (www.cdc.gov/vaccines/pubs/vis/downloads/vis-flu.pdf). However, the pandemic ten influenza vaccine is only marginally effective against the influenza H1N1 virus carrying the D222G mutation in the HA protein. Therefore, there is an urgent need in the art to generate a virus strain and a virus-derived antigen which are robust against the potency of the D222G variant of H1N1. SUMMARY OF THE INVENTION The present invention relates to methods and compositions for the preparation of vaccines against the hypervirulent form of the pandemic influenza A H1N1 virus and/or the oseltamivir resistant form. A vaccine composition comprising a mutation in the D222 residue of HA and the S275 residue of NA is provided. The present invention provides an influenza virus vaccine comprising at least the following influenza virus: a hemagglutinin (HA) gene segment comprising a virus 1 derived from a pandemic influenza A H1/N1 influenza virus Amino acid sequence of SEQ ID NO: 34; and a neuraminidase (NA) gene segment comprising an amino acid sequence derived from a virus 1 isolate of influenza A H1/N1 influenza virus SEQ ID NO: 39. In one embodiment, the HA gene segment comprises an amino acid sequence encoded by the nucleotide sequence of segment 4 of virus 1 (SEQ ID NO: 4). In some forms, the HA gene segment comprises a branched 7-specific HA sequence of pandemic influenza A H1/N1 influenza virus. 159890.doc 201249996 In one embodiment, the HA gene segment comprises a mutation at the D222 residue in the wild-type pandemic influenza A H1/N1 influenza virus. In one embodiment, the HA gene segment comprises a D222E mutation. In some embodiments, the HA gene segment comprises a S203T mutation relative to a wild-type pandemic influenza A H1/N1 influenza virus. The present invention relates to vaccines wherein the HA gene segment further comprises one or more of the P83S and 132IV mutations. In one embodiment, the NA gene segment comprises an amino acid sequence encoded by the nucleotide sequence of segment 6 of virus 1 (SEQ ID NO: 6). The NA gene segment contains the Y275H mutation relative to the wild-type pandemic influenza A H1/N1 influenza virus. The present invention relates to a vaccine wherein the NA gene segment further comprises one or more of a TION 06V mutation, a D248N mutation. The vaccine of the present invention may further comprise: c) a matrix (MA) gene segment encoding an amino acid sequence comprising the virus 1 isolate derived from the pandemic influenza A H1/N1 influenza virus SEQ ID NO: 50 Ml peptide. In one embodiment, the ΜΑ gene segment comprises an amino acid sequence encoded by the nucleotide sequence of segment 7 of virus 1 (SEQ ID NO: 7). In some embodiments, the matrix (MA) gene segment encodes an M2 peptide comprising the amino acid sequence SEQ ID NO: 51 derived from the virus 1 isolate of the pandemic influenza A H1/N1 influenza virus. In some embodiments, the Ml peptide comprises an A198P mutation. The present invention relates to a method of immunizing a patient against an influenza virus, which comprises the step of administering the influenza vaccine disclosed herein to a patient. 159890.doc 201249996 The present invention relates to a method for immunizing a patient against hypertoxic influenza epidemics' which comprises the step of administering the influenza vaccine disclosed herein to a patient. The present invention relates to a method of immunizing a patient against oseltamivir-resistant influenza virus, which comprises the step of administering the influenza vaccine disclosed herein to a patient. The present invention is directed to a vaccine as disclosed herein, wherein the vaccine is a redistribution vaccine. In some embodiments, a redistribution influenza virus is obtained by conventional redistribution. The present invention relates to vaccines disclosed herein which are live attenuated vaccines or inactivated vaccines. In some embodiments, the vaccine is formulated for oral or intranasal administration. The present invention relates to a method of preparing a vaccine for immunizing an individual against a hypervirulent influenza virus strain comprising the steps of admixing the influenza virus disclosed herein with a carrier and optionally an adjuvant. The changes and modifications may be made without departing from the spirit and scope of the novel concept of the present disclosure. [Embodiment] The following drawings form a part of this specification and are included to further illustrate some aspects of the present disclosure, with reference to one or more of the drawings and the detailed description of the specific embodiments described herein, The invention can be understood more thoroughly. This patent or application file contains at least one color schema. A copy of the I59890.doc 201249996 patent or patent application publication with a color map will be provided by the Patent Office upon request and payment of the necessary fee. In the context of the present invention and in the specific context in which each term is used, the terms used in this specification generally have the ordinary meaning in the art. Certain terms used to describe the invention are discussed below or elsewhere in the specification, which provides additional guidance to those of ordinary skill in the art. For convenience, some terms may be highlighted, for example, in italics and/or quotation marks. The use of highlighting has no effect on the scope and meaning of the term; in the same context, the scope and meaning of the term are the same whether highlighted or not. It should be understood that the same object can be described in more than one way. Thus, alternative language and synonyms may be used in any one or more of the terms discussed herein, regardless of whether the terms are specifically recited or discussed herein without any particular meaning. Provide synonyms for certain terms. The enumeration of one or more synonyms does not exclude the use of other synonyms. The use of any of the examples in this specification, including the examples of any of the terms discussed herein, is merely illustrative and is in no way intended to limit the scope and meaning of the invention. Also, the present invention is not limited to the various embodiments given in the specification. All technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the invention pertains, unless otherwise claimed. In the event of a conflict, the present document (including definitions) shall prevail. The following references provide the skilled person with a general definition of many terms used in the present invention. Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed., 1993); The Cambridge Dictionary of Science and Technology (Walker Editor, Cambridge University 159890.doc 201249996

Press·1990); The Glossary of Genetics, 5th ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Margham, The Harper Collins Dictionary of Biology (1991). In general, the terms and techniques associated with the cell and tissue culture, molecular biology, and protein and oligonucleotide or polynucleotide chemistry and hybridization described herein are well known and commonly employed in the art. Recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipofection) use standard techniques. Enzymatic reactions and purification techniques are carried out in accordance with the manufacturer's instructions or as commonly used in the art or as described herein. Unless expressly stated to the contrary, the present invention will employ the methods of virology, immunology, microbiology, molecular biology, and recombinant DNA techniques well known in the art, many of which are described for illustrative purposes. Below. These techniques are fully explained in the literature. See, for example, Sambrook et al, Molecular Cloning: A Laboratory Manual (2nd ed., 1989); Maniatis et al, Molecular Cloning: A Laboratory Manual (1982); DNA Cloning: A Practical Approach, Volumes I and II ( Edited by D. Glover); Oligonucleotide Synthesis (edited by N. Gait, 1984); Nucleic Acid Hybridization (edited by B. Hames and S. Higgins, 1985); Transcription and Translation (edited by B. Hames and S. Higgins, 1984); Cell Culture (edited by R. Freshney, 1986); Perbal, A Practical Guide to Molecular Cloning (1984). Influenza A virus contains a genome consisting of 8 separate segments of negative sense RNA as shown in Table 1 below. (Steinhauer DA, Skehel JJ., 159890.doc -10- 201249996

Genetics of influenza viruses. Ann Rev Genet. 2002 36· 305-332). Circulating human strains are well known for their frequent accumulation of mutations year after year and causing epidemics. However, the segmentation of the genome allows for the exchange of intact genes between different strains. The genetic plasticity of influenza viruses is also likely to be related to vaccine design, pathogenicity, and the ability of new viruses to emerge from natural repositories and cause worldwide pandemics. Table 1. Epidemic's flu virus genome consists of 8 segments of s/s Μ 和 and # segment · size (10) peptide function 1 2341 PB2 transcriptase: cap binding 2 2341 PB1 transcriptase: elongation 3 2233 PA transcriptase: Protease (7) 4 1778 HA Hemagglutinin 5 1565 NP Nucleoprotein: RNA Binding 6 1413 NA Neuraminidase: Virus Release 7 1027 Ml Matrix Protein: Viral Component M2 Intrinsic Membrane Protein 8 890 NS1 Non-structural: Nuclear NS2 Non Structural: nuclear + cytoplasmic separation of total δ 190 viruses by cell culture 'A total of 193 neuraminidase (ΝΑ) and 197 hemagglutinin (ΗΑ) partial gene sequences were obtained from these viruses via capillary sequencing. Due to the most variable sequence of ΗΑ and ΝΑ1ΑΗ1, computer simulation analysis (pedigree analysis and BLAST) showed that all 386 (HA and NA sequences) can be divided into 20 clusters; in 454 Titanium Roche pyrosequencing equipment ( Pyrosequencing of the complete genome of a representative member of each cluster was performed in 454 Titanium Roche Pyrosequencing equipment, ROCHE). A total of 20 complete AH1N1 genomes were obtained, each consisting of 8 genes each AH1N1 159890.doc 201249996. The sequencing data revealed four viral isolates with three different mutations: (1) pAHIN1 virus with mutations in the NA (oseltamivir R), HA (hypervirulence) and stromal segments (virus 1); (ii) pAHIN1 virus, which has a mutation in the NA and stromal segments (virus 2); (iii) pAHIN1 virus, which has a mutation in the matrix (MA) segment (virus 3); and (iv) pAHIN1 virus, which is in the nerve There is a mutation in the aminase (NA) segment (virus 4). Changes in the hemagglutinin HA1 gene sequence at amino acid corresponding to position 222 (position 225 of the H3 number) (where aspartate (D) is changed to glycine (G)) is associated with severe clinical outcome . D222G mutations were recorded in the global isolate of HINlpdm in the gene pool at various time points between April and November 2009 (Chen GW, Shih SR (2009) Genomic signatures of influenza A pandemic (H1N1) 2009 virus. Emerg Infect Dis). The preferential combination of influenza virus with sialic acid-α2,3-galactose (α2,3 receptor) or sialic acid-α2,6-galactose (α2,6 receptor) determines its tropism. The α2,3 receptor and α2,6 receptors are dominant in lower respiratory tract cells and upper respiratory tract cells, respectively (Shinya Κ, Ebina Μ, Yamada S, Ono M, Kasai N, Kawaoka Y. Avian flu: influenza receptors in the human Airway. Nature. 2006; 440(7083):435-6) o The latest glycan microarray analysis showed that hemagglutinin (HA) D222G substitution may be caused by α2,6 receptor specificity to mixed α2,3/α2, 6 changes in receptor specificity, which enhances binding to the α2,3 receptor and increases disease severity. (Stevens J, Blixt Ο, Glaser L, Taubenberger JK, Palese 159890.doc 12 201249996 P, Paulson JC, et al., Glycan microarray analysis of the hemagglutinins from modern and pandemic influenza viruses reveals different receptor specificities. J Mol Biol. 2006 J 355 (5): 1 143-55). This position appears to affect receptor binding specificity, and the D/G difference between the two 1918 virus variants (the same as before) differs from the a2-6 linked sialic acid preference to the dual alpha2-3/α2·6 specificity. The change is related to the fact that it may make the virus more susceptible to infection in the airway deeper (where the cells expressing the α2-3 receptor are more abundant), thereby producing hypervirulence. However, a unique pedigree population of severe cases with D222G substitution has not been observed (Mak GC et al, Association of D222G substitution in hemagglutinin of 2009 pandemic influenza A (H1N1) with severe disease. Eurosurveillance, Vol. 15, No. 14, April 8th, 2010). All of the viruses found to have a change in position 222 in HA1 belong to the sub-branches of the pandemic H1N1 virus characterized by HA1 substitution S203T (characteristic of branch 7), which is also surrounded by an amino acid bag and the amine group The acid bag is exposed to the antigenic area unique to this supervirulent strain and the remaining AH1N1 strain. This sub-branch accounts for most of the pandemic virus in Europe and the rest of the northern hemisphere. For the southern hemisphere, this sub-branch is also widely distributed. Based on the currently available data shared with WHO, the prevalence of D222G replacement is less than 1.8% (52 detected in more than 2755 HA sequences). Of the 364 fatal cases analyzed to date, 26 of the cases (7.1%) had D222G substitution. There is limited clinical information about possible potential medical conditions in these cases. The pandemic (H1N1) 2009 virus with D222G substitution is similar to the A/California/7/2009 (H1N1) virus (WHO recommended vaccine virus) in 159890.doc •13·201249996. In the same virus A (H1N1) 2009, it was found that (i) hemagglutinin (HA) _ Gly222 mutation and (ii) neuraminidase (NA) Tyr 275 associated with resistance to oseltamivir Mutations (McKimm-Breschkin JL. Resistance of influenza viruses to neuraminidase inhibitors -a review. Antiviral Res 2000; 47:1-17; Collins PJ et al.

Structural basis for oseltamivir resistance of influenza viruses. Vaccine 27 (2009) 6317-6323). Since both mutations are associated with important phenotypes (supervirulence and oseltamivir resistance), and the antigenic regions are associated with their mutations, the vaccine candidates of the present invention can effectively recognize mutant strains and AH1N1 wild type II. By. Three D222G variant viruses contain H275Y substitutions associated with oseltamivir resistance in neuraminidase (NA). WHO Report. Preliminary review of D222G amino acid substitution in the hemagglutinin of pandemic influenza A (H1N1) 2009 viruses, December 28, 2009 (www.who.int/csr/resources/publications/swineflu/cpl65_ 2009_2812_review_d222g_amino_acid_substitution_in_ha_ hlnl_viruses.pdf) o The currently available pandemic vaccine also recognizes hypervirulence and oseltamivir resistant mutants, however, although the mutation is located in the receptor binding pocket of H1 hemagglutinin rather than the antigenic site, the infectious disease in Sweden The preliminary genetic characterization of the Swedish Institute for Infectious Disease Control and our laboratory showed that compared with the wild-type pandemic AH1N1 strain 159890.doc •14-201249996 than the 'mutant virus and the current pandemic The reactivity of the antisera of the vaccine is significantly reduced. Therefore, the current vaccines are not as effective in identifying new mutant strains that spread rapidly around the world as the vaccine candidates of the present invention. The mutation in the 3, end of the viral genome segment 7 (stromal segment) showed similarity to the epidemic • estrus section (60%). This mutation has previously been reported to increase the specific immune response in humans and it can also confer protection against influenza. However, cell-mediated immunity, primarily resistant to viral matrices and nuclear protein antigens, does not protect against infection, but it is the key to viral clearance and recovery from disease. 159890.doc •15· 201249996 (l ♦ 螺)跏钬枇¥-&-趔11 Whore? 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wlvoxDwlIlvllDDVuLU £ aIO.UXDOVOWlvovovvuwoVHOVVmioaIDVlDWDVJLLD DowoolvlllvuVVIVVVuvvoulllDVIWDVovuVDDOVVVaLLVDVuvooouDIVVVoVVOIXVVJXOaLSW IVWOovaLLLVWOVuVOOOLLWDOVOVODODlovvvvaLWVOVDOVDVoaLDuVXDVollDuvovooaLVVOV OHVOHOlDVWovsllDsllvovovuvlvlvvvvolDlulovoulloouvovvloaLVVOOHVVaLLDVOOVosl voXDU18ooI-LVVDolvvvvvDHolvsvaLv.ll3vHvuVODVouIlDlDIlDuVJ.LVDovo.LLLWVOVOaLLOVHv VDOVOOOVOOVOIDVaLLLDOOOVVXDVJLDVlvoBovovov; LLV8DvvlxL31vvHolovaLIOW80vvvoluv lovlvoWDWWOaLVDlvoDDOVuuvoaLLVODDHODOXXlDlDoDoovlwlvowwooovlDualvuIoolvo JJLODIDVOVDuvalaLVVVDDVOOlDUovolvuVOIDIllDoVIDODOullDVIDovlaLOHVDaLLWuvaLWoVIl uvuVDVODOVUuHWOOVOVWOLIDlvsHVVDalouHVVVUWVDDOlloIJalDVuoovuVVVDuHvlvslDuo OXIODDVDWDDDVOW90vvwllDDlowvvuHVOVODDOo3XDOOalvaLLDaxxv8801volooaLLVOaIV vuv0v3OUIV0D0VVD000lvHVDDOXLI: mLI01DVDVlsvaLLl: > OVVD10B0v: > w3uBVDaLWE- v3vv (2ε ON QIcyHS) -8-inch drive 11 € bee! wlovDV!DDVOVlvuvvuloIlIlvolDaLV8v9110IVDaLO 159890.doc •43- 201249996 The amino acid sequence of four virus isolates was derived from nucleotide sequence determination inferred. Virus 1 carries a mutation of interest in the HA, NA and MA proteins. Virus 2 carries a mutation of interest in the NA and MA proteins. Virus 3 carries a mutation of interest in the NA protein. Virus 4 carries a mutation of interest in the MA protein. The mutations of interest in the HA sequence were analyzed relative to the wild-type pandemic influenza Η1N1 sequence. All four isolates contained mutations P83S and 1321V 'The mutations appeared to be prevalent in branch 7 of the Α/Η1Ν1 pandemic strain. Virus 1 also has (1) branch 7 characteristic S203T mutation, and (ii) located in ΗΑ D222E mutation at position 222. At positions 222-223, virus 1 has an E-R amino acid rather than a wild-type D-Q. An exact match to the HA sequence of virus 1 was found in the influenza A virus (A/Novgorod/01/2009 (HlNl)) HA protein. 4- wild-type segment 1A] Influenza A / Ca Shu ifornia / 07/2009 sequence (FJ966974) MKAILWLLY TFATANADTL CIGYHANNST DTVDTVLEKN VTVTHSVNLL EDKHNGKLCK LRGVAPLHLG KCNIAGWILG NPECESLSTA SSWSYIVETg SSDNGTCVPG DFIDYEELRE QLSSVSSFER FEIFPKTSSW PNHDSNKGVT AACPHAGAKS FYKNLIWLVK KGNSYPKLSK SYINDKGKEV LVLWGIHHPS TSADQQSLYQ NADAYVFV exhaust SRYSKKFKPE IAIRPKVR® EGRMNYYWTL VEPGDKITFE ATGNLWPRY AFAMERNAGS GIIISDTPVH DCNTTCQTPK GAINTSLPFQ NIHPITIGKC PKYVKSTKLR LATGLR ^ PS IQSRGLFGAI AGFIEGGWTG MVDGWYGYHH QNEQGSGYAA DLKSTQNAID EITNKVNSVI EKMNTQFTAV GKEFNHLEKR IENLNKKVDD GFLDIWTYNA ELLVLLENER TLDYHDSNVK NLYEKVRSQL KNNAKEIGNG CFEFYHKCDN TCMESVKNGT YDYPKYSEEA KLNREEIDGV KLESTRIYQI LAIYSTVASS LVLVVSLGAI SFWMCSNGSL QCRICI (SEQ ID NO; 33) - virus 1 MKAILWLLY TFATANADTL CIGYHANNST DTVDTVLEKN VTVTHSVNLL EDKHNGKLCK 60 • 44 - 159890.doc 201249996

LRGVAPLHLG KCNIAGWILG NPECESLSTA SSWSYIVETS SSDNGTCYPG DFIDYEELRE 120 QLSSVSSFER FEIFPKTSSW PDHDSNKGVT AACPHAGAKS FYKNLIWLVK KGNSYPKLSK 180 SYINDKGKEV LVLWGIHHPS TSADQQSLYQ NADAYVFVGf SRYSKKFKPE IAIRPKVRgg 240 EGRMNYYWTL VEPGDKITFE ATGNLWPRY AFAMERNAGS GIIISDTPVH DCNTTCQTPK 300 GAINTSLPFQ NIHPITIGKC PKYVKSTKLR LATGLRNVPS IQSRGLFGAI AGFIEGGWTG 360 MVDGWYGYHH QNEQGSGYAA DLKSTQNAID EITNKVNSVI EKMNTQFTAV GKEFNHLEKR 420 IENLNKKVDD GFLDIWTYNA ELLVLLENER TLDYHDSNVK NLYEKVRSQL KNNAKEIGNG 480 CFEFYHKCDN TCMESVKNGT YDYPKYSEEA KLNREEIDGV KLESTRIYQI LAIYSTVASS 540 LVLWSLGAI SFWMCSNGSL · QCRICI (SEQ ID NO: 34) 566 virus 2 MKAILWLLY TFATANADTL CIGYHANNST DTVDTVLEKN VTVTHSVNLL EDKHNGKLCK LRGVAPLHLG KCNIAGWILG NPECESLSTA SSWSYIVET§ SSDNGTCYPG DFIDYEELRE QLSSVSSFER FEIFPKTSSW PNHDSNKGVT AACPHAGAKS FYKNLIWLVK KGNSYPKLSK SYINDKGKEV LVLWGIHHPS TSADQQSLYQ NADAYVFVi ^ SRYSKKFKPE IAIRPKV ah EGRMNYYWTL VEPGDKITFE ATGNLWPRY AFAMERNAGS GIIISDTPVH DCNTTCQTPK GAINTSLPFQ NIHPITIG KC PKYVKSTKLR LATGLRNyPS IQSRGLFGAI AGFIEGGWTG • 45· 159890.doc 201249996

MVDGWYGYHH QNEQGSGYAA DLKSTQNAID EITNKVNSVI EKMNTQFTAV GKEFNHLEKR IENLNKKVDD GFLDIWTYNA ELLVLLENER TLDYHDSNVK NLYEKVRSQL KNNAKEIGNG CFEFYHKCDN TCMESVKNGT YDVPKYSEEA KLNREEIDGV KLESTRIYQI LAIYSTVASS LVLWSLGAI SFWMCSNGSL QCRICI (SEQ ID Ϊ 35) virus 3 MKAILWLLY TFATANADTL CIGYHANNST DTVDTVLEKN VTVTHSVNLL EDKHNGKLCK LRGVAPLHLG KCNIAGWILG NPECESLSTA SSWSYIVETS SSDNGTCYPG DFIDYEELRE QLSSVSSFER FEIFPKTSSW PNHDSNKGVT AACPHAGAKS FYKNLIWLVK KGNSYPKLSK SYINDKGKEV LVLWGIHHPS TSADQQSLYQ NADAYVFVG for SRYSKKFKPE IAIRPKVi ^ EGRMNYYWTL VEPGDKITFE ATGNLWPRY AFAMERNAGS GIIISDTPVH DCNTTCQTPK GAINTSLPFQ NIHPITIGKC PKYVKSTKLR LATGLRN ^ PS IQSRGLFGAI AGFIEGGWTG MVDGWYGYHH QNEQGSGYAA DLKSTQNAID EITNKVNSVI EKMNTQFTAV GKEFNHLEKR IENLNKKVDD GFLDIWTYNA ELLVLLENER TLDYHDSNVK MLYEKVRSQL KNNAKEIGNG CFEFYHKCDN TCMESVKNGT YDYPKYSEEA KLNREEIDGV KLESTRIYQI LAIYSTVASS LVLWSLGAI SFWMCSNGSL QCRICI (SEQ ID NO: 36) virus 4. · MKAILWLLY TFATANADTL CIGYHANNST DTVDTVLEKN VTVTHSVNLL EDKHNGKLCK -46· 1598 90.doc 201249996 LRGVAPLHLG KCNIAGWILG NPECESLSTA SSWSYIVETS SSDNGTCYPG DFIDYEELRE - QLSSVSSFER FEIFPKTSSW PNHDSNKGVT AACPHAGAKS FYKNLIWLVK KGNSYPKLSK SYINDKGKEV LVLWGIHHPS TSADQQSLYQ NADAYVFVGg SRYSKKFKPE IAIRPKV La EGRMNYYWTL VEPGDKITFE ATGNLWPRY AFAMERNAGS GIIISDTPVH DCNTTCQTPK GAINTSLPFQ NIHPITIGKC PKYVKSTKLR LATGL_PS IQSRGLFGAI AGFIEGGWTG MVDGWYGYHH QNEQGSGYAA DLKSTQNAID EITNKVNSVI EKMNTQFTAV GKEFNHLEKR IENLNKKVDD GFLDIWTYNA ELLVLLENER TLDYHDSNVK NLYEKVRSQL KNNAKEIGNG CFEFYHKCDN TCMESVKNGT YDYPKYSEEA KLNREEIDGV KLESTRIYQI LAIYSTVASS LVLWSLGAI SFWMCSNGSL QCRICI (SEQ ID NO: 37) Virus 1, 2 and 4 are carried at the I106V, D248N, Y275H residues of the NA segment (6) relative to the wild type A/California/07/2009 sequence. mutation. The Y275H mutation is associated with oseltamivir resistance. The exact match of the NA sequences found in the three isolates was also found in other isolates. Current wild-type vaccines are less effective against virus 1 containing both HA and NA mutations at these specific sites. (Puzelli et al., Transmission of Hemagglutinin D222G mutant strain of pandemic (H1N1) 2009 virus. Emerging Infectious Diseases 16(5): 863-865 (2010)). Vaccine generated by a virus (eg, virus 1) containing both hypertoxicity (D222G/E in the HA region) and oseltamivir resistance (Y275H in the NA region), a wild-type H1N1 pandemic vaccine It is inefficient and provides protection against both hypervirulence and oseltamivir-resistant viruses. -47· 159890.doc 201249996

Zone 6-NAA / California / 07/2009 sequence MNPNQKIITI GSVCMTIGMA NLILQIGNII SIWISHSIQL GNQNQIETCN QSVITYENNT WVNQTYVNIS NXNFAAGQSV VSVKLAGNSS LCPVSGWAIY SKDNSgRIGS KGDVFVIREP FISCSPLECR TFFLTQGALL NDKHSNGTIK DRSPYRTLMS CPIGEVPSPY NSRFESVAWS ASACHDGINW LTIGISGPDN GAVAVLKYNG IITDTIKSWR NNILRTQESE CACVNGSCFT VMTDGPSgGQ ASYKIFRIEK GKIVKSVEMN APNYgYEECS CYPDSSEITC VCRDNWHGSN RPWVSFNQNL EYQIGYICSG IFGDNPRPND KTGSCGPVSS NGANGVKGFS FKYGNGVWIG RTKSISSRNG FEMIWDPNGW TGTDNNFSIK QDIVGINEWS GYSGSFVQHP ELTGLDCIRP CFWVELIRGR PKENTIWTSG SSISFCGVNS DTVGWSWPDG AELPFTIDK (SEQ ID NO: 38) virus 1 MNPNQKIITI GSVCMTIGMA NLILQIGNII SIWISHSIQL GNQNQIETCN QSVITYENNT WVNQTYVNIS NTNFAAGQSV VSVKLAGNSS LCPVSGWAIY SKDNS | RIGS KGDVFVIREP FISCSPLECR TFFLTQGALL NDKHSNGTIK 050 100 150 200 250 300 350 400 450 469 50 100 150 159890. Doc 48- 201249996

DRSPYRTLMS CPIGEVPSPY NSRFESVAWS ASACHDGINW LTIGISGPDN 200 GAVAVLKYNG IITDTIKSWR NNILRTQESE CACVNGSCFT VMTDGPS0GQ 250 ASYKIFRIEK GKIVKSVEMN APNYg ^ EECS CYPDSSEITC VCRDNWHGSN 300 RPWVSFNQNL EYQIGYICSG IFGDNPRPND KTGSCGPVSS NGANGVKGFS 350 FKYGNGVWIG RTKSISSRNG FEMIWDPNGW TGTDNNPSIK QDIVGINEWS 400 GYSGSFVQHP ELTGLDCIRP CFWVELIRGR PKENTIWTSG SSISFCGVNS 450 DTVGWSWPDG AELPFTIDK (SEQ ID NO: 39) 469 virus 2 MNPNQKIITI GSVCMTIGMA NLILQIGNII SIWISHSIQL GNQNQIETCN QSVITYENNT WVNQTYVNIS NTNFAAGQSV VSVKLAGNSS LCPVSGWAIY SKDNSgRIGS KGDVFVIREP FISCSPLECR TFFLTQGALL NDKHSNGTIK DRSPYRTLMS CPIGEVPSPY NSRFESVAWS ASACHDGINW LTIGISGPDN GAVAVLKYNG IITDTIKSWR NNILRTQESE CACVNGSCFT VMTDGPSfGQ ASYKIFRIEK GKIVKSVEMN apnyJyeecs CYPDSSEITC VCRDNWHGSN RPWVSFNQNL EYQIGYICSG IFGDNPRPND KTGSCGPVSS NGANGVKGFS FKYGNGVWIG RTKSISSRNG FEMIWDPNGW TGTDNNFSIK QDIVGINEWS GYSGSFVQHP ELTGLDCIRP CFWVELIRGR PKENTIWTSG SSISFCGVNS • 49- 159890. Doc 201249996 DTVGWSWPDG AELPFTIDK (SEQ ID NO: 40) Virus 3

MNPNQKIITI GSVCMTIGMA NLMLQIGNII SIWISHSIQL · GNQNQIETCN QSVITYENNT WVNQTYVNIS NTNFAAGQSV VSVKLAGNSS LCPVSGWAIY SKDN ^ RIGS KGDVFVIREP FISCSPLECR TFFLTQGALL NDKHSNGTIK DRSPYRTLMS CPIGEVPSPY NSRFESVAWS ASACHDGINW LTIGISGPDN GAVAVLKYNG IITDTIKSWR NNILRTQESE CACVNGSCFT VMTDGPSgEQ ASYKIFRIEK GKIVKSVEMN APNYgYEECS CYPDSSEITC VCRDNWHGSN

RPWVSFNQNL EYQIGYICSG IFGDNPRPND KTGSCGPVSS NGANGVKGFS FKYGNGVWIG RTKSISSRNG FEMIWDPNGW TGTDNNFSIK QDIVGINEWS GYSGSFVQHP ELTGLDCIRP CFWVELIRGR PKENTIWTSG SSISFCGVNS DTVGWSWPDG AELPFTIDK (SEQ ID NO: 41) Virus 4

MNPNQKIITI GSVCMTIGMA NLILQIGNII SIWISHSIQL GNQNQIETCN QSVITYENNT WVNQTYVNIS NTNFAAGQSV VSVKLAGNSS LCPVSGWAIY SKDNS | RIGS KGDVFVIREP FISCSPLECR TFFLTQGALL NDKHSNGTIK DRSPYRTLMS CPIGEVPSPY NSRFESVAWS ASACHDGINW LTIGISGPDN 159890.doc 50- 201249996 GAVAVLKYNG IITDTIKSWR NNILRTQESE CACVNGSCFT VMTDGPS | GQ ~

ASYKIFRIEK GKIVKSVEMN APNYgYEECS CYPDSSEITC VCRDNWHGSN

RPWVSFNQNL EYQIGYICSG IFGDNPRPND KXGSCGPVSS NGANGVKGFS

FKYGNGVWIG RTKSISSRNG FEMIWDPNGW TGTDNNFSIK QDIVGINEWS

GYSGSFVQHP ELXGLDCIRP CFWVELIRGR PKENTIWTSG SSISFCGVNS DIVGWSWPDG AELPFTIDK (SEQ ID NO: 42) The viruses 1, 2 and 3 contained the mutation A198P in the M1 region (Segment 7) relative to the wild type A/California/07/2009 sequence. Furthermore, the three isolates lack residues corresponding to D34 in the M2 region. This mutation has been reported to increase cell-mediated immunity (but does not protect against infection). The mutated region has 60% sequence similarity to the influenza B virus MA region and the vaccine against the virus isolate carrying the mutation produces immunity against influenza B. In addition, M1 mutations can be used as therapeutic targets for viral clearance and recovery. -51- 159890.doc 201249996

Section 7-MAA / Calif〇rnia / 07/2009 sequence Ml MSLLTEVETY VLSIIPSGPL KAEIAQRLES VFAGKNTDLE ALMEWLKTRP ILSPLTKGIL GFVFTLTVPS ERGLQRRRFV QNALNGNGDP NNMDRAVKLY KKLKREITFH GAKEVSLSYS TGALASCMGL IYNRMGTVTT EAAFGLVCAT CEQIADSQHR SHRQMATTTN PLIRHENRMV LASTTAKAME QMAGSSEi ^ A EAMEVANQTR QMVHAMRTIG THPSSSAGLK DDLLENLQAY QKRMGVQMQR FK (SEQ ID NO: 43) M2 MSLLTEVETP TRSEWECRCS DSSgPLVIAA NIIGILHLIL WITDRLFFKC IYRRFKYGLK RGPSTEGVPE SMREEYQQEQ QSAVDVDDGH FVNIELE (SEQ ID NO: 44) virus 1 Ml MSLLTEVETY VLSIIPSGPL KAEIAQRLES VFAGKNTDLE ALMEWLKTRP 050 100 150 200 250 252 50 97 050 159890.doc 52 · 201249996 ILSPLTKGIL GFVFTLTVPS ERGLQRRRFV QNALNGNGDP NNMDI ^ AVKLY loo - KKLKREXTFH GAKEVSLSYS TGALASCMGL IYNRMGTVTT EAAFGLVCAT 150 CEQIADSQHR SHRQMATTTN PLIRHENRMV LASTTAKAME QMAGSSE ^ A 200 EAMEVANQTR QMVHAMRTIG THPSSSAGLK DDLLENLQAY QKRMGVQMQR 250 FK {SEQ ID NO: 45) 252 M2 MSLLTEVETP TRSEWECRCS DSS | PLVIAA NIIGILHLIL WITDRLFFKC 50 IYRRFKYGLK RGPSTEGVPE SMREEYQQEQ QSAVDVDDGH FVNIELE 96 (SEQ ID NO: 46 ) Virus 2 Ml MSLLTEVETY VLSIIPSGPL KAEIAQRLES VFAGKNTDLE ALMEWLKTRP 050 ILSPLTKGIL GFVFTLTVPS ERGLQRRRFV QNALNGNGDP NNMDHAVKLY 100 KKLKREITFH GAKEVSLSYS TGALASCMGL IYNRMGTVTT EAAFGLVCAT 150 CEQIADSQHR SHRQMATTTN PLIRHENRMV LASTTAKAME QMAGSSEC ^ V 200 EAMEVANQTR QMVHAMRTIG THPSSSAGLK DDLLENLQAY QKRMGVQMQR 250 FK (SEQ ID NO: 47) 252 -53- 159890.doc 201249996 M2 MSLLTEVETP TRSEWECRCS DSS | PLVIAA NIIGILHLIL WITDRLFFKC IYRRFKYGLK RGPSTEGVPE SMREEYQQEQ QSAVDVDDGH FVNIELE (S £ Q ID NO: 48) virus 3Ml MSLLTEVEXY VLSIIPSGPL KAEIAQRLES VFAGKNTDLE ALMEWLKTRP ILSPLTKGIL GFVFTLTVPS ERGLQRRRFV QNALNGNGDP NNMDRAVKLY KKLKREITFH GAKEVSLSYS TGALASCMGL IYNRMGTVTT EAAFGLVCAT CEQIADSQHR SHRQMATTTN PLIRHENRMV LASTTAKAME QMAGSSEQ0A EAMEVANQTR QMVHAMRTIG THPSSSAGLK DDLLENLQAY QKRMGVQMQR FK (SEQ ID NO: 49) M2 MSLLTEVETP TRSEWECRCS DSS|PLVIAA NIIGILHLIL WITDRLFFKC IYRRFKYGLK RGPSTEGVPE SMREEYQQEQ QSAVDVDDGH FVNIELE (SEQ ID NO: 50) Virus 4 50 96 050 100 150 200 250 252 50 96 159890.doc 54· 201249996

Ml MSLLTEVETY VLSIIPSGPL KAEIAQRLES VFAGKNTDLE ALMEWLKTRP 050 ILSPLTKGIL GFVFTLTVPS ERGLQRRRFV QNALNGNGDP NNMDRAVKLY 100 KKLKREITFH GAKEVSLSYS TGALASCMGL IYNRMGTVTT EAAFGLVCAT 150 CEQXADSQHR SHRQMATTTN PLIRHENRMV LASTTAKAME QMAGSSEC ^ A 200 EAMEVANQTR QMVHAMRTIG THPSSSAGLK DDLLENLQAY QKRMGVQMQR 250 FK (SEQ ID NO: 51) 252 M2 MSLLTEVETP TRSEWECRCS DSSgPLVIAA NIIGILHLIL WITDRLFFKC 50 IYRRFKYGLK RGPSTEGVPE SMREEVQQEQ QSAVDVDDGH rVNIELE 97 (SEQ ID NO: 52) The method of the invention uses the influenza virus antigens disclosed herein to immunize against influenza virus infection. The particular virus from which the antigen is derived may be the same as or different from the particular virus against which the protection is provided, as it is known to exist between the different isolates within the influenza virus, particularly the same virus subtype. Currently used influenza vaccines are designated as whole virus (wv) vaccines or subviral particles (sv) (also known as "cleavage" or "purified surface antigens"). The WV vaccine contains a complete inactivated virus, and the SV vaccine contains a purified virus that has been split by a detergent which dissolves the lipid-containing envelope membrane and then chemically inactivates the residual virus. Anti-influenza attenuated virus vaccines are also under development. A discussion of the preparation of conventional vaccines can be found in Wright, P. F. and Webster, R. G., FIELDS VIROLOGY, 4th Edition (Knipe, D. Μ. et al., ed.), 1464-65 (2001). If the vaccine comprises more than one influenza strain, the different strains are usually grown separately and mixed after the virus has been harvested and the antigen has been prepared. Alternatively, different segments of different isolates of influenza virus can be combined to produce a pluripotent vaccine. The influenza virus used in the methods of the invention may be a redistributed strain and/or may be obtained by reverse genetics techniques. The virus can be an attenuated virus. The virus can be a temperature sensitive virus. The virus can be a cold-blooded virus. Redistribution strains including 'self-pathogenic strains of ha and/or NA virus segments and from the remaining 6 or 7 segments of non-pathogenic strains can be used. The influenza virus antigen used in the immunogenic composition of the present invention may be in the form of a live virus or preferably an inactivated virus. Virus inactivation usually involves treatment with chemicals such as formalin or beta-propiolactone. If a non-activated virus is used, the antigen can be a whole virus, a split virus or a virus. The split virus is obtained by using a detergent (e.g., diethyl ether, polysorbate 80, deoxycholate, tris-butyl butyl phosphate, Trit〇n χ_1〇〇,

Triton® 101, cetyltrimethylammonium bromide, etc.) are treated to treat virions to produce a subviral particle preparation. The subunit vaccine comprises one or both of the influenza surface antigen hemagglutinin and neuraminidase. Epidemic antigens can also be presented in virion form. If the antigen is produced from a prevalent V-virus (i.e., not in a recombinant or synthetic system that is not involved in the growth of influenza virus), the virus can grow on the egg or in cell culture. Growth in embryonic eggs without specific pathogens allows the influenza virus to grow for the traditional route of vaccine production, and the cell culture system updates the development method. If cell culture is used, the influenza virus vaccine will usually grow on mammalian cells (e.g., MDCK cells, VERO cells, or pER.c6 cells). These cell lines can be used anywhere, for example, from the American Type Cell Culture (ATCC) collection or from the Coriell Cell Repositories. For example, the ATCC supplies a variety of different VERO cells&apos; with catalog numbers CCL-81, CCL-81.2, CRL-1586, and CRL-15 87 and supplies MDCK cells with catalog number CCL-34. It is also possible to grow on avian cell lines (including cell lines derived from hens, such as chicken embryo fibroblasts (CEF)). The immunogenic and pharmaceutical compositions of the invention are suitable for administration to a patient. This can be achieved by a variety of means including, but not limited to, intradermal injection; transdermal administration; and topical administration. Such means may be used in conjunction with, for example, skin emery or by skin abrasion resulting from the use of microabrasives. The immunogenic and pharmaceutical compositions of the invention are preferably in the form of a vaccine. Compositions of the invention may include an adjuvant. Adjuvants used in influenza vaccines include aluminum salts, chitosan, CpG deoxynucleotides (eg CpG 7909), oil-in-water emulsions (eg MF59), water-in-oil-in-water emulsions, Escherichia coli CE.co/z·) heat-labile toxin and its detoxifying mutant, monophosphorus a lipid and its 3-0-deuterated derivative, pertussis toxin mutant, cell wall quinone dipeptide, etc. . Hemagglutinin (HA) does not activate the primary immunogen in influenza vaccine, and the vaccine dose is normalized to the HA content and is usually measured by one-way radiation immunodiffusion (SRID) analysis. Intramuscular injection vaccines usually contain about 15 HA/strain, but lower doses (for example in children, or in a pandemic situation) can also be used and fractionated doses have been used, such as 1/2 (ie 7 5 gg) HA/strain), 1/4 and 1/8 dose. The compositions of the present invention will typically comprise between (M and 8 HA, preferably about 7, for example, about 5, about 3, about 2.5, about 2, about 2 per POP 159890.doc -57 - 201249996). K5, about i, about 75, about 5:, 〇.4, about 0.2, etc. The intramuscular injection vaccine usually has a volume of 〇5 ml. The composition may include a preservative such as thiomersal* 2_phenoxyethanol. However, the mouthpiece should preferably be substantially free (ie less than! The heart is called a material containing, for example, no thiomersal. A vaccine containing no mercury is better. If the influenza virus is already in the cell Growing on culture, the compositions of the invention preferably contain less than 100 pg of residual host cell DNA per dose but may be present in trace amounts of host cell DNA «can be used during vaccine preparation using standard purification procedures such as chromatography to remove contamination DNA. Removal of residual host cell DNA can be enhanced by nuclease treatment (for example, by using Benzonase 8 DNase). A vaccine containing &lt;100 pg host cell DNA/1 血 called hemagglutinin is preferred, containing &lt;1 〇〇pg host cell DNA/〇25 ml volume of vaccine is also compared A vaccine containing &lt;100 pg of host cell DNA/5 〇 hemagglutinin is preferred. A vaccine containing &lt;100 pg of host cell DNA/ml volume is also preferred. The vaccine of the present invention can be delivered according to a single dose immunization schedule. Alternatively, it can be delivered as a priming element of a prime-boost regimen (meaning a second injection of similar antigenicity within weeks or months after the first immunization). Testing the immunogen of the influenza vaccine The method of sex is well known in the art. One method involves the following procedures: (a) taking a 10 ml venous blood sample from the patient (usually from the arm) for baseline titration of circulating anti-ha antibodies immediately prior to vaccination; (b) The patient immediately received a dose of 1 vaccine. If the vaccine was administered 159890.doc -58- 201249996 and the arm should be given the contralateral arm of the blood collection arm; (C) 10 ml of blood sample should be taken from the patient about 3 weeks after vaccination. Separate serum from blood samples and store (if necessary) at -20. (.. Analyze serum-resistant anti-hemagglutinin antibodies by hemagglutination inhibition (HI) or one-way radiation hemolysis (SRH). Public benchmark laboratory Sexual and negative sera and reference preparations. Antibody titration was performed in duplicate 'and titration of pre-vaccination sera and vaccination. The titer assigned to each sample is the geometric mean of two independent determinations (but out of The calculation objective 'any HI result of <1〇 (= undetectable) under standard conditions is expressed as 5 and any negative srh results are expressed as 4 mm2) 〇 In the HI test, the seroconversion rate corresponds to 24 免疫 immunity Significant (eg, at least 4 fold) increase in pre-immunization post-immunization titer and antibody titer. In the SRH test, the clear conversion rate corresponds to the post-vaccination area of 225 MM2 and the area relative to at least 50% of the area prior to vaccination. EXAMPLES It is not intended to limit the scope of the invention, and the examples, apparatus, methods, and related results of the embodiments of the invention are set forth below. Note that for the convenience of the reader, the title or subtitle may be used in the examples, which should in no way limit the scope of the invention. In addition, certain theories are set forth and disclosed herein; however, the invention is in no way intended to limit the scope of the invention, and the invention is practiced in accordance with the invention, that is, τ, regardless of (4) specific theory or scheme of action. Example 1: Cell line VERO cell line A well-studied cell line, first isolated from African green monkey kidney (Peace monkey (Ce (10) state), and has been widely used in micro 159890.doc -59- 201249996 Biology , cell biology and virology research. It has been approved for use in the production of viral vaccines (rotavirus and polivirus) in the United States, which have also been tested and in some cases approved for use against rabies (Rabies), Rio Vaccine production of viruses (reovirus) and Japanese encephalitis. Since this cell line is susceptible to co-infection and it can grow on a commercial scale in a fermentor or bioreactor (a desirable feature of vaccine production), this cell line was chosen. The VERO cell line used was purchased from ATCC and maintained at -70 °C with dimethyl sulfoxide (DMSO). Thawing cells according to CORNING instructions Example 2: Sterile conditions All facilities in BSLIII have been carefully handled and monitored to avoid contamination. The room, material, reagent and equipment sterility tests were performed regularly. The cell culture area was regularly cleaned and disinfected every month, and several rounds of disinfection were performed every 24, 48, and 72 hours using Benzai, 70% ethanol, and 5% gas. The sterility test was performed at the end of the sterilization and all reagents were tested for contamination by culturing aliquots in the thioglycollate liquid medium. Example 3: Cell proliferation

Vero cell proliferation is carried out under suitable physical and chemical conditions (e.g., in polystyrene cell culture flasks (25 cm2 '50 cm2 and 75 cm2). The flasks were also subjected to gamma irradiation to reduce hydrophobicity. These containers allow for efficient cell adhesion and thus allow for the formation of a single layer. To promote cell proliferation, growth medium Μ199 or DMEM (Dulbecco's Modified Medium) and supplemented with 10% fetal bovine serum (BFS) 159890.doc * 60 - 201249996 (SIGMA-Aldrich); by using Phenol Red to monitor the maintenance of physiological pH. An anti-microbial mixture (penicillin, streptomycin, and amphotericin B) was added, and anti-PPLO was also added to inhibit fungal contamination. Cell cultures were incubated at 35 C / 5% C 〇 2 / 5% humidity. Cell quality and growth are monitored by phase contrast microscopy, which allows them to discard cultures that present bacterial or fungal contamination. Cell subculture was performed on flasks that were fully confluent as follows: 1. Remove and discard the medium. 2. Rinse the cell layer briefly with 0.25% (w/v) chymotrypsin-0.53 mM EDTA solution to remove traces of all serum containing trypsin inhibitor. 3. Add 2. 〇 mi to 3·〇 ml trypsin _ 〇 八 八 solution to the flask and observe the cells under a flip-chip microscope until the cell layer is dispersed (usually within 5 to 5 minutes). 4. Add 6.0 ml to 8. 〇 mi complete growth medium and aspirate the cells by slow pipetting. 5. Add an appropriate aliquot of the cell suspension to the new culture vessel. 6. Incubate the culture at 37 °C. When 80% confluence was observed, cells were seeded with ixi〇6 p AHN1 virus. This degree of confluence is best suited for observing the cytopathic effect caused by the influenza virus Α/Η1Νι. Example 4: Virus isolation Virus isolation was performed in a 25 cm2 flask inoculated with VERO cells (80% confluence). In the virus isolation, the growth medium was removed and the cells were subjected to three times 159890.doc -61 - 201249996 centrifugation and resuspension To discard cell debris. A 5 〇 μΐ swab sample clinically diagnosed with influenza AH1N1 and confirmed by real-time PCR was inoculated in the cell culture and incubated for 30 minutes to allow the virus to adhere to the cell surface. Thereafter, BFS 1% was supplemented to Μ199 and cultured at 4 CTC/40 min. The presence of cytopathic effects in the inoculated culture was monitored by phase contrast microscopy. At the same time, the viral load was determined by real-time PCR. Upon reaching ιχ1〇6, the culture was frozen at -80 °C (DMSO 10%) and stored for further study. Example 5: Confirmation and quantification of virus infection by instant phlegm (11). Viral RNA was extracted from VERO cell culture by using a commercially available kit (ROCHE) as previously described by spectrophotometry Method for quantification of RNA using the NanoDrop device. The CDC protocol for AH1N1 detection was performed by real-time PCR in a Light Cycler 480 Temperature Cycler (ROCHE), which includes standards for the quantification of influenza (selected in The β1 viral load assay for the H1 gene in the plasmid is based on the calculation of the dot-point format for each sample. Figure 1 shows an example of viral load quantification. Example 6: Sample selection from 3760 pAHINl A total of 193 sequencing samples for positive samples were based on clinical and epidemiological data ^ clinical selection criteria: infection severity, drug resistance, and atypical clinical environment. The epidemiological criteria were based on location, gender, and age. The selected samples were collected during the four outbreak peaks reported from March to November 2009. The samples were resuspended in 2.5 mL of virus delivery culture 159890.doc 62· 201249996 Nutrients 'Distributed 500 μΐ Samples were stored at _7 〇t. Example 7: Nucleic acid extraction Using the MagNA Pure LC Total Nucleic Acid Separation Kit (Roche) on the MagNA Pure LC instrument (R0Che) by external lysis according to the manufacturer's instructions (External) The Lysis) protocol separates the total nucleic acid from the clinical sample and resuspends it in deionized water (100 μ). Briefly: a 5 〇〇 sample is placed in a MagnaPure nucleic acid column and a volume of lysis buffer is added, after which, All parameters were set and separation started. The extracted RNA was placed at -70 ° C for further analysis. Example 8: Influenza p/AH1N1 real-time PCR detection was performed by two methods to perform pandemic influenza AH1N1/09 Detection: Instant-on-type influenza A/H1N1 test group (R〇che), using the ready-to-use RNA virus master kit (Roche). Heat was performed on a Light Cycler 2.0 instrument (Roche) under the following conditions: Cycle: 3〇min at 5〇°c; 15 min at 95°C; 45 cycles of 15 s at 94°C; and 30 s at 60°C 〇 The second method is used for epidemic A real-time RT-PCR CDC protocol for cold A (H1N1) using the primers described in the above protocol And probe and one-step RT-PCR Invitrogen SuperScriptTM III Platinum® - a step-by-step quantitative set. Thermal cycling was performed on a Light Cycler 2.0 instrument (R0Che) under 50 °C at 50 °C; at 95 °C 2 min; 45 cycles of 15 s at 95 ° C; and 30 s at 55 ° C » All reactions in both methods are based on the manufacturer's instructions for 5 μί total in 20 μί total reaction volume RNA is implemented. In both assays, the 159890.doc •63·201249996 method includes an internal control (human myostatin gene for the Roche method and RnaseP for the CDC protocol). Each sample was tested and the following probes were used separately for immediate PCR: influenza A (InfA), universal pig (SwFlu A), H1 pig (SwHl), and an internal control for human nucleic acid RNaseP (RP). Positive and negative controls were introduced. The reaction mixture contained 0.8 μM per primer, 0.2 μM per probe, IX PCR buffer, and 0.5 U of Superscript®/Platinum Taq Mix (Invitrogen). An example of a sample configuration is shown in Figure 2. The amplification conditions were as follows: Reverse transcription - 50 ° C / 30 min Not activated - 15 ° C / 2 min Extended by 95 ° C / 15 cycles and 55 ° C / 30 cycles. Example 9: Data Analysis The results were analyzed as follows according to the WHO criteria: 1 There should be no fluorescence in the negative control. 2. - The experimental sample should have the fluorescence of the RP probe. 3. - Human nucleic acid controls should not emit fluorescence from InfA, SwFluA and SwHl probes. 4. - Positive control (A/H1N1 nucleic acid) should emit fluorescence from all primers and probes. 5. - If all criteria are met, the sample should be considered positive. Figures 3A and 3B show the graphical window of the Light Cycler 2.0 Instant Temperature Cycler interface, which displays the fluorescence curve. Figure 3A) Clinical samples. Figure 3B) 159890.doc -64- 201249996 RNAseP (RP) assay. Example 10: Sequencing A total of 950 viruses were isolated by cell culture, and 193 neuraminidase (NA) and 197 hemagglutinin (HA) fractions were obtained therefrom via capillary sequencing in an aBI 3 100 sequencing machine (Applied Biosystems). gene sequence. Due to the most variable sequence in HA and NA lines AH1N1, computer simulation analysis (pedigree analysis and BLAST) showed that all 386 (HA and NA sequences) can be divided into 20 clusters; in 454 Tiro-Roche pyrosequencing equipment ( 454 Titanium

Pyrophosphate sequencing of the complete genome of a representative member of each family in Roche Pyrosequencing equipment, R〇CHE) Example 11: Proliferation of influenza virus in chicken embryos Several attempts have been made in the industry to make influenza vaccines Candidates are recommended to proliferate in VERO cells. However, these attempts initially failed due to the extremely low amount of recovered virus even after three passages. Consider this event's one or two subcultures in SPF (no specific pathogen) chicken embryos. The candidates were inoculated into the amniotic fluid with 0.1 mL per candidate. It also includes 40 other viruses previously isolated during the influenza pandemic, and each virus is vaccinated with two eggs. After three days at 35 ° C, amniotic fluid was harvested from each infected embryo. 'The collected liquid was centrifuged at 3 〇〇〇 X g to remove cell debris, and the supernatant was analyzed by qRT-pCR and stored at -80 . (: Next. All procedures are implemented in BSL 111 to comply with the required standards to avoid biosafety risks. After the passage, 'the virus load in three embryos is extremely high' is one of the virus candidates. Another ten isolates of 159890.doc-65-201249996 strain were tested by RT-PCR test, but showed low viral load. Therefore, they were re-inoculated in another SPF chicken embryo, but in this case inoculated to the allantoic sac In the solution, the virus was harvested by collecting only the allantoic fluid after 72 hours of inoculation under the same conditions as above. The smaller volume was analyzed by RT-PCR, and the remainder was stored at -80 ° C. The results showed that Six of the 10 virus isolates were able to replicate with high efficiency, resulting in a high viral load. Proof of the sequence from the Karolinska Institute. Before the chicken embryos were propagated, the viral strain cDNA was sent to Karolinska for research by using 454 pyrophosphate. The sequencer system ROCHE is used to verify the sequence. The bioinformatics analysis confirms the sequence previously determined in our laboratory. The master seed identifies the virus candidate for the influenza vaccine as H ZFLUJAL. Adaptation of influenza virus to growth on VERO cell lines Inoculates the embryonic harvested virus candidate (HZFLUJAL) and two other viruses on the VERO cell line. To achieve adaptation, several subcultures were performed and Culture supernatants were collected after 7 to 10 days of infection. Each supernatant was used to infect new VERO cell passages. The viral load for each passage was quantified. Figure 4A shows the same virus in embryos and individual passages. The difference between the viral loads in the culture. It is shown in the figure that up to four subcultures, the virus achieves adaptation to this tissue cell line. Influenza virus adapts to growth on MDCK cell lines not only VERO cells It was assessed as a matrix for influenza virus, and MDCK cell lines were also used to proliferate the virus. The same procedure was used to achieve adaptation to VER◦ cells in MDCK cells. Preliminary results showed 159890.doc •66· 201249996 Better adaptation to this cell line is due to the fact that it takes only three passages to obtain a viral load greater than that of the chicken embryo (Fig. 4B).

The virus was propagated to obtain the main species HZFLUJAL. For the proliferation of the virus which had been previously adapted to grow on the VERO cell line, the fourth subculture of the VERO cells of one cryotube was thawed and infected with VER0 on a j 75 cm2 flask. cell. After 33 days in 33 «&gt;c in the c〇2 incubator, 120 mL of the main species was obtained, which showed a viral load greater than 1〇7 virus particles/mL (approximately 1〇7·7ρ obtained 12〇) The cold-tube with i mL main species. The quality control of the main species is performed on the main species for sterility test and hemagglutination test. For the first test, 'the main species of 4 cryotubes are thawed, and two cryotubes are used to inoculate. Two flasks containing 100 mL of liquid thioglycollate medium NIH, and the other two cryotubes were used to inoculate the antibiotic-free DMEM medium. The two media were incubated for 8 to 15 days at 35 C. No observation was observed in the medium after the incubation period. To growth and change, it is considered as sterile (bacteria, fungi and protozoa). The blood coagulation test is performed to determine the titer of the lectin in the main species. In this analysis, the 96-well U-shaped Serial dilutions of 1:2 to 1:1024 of the main species were prepared in PBS (pH=7.4) in microplates. The same volume of 0.5% (v/v) human erythrocyte suspension stored in pbs was added to In each dilution, the microplate was placed at 4. (: 2 hours under cultivation) and the results showed blood. The agglutination titer is 1:8 » Example 12: Preclinical pilot test using the HZFLUJAL vaccine (Pre_ I59890.doc -67· 201249996 preclinical trial) The inactivation of the virus for preclinical prodromal testing is the implementation of preclinical guidance. The test was carried out to make 1 cold; the main species of the East Tube was untwisted, and the VER〇 culture was infected with it at 33 (after 7 days in the (:02 incubator) on the 8 () 2 flask. At 35. The final concentration was . Fine (v/v) fumarine did not activate the culture for 18 h. The fumarin was cleared by dialysis against sputum (10) four times in 48 h. After that, the hemagglutinin titer was performed. Quantification, which again adjusted the inactivated virus suspension to 1:6 virions/25 pL at 1:8 ° and used as a vaccine in the following groups. Preclinical pre-test animal models in preclinical pilot studies Female balb/c mice were used for 7 to 8 weeks. Each group was placed on a rack with a cage HePa filter cover. All animals should be treated to avoid unnecessary pain. At the completion of the protocol, all mice were euthanized. Analysis of seroconversion rates in preclinical pre-tests in 9 groups (5 small per group) Perform seroconversion analysis. Wei 1 vaccine with 10 viral particles + transfer factor + sublingual administration group 2 vaccine with 1 〇 6 viral particles + transfer factor + intramuscular administration group 3 with 1 〇 Vaccine granule vaccine + sublingual administration group 4 vaccine with 1 病毒 virus granule + intramuscular administration group 5 commercial vaccine + sublingual administration group 6 commercial vaccine + intramuscular injection group 7 fetal cattle Serum DMEM + sublingual administration of group 8 fetal fetal bovine serum DMEM + intramuscular administration 159890.doc -68 - 201249996 Group 9 untreated preclinical material test analysis in 5 groups (3 mice per group) Dosage response analysis was performed. Group A has 1 〇 7 virus granules vaccine + transfer factor + sublingual administration, Wei B has 1 病毒 virus granule vaccine + transfer factor + sublingual administration, and C has 1 病毒 virus granule vaccine + transfer factor + sublingual administration group D with 1 〇 3 viral particles of vaccine + transfer factor + sublingual administration group E with 1 病毒 virus granule vaccine + transfer factor + sublingual administration and self-immunization program, All mice were maintained for 4 days in an animal experimental room with biosafety requirements for performing immunoassays. After the time of adaptation, clinical data such as body weight and appearance were recorded, and no anticoagulant blood samples were taken from each mouse prior to immunization. Thereafter, each group was vaccinated by using separate administration routes. Four blood samples were obtained from each mouse on days 7, 14, 21 and 28 after immunization. Serum was obtained by centrifugation of blood samples and used to detect igG2 antibodies against influenza A H1N1 virus. In addition, secretory samples were collected from mice for detection of secreted immunoglobulin A (IgA). Example 13: Detection of anti-influenza p/AHINl antibodies produced by CIATEJ/OPKO/vaccines The actual v-type vaccines described herein as CIATEJ/OPKO/vaccines were generated in VERO cells and tested to generate anti-pandemic influenza AH1N1 The ability of antibodies. When an antigen produced by CIATEJ/OPKO in VERO cells is used as a vaccine in mice, it can induce antibody production. The titer of the antibody induced by the CIATEJ/OPKO antigen is greater than that of the commercially available vaccine. 159890.doc -69· 201249996 Figure 5 shows the total anti-influenza p/AHINl antibody detected by ELISA. The CIATEJ/OPKO/vaccine line is produced in VERO cells, TF = transfer factor. Immunization was performed in 7 to 8 week old B ALB/c female mice. The immune system is administered sublingually or intramuscularly. Figure 6A shows an anti-influenza p/AHINl IgM antibody detected in serum by ELISA. The CIATEJ/OPKO/vaccine line is produced in VERO cells, TF = transfer factor. Immunization was performed in 7-8 week old BALB/c female mice. The immune system is administered intramuscularly. When the transfer factor was added to the vaccine CIATEJ/OPKO, it increased the IgG antibody content by a factor of 5, and during the first week of immunization, the transfer factor increased the IgM antibody titer by a factor of three in the same manner. Figure 6B shows an anti-influenza p/AHIN1 IgG antibody detected in serum by ELISA. The CIATEJ/ΟΡΚΟ/vaccine line is produced in VERO cells, TF = transfer factor. Immunization was performed in 7-8 week old BALB/c female mice. The immune system is administered intramuscularly. Intramuscular administration of the vaccine CIATEJ/OPKO showed more induced IgGu antibodies than sublingual administration. Figure 7A shows an IgG2A antibody against pandemic influenza p/AHIN1 detected in serum by ELISA. The CIATEJ/OPKO/vaccine line is produced in VERO cells, TF = transfer factor. Immunization was performed in 7-8 week old BALB/C female mice. The immune system is administered sublingually or intramuscularly. Figure 7B shows the IgG2A dose dependency of anti-influenza p/AHINl detected in serum by ELISA. The CIATEJ/OPKO/vaccine line was produced in VERO cells, TF = transfer factor. Immunization was performed in 7-8 week old BALB/C female mice. The immune system is administered sublingually. Intramuscular administration of the vaccine CIATEJ/OPKO showed that the amount of IgA antibody induced was comparable to that of 159890.doc -70· 201249996 sublingual administration. Figure 8A shows an anti-influenza p/AHIN1 IgA antibody detected in the respiratory mucosa by ELISA. The CIATEJ/OPKO/vaccine line is produced in VERO cells, TF = transfer factor. Immunization was performed in 7-8 week old BALB/c female mice. The immune system is administered sublingually or intramuscularly (on day 21 after immunization). Figure 8B shows the IgA dose dependency of anti-influenza p/AHINl detected in the respiratory mucosa by ELISA. The CIATEJ/OPKO/vaccine line was produced in VERO cells, TF = transfer factor. Immunization was performed in 7-8 week old BALB/c female mice. The immune system is administered sublingually or intramuscularly (on day 21 after immunization). references

Magna Pure LC Total Nucleic Acid Isolation Kit. December 2008 edition, Kit for isolation of total viral nucleic acids from mammalian serum, plasma and whole blood, catalog number 03 038 505 001 °

Superscript III Platinum One Step Quantitative RT-PCR System. Catalog No. 1 1738-088.

World Health Organization. CDC protocol of Real time for Influenza A (H1N1). April 29, 2009 曰. CENAVECE-InDRE. Lineamientos para la red Nacional de laboratorios de salud piiblica e institutos nacionales de salud para vigilancia de influenza en Mexico 2009 o

Ammerman N., Beier M. and Azad F. 2008 Growth and

Maintenance of VERO Cell Lines. Curr Protoc Microbiol., 159890.doc -71- 201249996 Page 1-10.

Erlich Η., Mtiller Μ·, Helen Μ·, Tambyah P., Joukhadar Μ., Montomoli Μ., Fisher D., Berezuk D., Fritsch M., Low-Baselli A., Vartian N., Bobrovsky R., Borislava G., Pavlova, Pollabauer E., Kistner O. and Barrett N. 2008 A Clinical Trial of a Whole-Virus H5N1 Vaccine Derived from Cell Culture for the Baxter H5N1 Pandemic Influenza Vaccine Clinical Study Team. N. Engl. J. Med 358 ; 24., p. 2573-2584

Instituto de Medicina Tropical "Pedro Kouri".2003 · Manual de procedimientos para el diagnostico de laboratorio de la infecciones respiratyorias agudas de etiologia viral. Organizacion panamericana de la salud. Organizacion mundial de la salud o

Kistner O., Howard K., Spruth M., Wodal W., Brtihl P., Gerencer M., Crowe B., Savidis H., Livey I., Reiter M., Mayerhofer I., Tauer C., Grillberger L , Mundt G., Falkner F., and P. Noel. 2007 Cell culture (Vero) derived whole virus (H5N1) vaccine based on wild-type virus strain induces cross-protective immune responses. Baxter AG, Biomedical Research Center, Uferstrasse, Austria. Vaccine. 2007 10 ; 25(32). Pages 6028-6036.

Reina J. ' Fernandez V. ' Blanco I. y Munar M. 1997. Comparison of Madin-Darby Canine Kidney Cells (MDCK) 159890.doc • 71· 201249996 with a Green Monkey Continuous Cell Line (Vero) and Human Lung Embryonated Cells (MRC-5) in the Isolation of Influenza A Virus from Nasopharyngeal Aspirates by Shell Vial Culture. American Society for Microbiology Vol. 35, No. 7, pp. 1900-1901

Ryan John. Ph. D. Introduction to animal cell culture. Corning Incorporated Life Sciences 900 Chelmsford St. Lowell, Bulletin Technical. MA 01851, pp. 1-24.

Ryan John. Ph. D. Subculturing Monolayer cell cultures Protocol Corning Incorporated Life Sciences 900 Chelmsford St· Lowell, MA 01851, pp. 1-5. Sheets R. 2000 History and Characterization of the VERO Cell Line. Vaccines and Related Biological Products Advisory Committee Meeting to be held o All publications and patent applications cited in this specification are hereby incorporated by reference herein Publications or patent applications are expressly and individually indicated to be incorporated by reference in their entirety. Although the present invention has been described in detail by way of illustration and example of the embodiments of the invention, the invention may be It does not depart from the spirit or scope of the appended claims. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows an example of quantifying viral load by real-time PCR. Figure 2 shows the sample configuration of the AH1N1 real-time PCR assay. 159890.doc •73· 201249996 Figures 3A and 3B show a graphical view of the Light Cycler 2.0 Instant Temperature Cycler interface showing the fluorescence curve. Figure 3A) Clinical samples. Figure 3B) RNAseP (RP) assay Figure 4A shows the adaptation of the influenza virus HZFLUJAL in the VERO cell line. The virus replicates efficiently in VERO cells until 4 subcultures. Figure 4B shows the adaptation phase of the influenza virus HZFLUJAL in the MDCK cell line. After three subcultures, the influenza virus HZFLUJA was able to replicate efficiently in MDCK cells. Figure 5 shows the total anti-influenza p/AHINl antibody detected by ELISA. Figure 6A shows an anti-influenza p/AHINl IgM antibody detected in serum by ELISA. Figure 6B shows an anti-influenza p/AHINl IgG antibody detected in serum by ELISA.

I Figure 7A shows an anti-influenza p/AHINl IgG2A antibody detected in serum by ELISA. Figure 7B shows the IgG2A dose dependency of anti-influenza p/AHINl detected in serum by ELISA. Figure 8A shows an anti-influenza p/AHINl IgA antibody detected in the respiratory mucosa by ELISA. Figure 8B shows the IgA dose dependency of anti-influenza p/AHINl detected in the respiratory mucosa by ELISA. 159890.doc • 74- 201249996 Sequence Listing &lt;110&gt;&lt;120&gt; Novel Vaccine against Pandemic Influenza Virus A/H1N1

&lt;130&gt; P-75578-PC &lt;140&gt; 100140015 &lt;141&gt; 201M1-02 &lt;150&gt; 61/409,303 &lt;151&gt; 2010-11-02 &lt;160&gt; 52 &lt;170&gt; Patentln version 3.5

&Lt; 210 &gt; 1 &lt; 211 &gt; 2293 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 1 tatggagaga ataaaagaac tgagagatct aatgtcgcag tcccgcactc gcgagatact 60 cactaagacc actgtggacc atatggccat aatcaaaaag tacacatcag gaaggcaaga 120 gaagaacccc gcactcagaa tgaagtggat gatggcaatg agatacccaa ttacagcaga 180 caagagaata atggacatga ttccagagag gaatgaacaa ggacaaaccc tctggagcaa 240 aacaaacgat gctggatcag accgagtgat ggtatcacct ctggccgtaa catggtggaa 300 taggaatggc ccaacaacaa gtacagttca ttaccctaag gtatataaaa cttatttcga 360 aaaggtcgaa aggttgaaac atggtacctt cggccctgtc cacttcagaa atcaagttaa 420 aataaggagg agagttgata caaaccctgg ccatgcagat ctcagtgcca aggaggcaca 480 ggatgtgatt atggaagttg ttttcccaaa tgaagtgggg gcaagaatac tgacatcaga 540 gtcacagctg gcaataacaa aagagaagaa agaagagctc caggattgta aaattgctcc 600 Cttgatggtg gcgtacatgc tagaaagaga attggtccgt aaaacaaggt ttctcccagt 660 agccggcgga acaggcagtg tttatattga agtgttgcac ttaacccaag ggacgtgctg 720 ggagcagatg tacactccag gaggagaagt gagaaatgat gatgttgacc aaag tttgat 780 tatcgctgct agaaacatag taagaagagc agcagtgtca gcagacccat tagcatctct 840 cttggaaatg tgccacagca cacagattgg aggagtaagg atggtggaca tccttagaca 900 gaatccaact gaggaacaag ccgtagacat atgcaaggca gcaatagggt tgaggattag 960 ctcatctttc agttttggtg ggttcacttt caaaaggaca agcggatcat cagtcaagaa 1020 agaagaagaa gtgctaacgg gcaacctcca aacactgaaa ataagagtac atgaagggta 1080 tgaagaattc acaatggttg ggagaagagc aacagctatt ctcagaaagg caaccaggag 1140 attgatccag ttgatagtaa gcgggagaga cgagcagtca attgctgagg caataattgt 1200 ggccatggta ttctcacaag aggattgcat gatcaaggea gttaggggcg atctgaactt 1260 tgtcaatagg gcaaaccagc gactgaaccc catgcaccaa ctcttgaggc atttccaaaa 1320 agatgcaaaa gtgcttttcc agaactgggg aattgaatcc atcgacaatg tgatgggaat 1380 gatcggaata ctgcccgaca tgaccccaag cacggagatg tcgctgagag ggataagagt 1440 cagcaaaatg ggagtagatg aatactccag cacggagaga gtggtagtga gtattgaccg 1500 atttttaagg gttagagatc aaaga ^ ggaa cgtactattg.tctcccgaag aagtcagtga 1560 159890- sequence Listing .doc 201249996 aacgcaagga actgagaagt tgacaataac ttatt cgtca tcaatgatgt gggagatcaa 1620 tggccctgag tcagtgctag tcaacactta tcaatggata atcaggaact gggaaattgt 1680 gaaaattcaa tggtcacaag atcccacaat gttatacaac aaaatggaat ttgaaccatt 1740 tcagtctctt gtccctaa ^ g caaccagaag ccggtacagt ggattcgtaa ggacactgtt 1800 ccagcaaatg cgggatgtgc ttgggacatt tgacactgtc caaataataa aacttctccc 1860 ctttgctgct gctccaccag aacagagtag gatgcaattt tcctcattga ctgtgaatgt 1920 gagaggatca gggttgagga tactggtaag aggcaattct ccagtattca attacaacaa 1980 ggcaaccaaa cgacttacag ttcttggaaa ggatgcaggt gcattgactg aagatccaga 2040 tgaaggcaca tctggggtgg agtctgctgt cctgagagga tttctcattt tgggcaaaga 2100 agacaagaga tatggcccag cattaagcat caatgaactg agcaatcttg caaaaggaga 2160 gaaagctaat gtgctaattg ggcaagggga cgtagtgttg gtaatgaaac gaaaacggga 2220 ctctagcata cttactgaca gccagacagc gaccaaaaga attcggatgg ccatcaatta 2280 gtgtcgaatt gtt 2293

&Lt; 210 &gt; 2 &lt; 211 &gt; 2303 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 2 ttgaatggat gtcaatccga ctctactttt cctaaaaatt ccagcgcaaa atgccataag 60 caccacattc ccttatactg gagatcctcc atacagccat ggaacaggaa caggatacac 120 catggacaca gtaaacagaa cacaccaata ctcagaaaag ggaaagtgga cgacaaacac 180 agagactggt gcaccccagc tcaacccgat tgatggacca ctacctgagg ataatgaacc 240 aagtgggtat gcacaaacag actgtgttct agaggctatg gctttccttg aagaatccca 300 cccaggaata tttgagaatt catgccttga aacaatggaa gttgttcaac aaacaagggt 360 agataaacta actcaaggtc gccagactta tgattggaca ttaaacagaa atcaaccggc 420 agcaactgca ttggccaaca ccatagaagt ctttagatcg aatggcctaa cagctaatga 480 gtcaggaagg ctaatagatt tcttaaagga tgtaatggaa tcaatgaaca aagaggaaat 540 agagataaca acccactttc aaagaaaaag gagagtaaga gacaacatga ccaagaagat 600 Ggtcacgcaa agaacaatag ggaagaaaaa acaaagactg aataagagag gctatctaat 660 aagagcactg acattaaata cgatgaccaa agatgcagag agaggcaagt taaaaagaag 720 ggctatcgca acacctggga tgcagattag aggtttcgta tactttgttg aaac tttagc 780 taggagcatt tgcgaaaagc ttgaacagtc tgggctccca gtagggggca atgaaaagaa 840 ggccaaactg gcaaatgttg tgagaaagat gatgactaat cagagatttc 900 tttcacaatc actggggaca acactaagtg gaatgaaaat caaaatcctc gaatgttcct 960 ggcgatgatt acatatatca ccagaaatca acccgagtgg ttcagaaaca tcctgagcat 1020 ggcacccata atgttctcaa acaaaatggc aagactaggg aaagggtaca tgttcgagag 1080 taaaagaatg aagattcgaa cacaaatacc agcagaaatg ctagcaagca ttgacctgaa 1140 gtacttcaat gaatcaacaa agaagaaaat tgagaaaata aggcctcttc taatagatgg tcacaagaca 1200 cacagcatca ctgagtcctg ggatgatgat gggcatgttc aacatgctaa gtacggtctt 1260 -2- 159890 · sequence Listing .doc 201249996 gggagtctcg atactgaatc ttggacaaaa gaaatacacc aagacaatat actggtggga 1320 tgggctccaa tcatccgacg attttgctct catagtgaat gcaccaaacc atgagggaat 1380 acaagcagga gtggacagat tctacaggac ctgcaagtta gtgggaatca acatgagcaa 1440 aaagaagtcc tatataaata agacagggac atttgaattc acaagctttt tttatcgcta 1500 tggatttgtg gctaatttta gcatggagct acccagcttt Ggagtgtctg gagtaaatga 1560 atcagctgac atga£tattg gagtaacagt gataaagaac aacatgataa acaatgacct 1620 tggacctgca acggcccaga tggctcttca attgttcatc aaagactaca gatacacata 1680 taggtgccat aggggagaca cacaaattca gacgagaaga tcatttgagt taaagaagct 1740 gtgggatcaa acccaatcaa aggtagggct attagtatca gatggaggac caaacttata 1800 caatatacgg aatcttcaca ttcctgaagt ctgcttaaaa tgggagctaa tggatgatga 1860 ttatcgggga agactttgta atcccctgaa tccctttgtc agtcataaag agattgattc 1920 tgtaaacaat gctgtggtaa tgccagccca tggtccagcc aaaagcatgg aatatgatgc 1980 cgttgcaact acacattcct ggattcccaa gaggaatcgt tctattctca acacaagcca 2040 aaggggaatt cttgaggatg aacagatgta ccagaagtgc tgcaatctat tcgagaaatt 2100 tttccctagc agttcatata ggagaccggt tggaatttct agcatggtgg aggccatggt 2160 gtctagggcc cggattgatg ccagggtcga cttcgagtct ggacggatca agaaagaaga 2220 gttctctgag atcatgaaga tctgttccac cattgaagaa ctcagacggc aaaaataatg 2280 aatttaactt gtccttcatg aaa 2303

&Lt; 210 &gt; 3 &lt; 211 &gt; 2172 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 3 tccaaaatgg aagactttgt gcgacaatgc ttcaatccaa tgatcgtcga gcttgcggaa 60 aaggcaatga aagaatatgg ggaagatccg aaaatcgaaa ctaacaagtt tgctgcaata 120 tgcacacatt tggaagtttg tttcatgtat tcggatttcc atttcatcga cgaacggggt 180 gaatcaataa ttgtagaatc tggtgacccg aatgcactat tgaagcaccg atttgagata 240 attgaaggaa gagaccgaat catggcctgg acagtggtga acagtatatg taacacaaca 300 ggggtagaga agcctaaatt tcttcctgat ttgtatgatt acaaagagaa ccggttcatt 360 gaaattggag taacacggag ggaagtccac atatattacc tagagaaagc caacaaaata 420 aaatctgaga agacacacat tcacatcttt tcattcactg gagaggagat ggccaccaaa 480 gcggactaca cccttgac £ a agagagcagg gcaagaatca aaactaggct tttcactata 540 agacaagaaa tggccagtag gagtctatgg gattcctttc gtcagtccga Aagaggcgaa 600 gagacaattg aagaaaaatt tgagattaca ggaactatgc gcaagcttgc cgaccaaagt 660 ctcccaccga acttctccag ccttgaaaac tttagagcct atgtagatgg attcgagccg 720 aacggctgca ttgagggcaa gctttcccaa atgtcaaaag aagtgaacgc caa aattgaa 780 ccattcttga ggacgacacc acgccccctc agattgcctg atgggcctct ttgccatcag 840 cggtcaaagt tcctgctgat ggatgctctg aaattaagta ttgaagaccc gagtcacgag 900 SSgS ^ gggaa taccactata tgatgcaatc aaatgcatga agacattctt tggctggaaa 960 159890 · Sequence Listing .doc 201249996 gagcctaaca tagtcaaacc acatgagaaa ggcataaatc ccaattacct catggcttgg 1020 aagcaggtgc tagcagagct acaggacatt gaaaatgaag agaagatccc aaggacaaag 1080 aacatgaaga gaacaagcca attgaagtgg gcactcggtg aaaatatggc accagaaaaa 1140 gtagactttg atgactgcaa agatgttgga gaccttaaac agtatgacag tgatgagcca 1200 gagcccagat ctctagcaag ctgggtccaa aatgaattca ataaggcatg tgaattgact 1260 gattcaagct ggatagaact tgatgaaata ggagaagatg ttgccccgat tgaacatatc 1320 gcaagcatga ggaggaacta ttttacagca gaagtgtccc actgcagggc tactgaatac 1380 ataatgaagg gagtgtacat aaatacggcc ttgctcaatg catcctgtgc agccat £ gat 1440 gactttcagc tgatcccaat gataagcaaa tgtaggacca aagaaggaag acggaaaaca 1500 aacctgtatg ggttcattat aaaaggaagg Tctcatttga gaaatgatac tgatgtggtg 1560 aactttgtaa gtatggagtt ctcactcact ga cccgagac tggagccaca caaatgggaa 1620 aaatactgtg ttcttgaaat aggagacat £ ctcttgagga ctgcgatagg ccaagtgtcg 1680 aggcccatgt tcctatatgt gagaaccaat ggaacctcca agatcaagat gaaatggggc 1740 atggaactga ggcgctgcct tcttcagtct cttcagcaga ttgagagcat gattgaggcc 1800 gagtcttctg tcaaagagaa agacatgacc aaggaattct ttgaaaacaa atcggaaaca 1860 tggccaatcg gagagtcacc caggggagtg gaggaaggct ctattgggaa agtgtgcagg 1920 accttactgg caaaatctgt attcaacagt ctatatgcgt ctccacaact tgaggggttt 1980 tcggctgaat cgagaaaatt gcttctcatt gttcaggcac ttagggacaa cctggaacct 2040 ggaaccttcg atcttggggg gctatatgaa gcaatcgagg agtgcctgat taatgatccc 2100 tgggttttgc ttaatgcatc ttggttcaac tccttcctca cacatgcact gaagtagttg 2160 tggcaatgct ac 2172 &lt; 210 &gt; 4 &lt; 211 &gt; 1751 &lt; 212 &gt; IM &lt; 213 &gt; H1N1 porcine epidemic sexy f virus &lt; 400 &gt; 4 agcaaaagca ggggaaaaca aaagcaacaa aaatgaaggc aatactagta gttctgctat 60 acacatttgc aaccgcaaat gcagacacat tatgtatagg ttatcatgcg aacaattcaa 120 cagacactgt agacacagta ctagaaaaga atgtaacagt aacacactct gttaaccttc 1 80 tagaagacaa gcataacggg aaactatgca aactaagagg ggtagcccca ttgcatttgg 240 gtaaatgtaa cattgctggc tggatcctgg gaaatccaga gtgtgaatca ctctccacag 300 caagctcatg gtcctacatt gtggaaacat ctagttcaga caatggaacg tgttacccag 360 gagatttcat cgattatgag gagctaagag agcaattgag-ctcagtgtca tcatttgaaa 420 ggtttgagat attccccaag acaagttcat ggcccgatca tgactcgaac aaaggtgtaa 480 cggcagcatg tcctcatgct ggagcaaaaa gcttctacaa aaatttaata tggctagtta 540 aaaaaggaaa ttcataccca aagctcagca aatcctacat taatgataaa gggaaagaag 600 tcctcgtgct atggggcatt caccatccat ctactagtgc tgaccaacaa agtctctatc 660 agaatgcaga tgcatatgtt catcaagata cagcaagaag ttcaagccgg 720 aaatagcaat aagacccaaa gtgagggaac gagaagggag aatgaactat tactggacac 780 • 4- 159890- sequence Listing doc acgattgcaa tacaacttgt cagacaccca tttgtgggga 201249996 tagtagagcc gggagacaaa ataacattcg aagcaactgg aaatctagtg gtaccgagat 840 atgcattcgc aatggaaaga aatgctggat ctggtattat catttcagat acaccagtcc 900 aaggtgctat aaacaccagc Ctcccatttc 960 agaatataca tccgatcaca attggaaaat gtccaaaata tgtaa aaagc acaaaattga 1020 gactggccac aggattgagg aatgtcccgt ctattcaatc tagaggccta tttggggcca 1080 ttgccggttt cattgaaggg gggtggacag ggatggtaga tggatggtac ggttatcacc 1140 atcaaaatga gcaggggtca ggatatgcag ccgacctgaa gagcacacag aatgccattg 1200 acgagattac taacaaagta aattctgtta ttgaaaagat gaatacacag ttcacagcag 1260 taggtaaaga gttcaaccac ctggaaaaaa gaatagagaa tttaaataaa aaagttgatg 1320 atggtttcct ggacatttgg acttacaatg ccgaactgtt ggttctattg gaaaatgaaa 1380 gaactttgga ctaccacgat tcaaatgtga agaacttata tgaaaaggta agaagccagt 1440 taaaaaacaa tgccaaggaa attggaaacg gctgctttga attttaccac aaatgcgata 1500 acacgtgcat ggaaagtgtc aaaaatggga cttatgacta cccaaaatac tcagaggaag 1560 caaaattaaa cagagaagaa atagatgggg taaagctgga atcaacaagg atttaccaga 1620 ttttggcgat ctattcaact gtcgccagtt cattggtact ggtagtctcc ctgggggcaa 1680 tcagtttctg gatgtgctct aatgggtctc tacagtgtag aatatgtatt taacattagg 1740 atttcagaag c 1751

&Lt; 210 &gt; 5 &lt; 211 &gt; 1546 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 5 atcaatcact gagtgacatc gaagccatgg cgtctcaagg caccaaacga tcatatgaac 60 aaatggagac tggtggggag cgccaggatg ccacagaaat cagagcatct gtcggaagaa 120 tgattggtgg aatcgegaga ttctacatcc aaatgtgcac tgaactcaaa ctcagtgatt 180 atgatggacg actaatccag aatagcataa caatagagag gatggtgctt tctgcttttg 240 atgagagaag aaataaatac ctagaagagc atcccagtgc tgggaaggac cctaagaaaa 300 caggaggacc catatataga agaatagacg gaaagtggat gagagaactc atcctttatg 360 acaaagaaga aataaggaga gtttggcgcc aagcaaacaa tggcgaagat gcaacagcag 420 gtcttactca tatcatgatt tggcattcca acctgaatga tgccacatat cagagaacaa 480 gagcgcttgt tcgcaccgga atggatccca gaatgtgctc tctaatgcaa ggttcaacac 540 ttcccagaag gtctggtgcc gcaggtgctg cggtgaaagg agttggaaca atagcaatgg 600 Agttaatcag aatgatcaaa cgtggaatca atgaccgaaa tttctggagg ggtgaaaatg 660 gacgaaggac aagggttgct tatgaaagaa tgtgcaatat cctcaaagga aaatttcaaa 720 cagctgccca gagggcaatg atggatcaag taagagaaag tcgaaaccca ggaa acgctg 780 agattgaaga cctcattttc ctggcacggt cagcactcat tctgagggga tcagttgcac 840 ataaatcctg cctgcctgct tgtgtgtatg ggcttgcagt agcaagtggg catgactttg 900 aaagggaagg gtactcactg gtcgggatag acccattcaa attactccaa aacagccaag 960 tggtcagcct gatgagacca aatgaaaacc cagctcacaa gagtcaattg gtgtggatgg 1020 -5- 159890- Sequence Listing .doc 201249996 catgccactc tgctgcattt gaagatttaa gagtatcaag tttcataaga ggaaagaaag 1080 tgattccaag aggaaagctt tccacaagag gggtccagat tgcttcaaat gagaatgtgg 1140 aaaccatgga ctccaatacc ctggaactaa gaagcagata ctgggccata aggaccagga 1200 gtggaggaaa taccaatcaa caaaaggcat ccgcaggcca gatcagtgtg cagcctacat 1260 tctcagtgca gcgaaatctc ccttttgaaa gagcaaccgt tatggcagca ttcagcggga 1320 acaatgaagg acggacatcc gacatgcgaa cagaagttat aagaatgatg gaaagtgcaa 1380 agccagaaga tttgtccttc caggggcggg gagtcttcga gctctcggac gaaaaggcaa 1440 cgaacccgat cgtgccttcc tttgacatga gtaatgaagg gtcttatttc ttcggagaca 1500 atgcagagga gtatgacagt tgaggaaaaa tacccttgtt Tctact 1546 &lt;210&gt; 6 &lt;211&gt; 1410 &lt;212&gt; Leg & l t; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 6 atgaatccaa accaaaagat aataaccatt ggttcggtct gtatgacgat tggaatggct 60 aacttaatat tacaaattgg aaacataatc tcaatatgga ttagccactc aattcaactt 120 gggaatcaaa atcagattga aacatgcaat caaagcgtca ttacttatga aaacaacact 180 tgggtaaatc agacatatgt taacatcagc aacaccaact ttgctgctgg acagtcagtg 240 gtttccgtga aattagcggg caattcctct ctctgccctg ttagtggatg ggctatatac 300 agtaaagaca acagtataag aatcggttcc aagggggatg tgtttgtcat aagggaacca 360 ttcatatcat gctccccctt ggaatgcaga accttcttct tgactcaagg ggccttgcta 420 aatgacaaac attccaatgg aaccattaaa gacaggagcc catatcgaac cctaatgagc 480 tgtcctattg gtgaagttcc ctctccatac aactcaagat ttgagtcagt cccagacaat 600 ggggcagtgg ctgtgttaaa gtacaacggc ataataacag acactatcaa gagttggaga 660 aacaatatat tgagaacaca agagtctgaa tgtgcatgtg taaatggttc ttgctttact cgcttggtca 540 gcaagtgctt gtcatgatgg catcaattgg ctaacaattg gaatttctgg 720 gtaatgaccg Atggaccaag tgatggacag gcctcataca agatcttcag aatagaaaa £ 780 ggaaagatag tcaaatcagt cgaaatgaat gccc ctaatt attactatga ggaatgctcc 840 tgttatcctg attctagtga aatcacatgt gtgtgcaggg ataactggca tggctcgaat 900 cgaccgtggg tgtctttcaa ccagaatctg gaatatcaga taggatacat atgcagtggg 960 attttcggag acaatccacg ccctaatgat aagacaggca gttgtggtcc agtatcgtct 1020 aatggagcaa atggagtaaa aggattttca ttcaaatacg gcaatggtgt ttggataggg 1080 agaactaaaa gcattagttc aagaaacggt tttgagatga tttgggatcc gaacggatgg 1140 actgggacag acaataactt ctcaataaag caagatatcg taggaataaa tgagtggtca 1200 ggatatagcg £ gagttttgt tcagcatcca Gaactaacag ggctggattg tataagacct 1260 tgcttctggg ttgaactaat ca£a£ggcga cccaaagaga acacaatctg gactagcggg 1320 agcagcatat ccttttgtgg tgtaaacagt gacactgtgg gttggtcttg £ccagacg£t 1380 gctgagttgc catttaccat tgacaagtaa 1410 159890·sequence table.doc 201249996

&Lt; 210 &gt; 7 &lt; 211 &gt; 1368 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 7 atgagtcttc taaccgaggt cgaaacgtac gttctttcta tcatcccgtc aggccccctc 60 aaagccgaga tcgcgcagag actggaaagt gtctttgcag gaaagaacac agatcttgag 120 gctctcatgg aatggctaaa gacaagacca atcttgtcac ctctgactaa gggaatttta 180 ggatttgtgt tcacgctcac cgtgcccagt gagcgaggac tgcagcgtag acgctttgtc 240 caaaatgccc taaatgggaa tggggacccg aacaacatgg atagagcagt taaactatac 300 aagaagctca aaagagaaat aacgttccat ggggccaagg aggtgtcact aagctattca 360 actggtgcac ttgccagttg catgggcctc atatacaaca ggatgggaac agtgaccaca 420 gaagctgctt ttggtctagt gtgtgccact tgtgaacaga ttgctgattc acagcatcgg 480 tctcacagac agatggctac taccaccaat ccactaatca ggcatgaaaa cagaatggtg 540 ctggctagca ctacggcaaa ggctatggaa cagatggctg gatcgagtga acagaaagga 600 Gaagacgtcc aaaagctggc agaagagctg caaagcaaca ttggagtatt gagatctctt 660 ggagcaagtc aaaagagtgg ggaaggaatt gcaaaggatg taatggaagt gctaaagcag 720 agctctatgg gaaattcagc tcttgtgaag aaatatctat gctcgaacca tttc agattc 780 tttcaatttg ttcttttatc ttatcagctc tccatttcat ggcttggaca aggcattatc 840 aaaaaagagg agagaaaaga aaaggtccaa acaaagagac aaacagagag gtatcaattt 900 gacacagtta ccaaaaagaa atccaggcca aagaaacaaa ggaagtactc tctgacaaca 960 agggtgacca caaggggctt tctgccggcg gaggccatgg aggttgctaa tcagactagg 1020 cagatggtac atgcaatgag aactattggg actcatccta gctccagtgc tggtctgaaa 1080 gatgaccttc ttgaaaattt gcaggcctac cagaagcgaa tgggagtgca gatgcagcga 1140 ttcaagtcct ctcgtcattg cagcaatatc attgggatct tgcacctgat attgtggatt 1200 actgatcgtc tttttttcaa atgtatttat cgtcgcttta aatacggttt gaaaagaggg 1260 ccttctacgg aaggagtgcc tgagtccatg agggaagaat atcaacagga acagcagagt 1320 gctgtggatg ttgacgatgg tcattttgtc aacatagagc tagagtaa 1368

&Lt; 210 &gt; 8 &lt; 211 &gt; 851 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 8 aacataatgg actccaacac catgtcaagc tttcaggtag actgtttcct ttggcatatc 60 cgcaagcgat ttgcagacaa tggattgggt gatgccccat tccttgatcg gctccgccga 120 gatcaaaagt ccttaaaagg aagaggcaac acccttggcc tcgatatcga aacagccact 180 cttgttggga aacaaatcgt ggaatggatc ttgaaagagg aatccagcga gacacttaga 240 atgacaattg catctgtacc tacttcgcgc tacctttctg acatgaccct cgaggaaatg 300 tcacgagact ggttcatgct catgcctagg caaaagataa taggccctct ttgcgtgcga 360 ttggaccagg cgatcatgga aaagaacata gtactgaaag cgaacttcag tgtaatcttt 420 aaccgattag agaccttgat actactaagg gctttcactg aggagggagc aatagttgga 480 159890- sequence Listing .d〇c 201249996 gaaatttcac cattaccttc tcttccagga catacttatg aggatgtcaa Aaatgcaatt 540 ggggtcctca tcggaggact tgaatggaat ggtaacacgg ttcgagtctc tgaaaatata 600 cagagattcg cttggagaaa ctgtgatgag aatgggagac cttcactacc tccagagcag 660 aaatgaaaag tggcgagagc aattgggaca gaaatttgag gaaataaggt ggttaattga 720 agaaatgcgg cacagattga aagcg Acaga gaatagtttc gaacaaataa catttatgca 780 agccttacaa ctactgcttg aagtagaaca agagataaga gctttctcgt ttcagcttat 840 ttaactgata a 851

&Lt; 210 &gt; 9 &lt; 211 &gt; 2293 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 9 tatggagaga ataaaagaac tgagagatct aatgtcgcag tcccgcactc gcgagatact 60 cactaagacc actgtggacc atatggccat aatcaaaaag tacacatcag gaasgcaaga 120 gaagaacccc gcactcagaa tgaagtggat gatggcaatg agatacccaa ttacagcaga 180 caagagaata atggacatga ttccagagag gaatgaacaa ggacaaaccc tctggagcaa 240 aacaaacgat gctggatcag accgagtgat ggtatcacct ctggccgtaa catggtggaa 300 taggaatggc ccaacaacaa gtacagttca ttaccctaag gtatataaaa cttatttcga 360 aaaggtcgaa aggttgaaac atggtacctt cggccctgtc cacttcagaa atcaagttaa 420 aataaggagg agagttgata caaaccctgg ccatgcagat ctcagtgcca aggaggcaca 480 ggatgtgatt atggaagttg ttttcccaaa tgaagtgggg gcaagaatac tgacatcaga 540 gtcacagctg gcaataacaa aagagaagaa agaagagctc caggattgta aaattgctcc 600 Cttgatggtg gcgtacatgc tagaaagaga attggtccgt aaaacaaggt ttctcccagt 660 agccggcgga acaggcagtg tttatattga agtgttgcac ttaacccaag ggacgtgctg 720 ggagcagatg tacactccag gaggagaagt gagaaatgat gatgttgacc aaag tttgat 780 tatcgctgct agaaacatag taagaagagc agcagtgtca gcagacccat tagcatctct 840 cttggaaatg tgccacagca cacagattgg aggagtaagg atggtggaca tccttagaca 900 gaatccaact gaggaacaag ccgtagacat atgcaaggca gcaatagggt tgaggattag 960 ctcatctttc agttttggtg ggttcacttt caaaaggaca agcggatcat cagtcaagaa 1020 agaagaagaa gtgctaacgg gcaacctcca aacactgaaa ataagagtac atgaagggta 1080 tgaagaattc acaatggttg ggagaagagc aacagctatt ctcagaaagg caaccaggag 1140 attgatccag ttgatagtaa gcgggagaga cgagcagtca attgctgagg caataattgt 1200 ggccatggta ttctcacaag aggattgcat gatcaaggca gttaggggcg atctgaactt 1260 tgtcaatagg gcaaaccagc gactgaaccc catgcaccaa ctcttgaggc atttccaaaa 1320 agatgcaaaa gtgcttttcc agaactgggg aattgaatcc atcgacaatg tgatgggaat 1380 gatcggaata ctgcccgaca tgaccccaag cacggagatg tcgctgagag ggataagagt 1440 cagcaaaatg ggagtagatg aatactccag cacggagaga gtggtagtga gtattgaccg 1500 atttttaagg gttagagatc aaagagggaa cgtactattg tctcccgaag aagtcagtga 1560 aacgcaagga actgagaagt tgacaataac ttattcgtca tcaatgatgt gggagatcaa 162 0159890 * Sequence Listing .doc 201249996 tggccctgag tcagtgctag tcaacactta tcaatggata atcaggaact gggaaattgt 1680 gaaaattcaa tggtcacaag atcccacaat gttatacaac aaaatggaat ttgaaccatt 1740 tcagtctctt gtccctaagg caaccagaag ccggtacagt ggattcgtaa ggacactgtt 1800 ccagcaaatg cgggatgtgc ttgggacatt tgacactgtc caaataataa aacttctccc 1860 ctttgctgct gctccaccag aacagagtag gatgcaattt tcctcattga ct £ tgaatgt 1920 gagaggatca gggttgagga tactggtaag aggcaattct cca £ tattca attacaacaa 1980 ggcaaccaaa cgacttacag ttcttggaaa ggatgcaggt gcattgactg aagatccaga 2040 tgaaggcaca tctggggtgg agtctgctgt cctgagagga tttctcattt tgggcaaaga 2100 agacaagaga tatggcccag cattaagcat caatgaactg agcaatcttg caaaaggaga 2160 gaaagctaat gtgctaattg £ gcaagggga cgtagtgttg gtaatgaaac gaaaacggga 2220 ctctagcata cttactgaca gccagacagc gaccaaaaga attcggatgg ccatcaatta 2280 gtgtcgaatt gtt 2293

&Lt; 210 &gt; 10 &lt; 211 &gt; 2303 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 10 ttgaatggat gtcaatccga ctctactttt cctaaaaatt ccagcgcaaa atgccataag 60 caccacattc ccttatactg gagatcctcc atacagccat ggaacaggaa caggatacac 120 catggacaca gtaaacagaa cacaccaata ctcagaaaag ggaaagtgga cgacaaacac 180 agagactggt gcaccccagc tcaacccgat tgatggacca ctacctgagg ataatgaacc 240 aagtgggtat gcacaaacag actgtgttct agaggctatg gctttccttg aagaatccca 300 cccaggaata tttgagaatt catgccttga aacaatggaa gttgttcaac aaacaagggt 360 agataaacta actcaaggtc gccagactta tgattggaca ttaaacagaa atcaaccggc 420 agcaactgca ttggccaaca ccatagaagt ctttagatcg aatggcctaa cagctaatga 480 gtcaggaagg ctaatagatt tcttaaagga tgtaatggaa tcaatgaaca aagaggaaat 540 agagataaca acccactttc aaagaaaaag gagagtaaga gacaacatga ccaagaagat 600 Ggtcacgcaa agaacaatag ggaagaaaaa acaaagactg aataagagag gctatctaat 660 aagagcactg acattaaata cgatgaccaa agatgcagag agaggcaagt taaaaagaag 720 ggctatcgca acacctggga tgcagattag aggtttcgta tactttgttg aa actttagc 780 taggagcatt tgcgaaaagc ttgaacagtc tgggctccca gtagggggca atgaaaagaa 840 ggccaaactg gcaaatgttg tgagaaagat gatgactaat cagagatttc 900 tttcacaatc actggggaca acactaagtg gaatgaaaat caaaatcctc gaatgttcct 960 ggcgatgatt acatatatca ccagaaatca acccgagtgg ttcagaaaca tcctgagcat 1020 ggcacccata atgttctcaa acaaaatggc aagactaggg aaagggtaca tgttcgagag 1080 taaaagaatg aagattcgaa cacaaatacc agcagaaatg ctagcaagca ttgacctgaa 1140 gtacttcaat gaatcaacaa agaagaaaat tgagaaaata aggcctcttc taatagatgg tcacaagaca 1200 cacagcatca ctgagtcctg ggatgatgat gggcatgttc aacatgctaa gtacggtctt 1260 gggagtctcg atactgaatc ttggacaaaa gaaatacacc aagacaatat actggtggga 1320 159890 · sequence Listing .doc 201249996 tgggctccaa tcatcc £ acg attttgctct catagtgaat gcaccaaacc atgagggaat 1380 acaagcagga gtggacagat tctacaggac ctgcaagtta gtgggaatca acatgagcaa 1440 aaagaagtcc tatataaata agacagggac atttgaattc acaagctttt tttatcgcta 1500 tggatttgtg gctaatttta gcatggagct acccagcttt ggagtgtctg Gagtaaatga 1560 atcagctgac atgagtattg gagtaacagt g ataaagaac aacatgataa acaatgacct 1620 tggacctgca acggcccaga tggctcttca attgttcatc aaagactaca gatacacata 1680 taggtgccat aggggagaca cacaaattca gacga £ aaga tcatttgagt taaagaagct 1740 gtgggatcaa acccaatcaa aggtagggct attagtatca gatggaggac caaacttata 1800 caatatacgg aatcttcaca ttcctgaagt ctgcttaaaa tgggagctaa tggatgatga 1860 ttatcgggga agactttgta atcccctgaa tccctttgtc agtcataaag agattgattc 1920 tgtaaacaat gctgtggtaa tgccagccca tggtccagcc aaaagcatgg aatatgatgc 1980 cgttgcaact acacattcct ggattcccaa gaggaatcgt tctattctca acacaagcca 2040 aaggggaatt cttgaggatg aacagatgta ccagaagtgc tgcaatctat tcgagaaatt 2100 tttccctagc agttcatata ggagaccggt tggaatttct agcatggtgg aggccatggt 2160 gtctagggcc cggattgatg ccagggtcga cttcgagtct ggacggatca agaaagaaga 2220 gttctctgag atcatgaaga tctgttccac cattgaagaa ctcagacggc aaaaataatg 2280 aatttaactt gtccttcatg aaa 2303

&Lt; 210 &gt; 11 &lt; 211 &gt; 2172 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 11 tccaaaatgg aagactttgt gcgacaatgc ttcaatccaa tgatcgtcga gcttgcggaa 60 aaggcaatga aagaatatgg ggaagatccg aaaatcgaaa ctaacaagtt tgctgcaata 120 tgcacacatt tggaagtttg tttcatgtat tcggatttcc atttcatcga cgaacggggt 180 gaatcaataa ttgtagaatc tggtgacccg aatgcactat tgaagcaccg atttgagata 240 attgaaggaa gagaccgaat catggcctgg acagtggtga acagtatatg taacacaaca 300 ggggtagaga agcctaaatt tcttcctgat ttgtatgatt acaaagagaa ccggttcatt 360 gaaattggag taacacggag ggaagtccac atatattacc tagagaaagc caacaaaata 420 aaatctgaga agacacacat tcacatcttt tcattcactg gagaggagat ggccaccaaa 480 gcggactaca cccttgacga agagagcagg gcaa £ aatca aaactaggct tttcactata 540 agacaagaaa tggccagtag gagtctatgg gattcctttc gtcagtccga Aagaggcgaa 600 gagacaattg aagaaaaatt tgagattaca ggaactatgc gcaagcttgc cgaccaaagt 660 ctcccaccga acttctccag ccttgaaaac tttagagcct atgtagatgg attcgagccg 720 aacggctgca ttgagggcaa gctttcccaa atgtcaaaag aagtgaacgc c aaaattgaa 780 ccattcttga ggacgacacc acgccccctc agattgcctg atgggcctct ttgccatcag 840 cggtcaaagt tcctgctgat ggatgctctg aaattaagta ttgaagaccc gagtcacgag 900 egS £ agggaa taccactata tgatgcaatc aaatgcatga agacattctt tggctggaaa 960 gagcctaaca tagtcaaacc acatgagaaa ggcataaatc ccaattacct catggcttgg 1020 • 10 · 159890 · Sequence Listing .doc 201249996 aagcaggtgc tagcagagct acaggacatt gaaaatgaag agaagatccc aaggacaaag 1080 aacatgaaga gaacaagcca attgaagtgg gcactcggtg aaaatatggc accagaaaaa 1140 gtagactttg atgactgcaa agatgttgga gaccttaaac agtatgacag tgatgagcca 1200 gagcccagat ctctagcaag ctgggtccaa aatgaattca ataaggcatg tgaattgact 1260 gattcaagct ggatagaact tgatgaaata ggagaagatg ttgccccgat tgaacatatc 1320 gcaagcatga ggaggaacta ttttacagca gaagtgtccc actgcagggc tactgaatac 1380 ataatgaagg gagtgtacat aaatacggcc ttgctcaatg catcctgtgc agccatggat 1440 gactttcagc tgatcccaat gataagcaaa tgtaggacca aagaaggaag acggaaaaca 1500 aacctgtatg ggttcattat Aaaaggaagg tctcatttga gaaatgatac tgatgtggtg 1560 aactttgtaa gtatggagtt ct cactcact gacccgagac tggagccaca caaatgggaa 1620 aaatactgtg ttcttgaaat aggagacatg ctcttgagga ctgcgatagg ccaagtgtcg 1680 aggcccatgt tcctatatgt gagaaccaat ggaacctcca agatcaagat gaaatggggc 1740 atggaactga ggcgctgcct tcttcagtct cttcagcaga ttgagagcat gattgaggcc 1800 gagtcttctg tcaaagagaa agacatgacc aaggaattct ttgaaaacaa atcggaaaca 1860 tggccaatcg gagagtcacc caggggagtg gaggaaggct ctattgggaa agtgtgcagg 1920 accttactgg caaaatctgt attcaacagt ctatatgcgt ctccacaact tgaggggttt 1980 tcggctgaat cgagaaaatt gcttctcatt gttcaggcac ttagggacaa cctggaacct 2040 ggaaccttcg atcttggggg gctatatgaa gcaatcgagg agtgcctgat taatgatccc 2100 tgggttttgc ttaatgcatc ttggttcaac tccttcctca cacatgcact gaagtagttg 2160 tggcaatgct ac 2172 &lt; 210 &gt; 12 &lt; 211 &gt; 1735 &lt; 212 &gt; mk &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 12 aaaagcaaca aaagatgaag gcaatactag tagttctgct atatacattt gcaaccgcaa 60 atgcagacac attatgtata ggttatcatg cgaacaattc aacagacact gtagacacag 120 tactagaaaa gaatgtaaca gtaacacact ctgttaacct tctagaaga c aagcataacg 180 ggaaactatg caaactaaga ggggtagccc cattgcattt gggtaaatgt aacattgctg 240 gctggatcct gggaaatcca gagtgtgaat cactctccac agcaagctca tggtcctaca 300 ttgtggaaac atctagttca gacaatggaa cgtgttaccc aggagatttc atcgattatg 360 aggagctaag agagcaattg agctcagtgt catcatttga aaggtttgag atattcccca 420 agacaagttc atggcccaat catgactcga acaaaggtgt aacggcagca tgtcctcatg 480 ctggagcaaa aagcttctac aaaaatttaa tatggctagt taaaaaagga aattcatacc 540 caaagctcag caaatcctac attaatgata aagggaaaga agtcctcgtg ctatggggca 600 ttcaccatcc atctactagt gctgaccaac aaagtctcta tcagaatgca gatgcatatg 660 tttttgtggg gtcatcaaga tacagcaaga agttcaagcc ggaaatagca ataagaccca 720 aagtgaggga tcaagaaggg agaatgaact attactggac actagtagag ccgggagaca 780 aaataacatt cgaagcaact ggaaatctag tggtaccgag atatgcattc gcaatggaaa 840 • 11 · 159890- sequence table 201249996 gaaatgctgg atctggtatt atcatttcag atacaccagt ccacgattgc aatacaactt 900 gtcagacacc caagggtgct ataaacacca gcctcccatt tcagaatata Catccgatca 960 caattggaaa atgtccaaaa tatgtaaaaa gcaca Aaatt gagactggcc acaggattga 1020 ggaatgtccc gtctattcaa tctagaggcc tatttggggc cattgccggt ttcattgaag 1080

Sggggtggac agggatggta gatggatggt acggttatca ccatcaaaat gagcaggggt 1140 caggatatgc agccgacctg aagagcacac agaatgccat tgacgagatt actaacaaag 1200 taaattctgt tattgaaaag atgaatacac agttcacagc agtaggtaaa gagttcaacc 1260 acctggaaaa aagaatagag aatttaaata aaaaagttga tgatggtttc ctggacattt 1320 ggacttacaa tgccgaactg ttggttctat tggaaaatga aagaactttg gactaccacg 1380 attcaaatgt gaagaactta tatgaaaagg taagaagcca gctaaaaaac aatgccaagg 1440 aaattsgaaa cggctgcttt gaattttacc acaaatgcga taacacgtgc atggaaagtg 1500 tcaaaaatgg Gacttatgac tacccaaaat actcagagga agcaaaatta aacagagaag 1560 aaatagatgg ggtaaagctg gaatcaacaa ggatttacca gattttggcg atctattcaa 1620 ctgtcgccag ttcattggta ctggtagtct ccctgggggc aatcagtttc tggatgtgct 1680 ctaatgggtc tctacagtgt agaatatgta tttaacttta ggatttcaga agcat 1735

&Lt; 210 &gt; 13 &lt; 211 &gt; 1546 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 13 atcaatcact gagtgacatc gaagccatgg cgtctcaagg- caccaaacga tcatatgaac 60 aaatggagac tggtggggag cgccaggatg ccacagaaat cagagcatct gtcggaagaa 120 tgattggtgg aatcgggaga ttctacatcc aaatgtgcac tgaactcaaa ctcagtgatt 180 atgatggacg actaatccag aatagcataa caatagagag gatggtgctt tctgcttttg 240 atgagagaag aaataaatac ctagaagagc atcccagtgc tgggaaggac cctaagaaaa 300 caggaggacc catatataga agaatagacg gaaagtggat gagagaactc atcctttatg 360 acaaagaaga aataaggaga gtttggcgcc aagcaaacaa tggcgaagat gcaacagcag 420 gtcttactca tatcatgatt tggcattcca acctgaatga tgccacatat cagagaacaa 480 gagcgcttgt tcgcaccgga atggatccca gaatgtgctc tctaatgcaa ggttcaacac 540 ttcccagaag gtctggtgcc gcaggtgctg cggt £ aaagg Agttggaaca atagcaatgg 600 agttaatcag aatgatcaaa cgtggaatca atgaccgaaa tttctggagg ggtgaaaatg 660 gacgaaggac aagggttgct tatgaaagaa tgtgcaatat cctcaaagga aaatttcaaa 720 cagctgccca gagggcaats atggatcaag taagagaaag tcgaaaccca ggaaacgctg 780 agattgaaga cctcattttc ctggcacagt cagcactcat tctgaggg £ a tcagttgcac 840 ataaatcctg cctgcctgct tgtgtgtatg ggcttgcagt agcaagtggg catgactttg 900 aaagggaagg gtactcactg gtcgggatag acccattcaa attactccaa aacagccaag 960 tggtcagcct gatgagacca aatgaaaacc cagctcacaa gagtcaattg gtgtggatgg 1020 catgccactc tgctgcattt gaagatttaa gagtatcaag tttcataaga ggaaagaaag 1080 tgattccaag aggaaagctt tccacaagag gggtccagat tgcttcaaat gagaatgtgg 1140 -12- 159890 * sequence Listing .doc 201249996 aaaccatgga ctccaatacc ctggaactaa gaagcagata ctgggccata aggaccagga. 1200 gtggaggaaa taccaatcaa caaaaggcat ccgcaggcca gatcagtgtg cagcctacat 1260 tctcagtgca gcgaaatctc ccttttgaaa gagcaaccgt tatggcagca ttcagcggga 1320 acaatgaagg acggacatcc gacatgcgaa cagaagttat aagaatgatg gaaagtgcaa 1380 agccagaaga ttt £ tccttc caggggcggg gagtcttcga gctctcggac gaaaaggcaa 1440 cgaacccgat cgtgccttcc tttgacatga gtaatgaagg gtcttatttc ttcggagaca 1500 atgcagagga gtatgacagt tgaggaaaaa tacccttgtt tctact 1546 &lt;210&gt; 14 &lt;211&gt; 1410 &lt;212&gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 14 atgaatccaa accaaaagat aataaccatt ggttcggtct gtatgacgat tggaatggct 60 aacttaatat tacaaattgg aaacataatc tcaatatgga ttagccactc aattcaactt 120 gggaatcaaa atcagattga aacatgcaat caaagcgtca ttacttatga aaacaacact 180 tgggtaaatc agacatatgt taacatcagc aacaccaact ttgctgctgg acagtcagtg 240 gtttccgtga aattagcggg caattcctct ctctgccctg ttagtggatg ggctatatac 300 agtaaagaca acagtataag aatcggttcc aagggggatg tgtttgtcat aagggaacca 360 ttcatatcat gctccccctt ggaatgcaga accttcttct tgactcaagg ggccttgcta 420 aatgacaaac attccaatgg aaccattaaa gacaggagcc catatcgaac cctaatgagc 480 tgtcctattg gtgaagttcc ctctccatac aactcaagat ttgagtcagt cgcttggtca 540 gcaagtgctt gtcatgatgg catcaattgg ctaacaattg gaatttctgg cccagacaat 600 ggggcagtgg ctgtgttaaa gtacaacggc ataataacag acactatcaa gagttggaga 660 aacaatatat tgagaacaca agagtctgaa tgtgcatgtg taaatggttc Ttgctttact 720 gtaatgaccg atggaccaag tgatggacag gcctcataca agatcttcag aatagaaaag 780 ggaaagatag tcaaatcagt cgaaa tgaat gcccctaatt attactatga ggaatgctcc 840 tgttatcctg attctagtga aatcacatgt gtgtgcaggg ataactggca tggctcgaat 900 cgaccgtggg tgtctttcaa ccagaatctg gaatatcaga taggatacat atgcagtggg 960 attttcggag acaatccacg ccctaatgat aagacaggca gttgtggtcc agtatcgtct 1020 aatggagcaa atggagtaaa aggattttca ttcaaatacg gcaatggtgt ttggataggg 1080 agaactaaaa gcattagttc aagaaacggt tttgagatga tttgggatcc gaacggatgg 1140 actgggacag acaataactt ctcaataaag caagatatcg taggaataaa t £ agtggtca 1200 ggatatagcg ggagttttgt tcagcatcca gaactaacag ggctggattg tataagacct 1260 tgcttctggg ttgaactaat cagagggcga cccaaagaga acacaatctg gactagcggg 1320 agcagcatat ccttttgtgg tgtaaacagt gacactgtgg gttggtcttg gccagacggt 1380 gctgagttgc catttaccat tgacaagtaa 1410 ^ &gt; Λ &gt; &amp; 1 2 3 11 1i li Is &lt; 2 &lt; 2 &lt; 2 &lt; 2 15 1368

DNA H1N1 swine influenza virus • 13 · 159890 · Sequence Listing .doc 201249996 &lt; 400 &gt; 15 atgagtcttc taaccgaggt cgaaacgtac gttctttcta tcatcccgtc aggccccctc 60 aaagccgaga tcgcgcagag actggaaagt gtctttgcag gaaagaacac agatcttgag 120 gctctcatgg aatggctaaa gacaagacca atcttgtcac ctctgactaa gggaatttta 180 ggatttgtgt tcacgctcac cgtgcccagt gagcgaggac tgcagcgtag acgctttgtc 240 caaaatgccc taaatgggaa tggggacccg aacaacatgg acagagcagt taaactatac 300 aagaagctca aaagagaaat aacgttccat ggggccaagg aggtgtcact aagctattca 360 actggtgcac ttgccagttg catgggcctc atatacaaca ggatgggaac agtgaccaca 420 gaagctgctt ttggtctagt gtgtgccact tgtgaacaga ttgctgattc acagcatcgg 480 tctcacagac agatggctac taccaccaat ccactaatca ggcatgaaaa cagaatggtg 540 ctggctagca ctacggcaaa ggctatggaa cagatggctg gatcgagtga acagaaagga 600 gaagacgtcc aaaagctggc agaagagctg caaagcaaca ttggagtatt gagatctctt 660 ggagcaagtc Aaaagagtgg ggaaggaatt gcaaaggatg taatggaagt gctaaagcag 720 agctctatgg gaaattcagc tcttgtgaag aaatatctat gctcgaacca tttcagattc 780 ttt caatttg ttcttttatc ttatcagctc tccatttcat ggcttggaca aggcattatc 840 aaaaaagagg agagaaaaga aaaggtccaa acaaagagac aaacagagag gtatcaattt 900 gacacagtta ccaaaaagaa atccaggcca aagaaacaaa ggaagtactc tctgacaaca 960 agggtgacca caaggggctt tctgccggcg gaggccatgg aggttgctaa tcagactagg 1020 cagatggtac atgcaatgag aactattggg actcatccta gctccagtgc tggtctgaaa 1080 gatgaccttc ttgaaaattt gcaggcctac cagaagcgaa tgggagtgca gatgcagcga 1140 ttcaagtcct ctcgtcattg cagcaatatc attgggatct tgcacctgat attgtggatt 1200 actgatcgtc Tttttttcaa atgtatttat cgtcgcttta aatacggttt gaaaagaggg 1260 ccttctacgg aaggagtgcc tgagtccatg agggaagaat atcaacagga acagcagagt 1320 gctgtggatg ttgacgatgg tcattttgtc aacatagagc tagagtaa 1368

&lt;210&gt; 16 &lt;2]]&gt; 351 &lt;212&gt; DNA &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 16 aacataatgg actccaacac catgtcaagc tttcaggtag actgtttcct ttggcatatc 60 cgcaagcgat ttgcagacaa tggattgggt gatgccccat tccttgatcg gctccgccga 120 gatcaaaagt ccttaaaagg aagaggcaac acccttggcc tcgatatcga aacagccact 180 cttgttggga aacaaatcgt ggaatggatc ttgaaagagg aatccagcga gacacttaga 240 atgacaattg catctgtacc tacttcgcgc tacctttctg acatgaccct cgaggaaatg 300 tcacgagact ggttcatgct catgcctagg caaaagataa taggccctct ttgcgtgcga 360 ttggaccagg cgatcatgga aaagaacata gtactgaaag cgaacttcag tgtaatctct 420 aaccgattag agaccttgat actactaagg gctttcactg aggagggagc aatagttgga 480 gaaatttcac cattaccttc tcttccagga catacttatg aggatgtcaa aaatgcaatt 540 g £ ggtcctca tcggaggact Tgaatggaat ggtaacacgg ttcgagtctc tgaaaacta 600 cagagattcg cttggagaaa ctgtgatgag aatgggagac cttcactacc tccagagcag 660 • 14· 159890-sequence table doc 201249996 aaatgaaaag tggcgagagc aattgggaca gaaatttgag gaaataaggt ggttaattga 720 agaaatgcgg ca Cagattga aagcgacaga gaatagtttc gaacaaataa catttatgca 780 agccttacaa ctactgcttg aagtagaaca agagataaga gctttctcgt ttcagcttat 840 ttaactgata a 851 &lt;210&gt; 17 &lt;211&gt; 2293 &lt;212&gt; Wk &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 17 tatggagaga ataaaagaac tgagagatct aatgtcgcag tcccgcactc gcgagatact 60 cactaagacc actgtggacc atatggccat aatcaaaaag tacacatcag gaaggcaaga 120 gaagaacccc gcactcagaa tgaagtggat gatggcaatg agatacccaa ttacagcaga 180 caagagaata atggacatga ttccagagag gaatgaacaa ggacaaaccc tctggagcaa 240 aacaaacgat gctggatcag accgagtgat ggtatcacct ctggccgtaa catggtggaa 300 taggaatggc ccaacaacaa gtacagttca ttaccctaag gtatataaaa cttatttcga 360 aaaggtcgaa aggttgaaac atggtacctt cggccctgtc cacttcagaa atcaagttaa 420 aataaggagg agagttgata caaaccctgg ccatgcagat ctcagtgcca Aggaggcaca 480 ggatgtgatt atggaagttg ttttcccaaa tgaagtgggg gcaagaatac tgacatcaga 540 gtcacagctg gcaataacaa aagagaagaa agaagagctc caggattgta aaattgctcc 600 cttgatggtg gcgtacatgc tagaaagaga attggtccgt aaaacaaggt ttctcccagt 660 agccggcgga acaggcagtg tttatattga agtgttgcac ttaacccaag ggacgtgctg 720 ggagcagatg tacactccag gaggagaagt gagaaatgat gatgttgacc aaagtttgat 780 tatcgctgct agaaacatag taagaagagc agcagtgtca gcagacccat tagcatctct 840 cttggaaatg tgccacagca cacagattgg aggagtaagg atggtggaca tccttagaca 900 gaatccaact gaggaacaag ccgtagacat atgcaaggca gcaatagggt tgaggattag 960 ctcatctttc agttttggtg ggttcacttt caaaaggaca agcggatcat cagtcaagaa 1020 agaagaagaa gtgctaacgg gcaacctcca aacactgaaa ataagagtac atgaagggta 1080 tgaagaattc acaatggttg ggagaagagc aacagctatt ctcagaaagg caaccaggag 1140 attgatccag ttgatagtaa gcgggagaga cgagcagtca attgctgagg caataattgt 1200 ggccatggta ttctcacaag aggattgcat gatcaaggca gttaggggcg atctgaactt 1260 tgtcaatagg gcaaaccagc gactgaaccc catgcaccaa ctcttgaggc atttccaaaa 1320 agatgcaaaa gtgcttttcc agaactgggg aattgaatcc atcgacaatg tgatgggaat 1380 gatcggaata ctgcccgaca tgaccccaag cacggagatg tcgctgagag ggataagagt 1440 cagcaaaatg ggagtagatg aatactccag cacggagaga gtggtagtga gtattgaccg 1500 atttttaagg gttagagatc aaagagggaa cgtactattg tctcccgaag aagtcagtga 1560 aacgcaagga actgagaagt tgacaataac ttattcgtca tcaatgatgt gggagatcaa 1620 tggccctgag tcagtgctag tcaacactta tcaatggata atcaggaact gggaaattgt 1680 gaaaattcaa tggtcacaag atcccacaat gttatacaac aaaatggaat ttgaaccatt 1740 tcagtctctt gtccctaagg caaccagaag ccggtacagt ggattcgtaa ggacactgtt 1800 • 15- 159890- sequence table .doc 201249996 ccagcaaatg cgggatgtgc ttgggacatt tgacactgtc caaataataa aacttctccc 1860 ctttgctgct gctccaccag aacagagtag gatgcaattt tcctcattga ctgtgaatgt 1920 gagaggatca £ ggttgag £ a tactggtaag aggcaattct ccagtattca attacaacaa 1980 ggcaaccaaa cgacttacag ttcttggaaa ggatgcaggt gcattgactg aagatccaga 2040 tgaaggcaca tctggggtgg agtctgctgt cctgagagga tttctcattt tgggcaaaga 2100 agacaagaga tatggcccag cattaagcat caatgaactg agcaatcttg caaaaggaga 2160 gaaagctaat gtgctaattg ggcaagggga cgtagtgttg gtaatgaaac gaaaacggga 2220 ctctagcata cttactgaca Gccagacagc gaccaaaaga attcggatgg ccatcaatta 2280 gtgtcgaatt gtt 2293

&lt;210&gt; 18 &lt;211&gt; 2303 &lt;2]2&gt; DNA &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 18 ttgaatggat gtcaatccga ctctactttt cctaaaaatt ccagcgcaaa atgccataag 60 caccacattc ccttatactg gagatcctcc atacagccat ggaacaggaa caggatacac 120 catggacaca gtaaacagaa cacaccaata ctcagaaaag ggaaagtgga cgacaaacac 180 agagactggt gcaccccagc tcaacccgat tgatggacca ctacctgagg ataatgaacc 240 aagtgggtat gcacaaacag actgtgttct agaggctatg gctttccttg aagaatccca 300 cccaggaata tttgagaatt catgccttga aacaatggaa gttgttcaac aaacaagggt 360 agataaacta actcaaggtc gccagactta tgattggaca ttaaacagaa atcaaccggc 420 agcaactgca ttggccaaca ccatagaagt ctttagatcg aatggcctaa cagctaatga 480 gtcaggaagg ctaatagatt tcttaaagga tgtaatggaa tcaatgaaca aagaggaaat 540 agagataaca acccactttc aaagaaaaag gagagtaaga gacaacatga Ccaagaagat 600 ggtcacgcaa agaacaatag ggaagaaaaa acaaagactg aataagagag gctatctaat 660 aagagcactg acattaaata cgatgaccaa agatgcagag agaggcaagt taaaaagaag 720 ggctatcgca acacctggga tgcagattag aggtttcgta tactttgttg aaactttagc 780 taggagcatt tgcgaaaagc ttgaacagtc tgggctccca gtag £ gggca atgaaaagaa 840 ggccaaactg gcaaatgttg tgagaaagat gatgactaat cagagatttc 900 tttcacaatc actggggaca acactaagtg gaatgaaaat caaaatcctc gaatgttcct 960 ggcgatgatt acatatatca ccagaaatca acccgagtgg ttcagaaaca tcctgagcat 1020 ggcacccata atgttctcaa acaaaatggc aagactaggg aaagggtaca tgttcgagag 1080 taaaagaatg aagattcgaa cacaaatacc agcagaaatg ctagcaagca ctgacctgaa 1140 gtacttcaat gaatcaacaa agaagaaaat tgagaaaata tcacaagaca aggcctcttc taatagatgg 1200 cacagcatca ctgagtcctg ggatgatgat gggcatgttc aacatgctaa gtacggtctt 1260 gggagtctcg atactgaatc ttggacaaaa ^ aaatacacc aagacaatat actggtggga 1320 tgggctccaa tcatccgacg attttgctct catagtgaat gcaccaaacc atgagggaat 1380 acaagcagga gtggacagat tctacaggac ctgcaagtta gtgggaatca acatgagcaa 1440 aaagaagtcc tatataaata agacagggac atttgaattc acaagctttt tttatcgcta 1500 -16 · 159890- d〇c 201 249 996 sEQUENCE LISTING Tggatttgtg gctaatttta gcatggagct acccagcttt ggagtgtctg gagtaaatga 1560 atcagctgac atgagtattg gagt aacagt gataaagaac aacatgataa acaatgacct 1620 tggacctgca acggcccaga tggctcttca attgttcatc aaagactaca gatacacata 1680 taggtgccat aggggagaca cacaaattca gacgagaaga tcatttgagt taaagaagct 1740 gtgggatcaa acccaatcaa aggtagggct attagtatca gatggaggac caaacttata 1800 caatatacgg aatcttcaca ttcctgaagt ctgcttaaaa tgggagctaa tggatgatga 1860 ttatcgggga agactttgta atcccctgaa tccctttgtc agtcataaag agattgattc 1920 tgtaaacaat gctgtggtaa tgccagccca tggtccagcc aaaagcatgg aatatgatgc 1980 cgttgcaact acacattcct ggattcccaa gaggaatcgt tctattctca acacaagcca 2040 aaggggaatt cttgaggatg aacagatgta ccagaagtgc tgcaatctat tcgagaaatt 2100 tttccctagc agttcatata ggagaccggt tggaatttct agcatgglgg aggccatggt 2160 gtctagggcc cggattgatg ccagggtcga cttcgagtct ggacggatca agaaagaaga 2220 gttctctgag atcatgaaga tctgttccac cattgaagaa ctcagacggc aaaaataatg 2280 aatttaactt gtccttcatg aaa 2303

&Lt; 210 &gt; 19 &lt; 211 &gt; 2172 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 19 tccaaaatgg aagactttgt gcgacaatgc ttcaatccaa tgatcgtcga gcttgcggaa 60 aaggcaatga aagaatatgg ggaagatccg aaaatcgaaa ctaacaagtt tgctgcaata 120 tgcacacatt tggaagtttg tttcatgtat tcggatttcc atttcatcga cgaacggggt 180 gaatcaataa ttgtagaatc tggtgacccg aatgcactat tgaagcaccg atttgagata 240 attgaaggaa gagaccgaat catggcctgg acagtggtga acagtatatg taacacaaca 300 gg £ gta £ aga agcctaaatt tcttcctgat acaaagagaa ccggttcatt 360 gaaattggag taacacggag ggaagtccac atatattacc tagagaaagc caacaaaata 420 aaatctgaga agacacacat tcacatcttt tcattcactg gagaggagat ggccaccaaa 480 gcggactaca cccttgacga agagagcagg gcaagaatca aaactaggct tttcactata 540 agacaagaaa tggccagtag gagtctatgg ttgtatgatt Gattcctttc gtcagtccga aagaggcgaa 600 gagacaattg aagaaaaatt tgagattaca ggaactatgc gcaagcttgc cgaccaaagt 660 ctcccaccga acttctccag ccttgaaaac tttagagcct atgtagatgg attcgagccg 720 aacggctgca ttgagggcaa gctttcccaa atgtcaaaag aagtgaacgc caaaattgaa 780 ccattcttga ggacgacacc acgccccctc agattgcctg atgggcctct ttgccatcag 840 cggtcaaagt tcctgctgat aaattaagta ggatgctctg gtagactttg atgactgcaa agatgttgga ttgaagaccc gagtcacgag 900 ggggagggaa taccactata tgatgcaatc aaatgcatga agacattctt tggctggaaa 960 gagcctaaca tagtcaaacc.acatgagaaa ggcataaatc ccaattacct catggcttgg 1020 aagcaggtgc tagcagagct acaggacatt gaaaatgaag agaagatccc aaggacaaag 1080 aacatgaaga gaacaagcca attgaagtgg gcactcggtg aaaatatggc accagaaaaa 1140 gaccttaaac agtatgacag tgatgagcca 1200 • 17- 159890 · sequence Listing .doc 201249996 gagcccagat ctctagcaag ctgggtccaa aatgaattca ataaggcatg tgaattgact gattcaagct ggatagaact tgatgaaata ggagaagatg ttgccccgat tgaacatatc gcaagcatga ggaggaacta ttttacagca gaagtgtccc actgcagggc tactgaatac ataatgaagg gagt £ tacat aaatacggcc ttgctcaatg catcctgt £ c agccatggat gactttcagc tgatcccaat gataagcaaa tgtaggacca aagaaggaag acggaaaaca aacctgtatg ggttcattat aaaaggaagg tctcatttga Gaaatgatac tgatgtggtg aactttgtaa gtatggagtt ctcactcact gacccgagac tggagccac a caaatgggaa aaatactgtg ttcttgaaat aggagacatg ctcttgagga ctgcgatagg ccaagtgtcg aggcccatgt tcctatatgt gagaaccaat ggaacctcca agatcaagat gaaatggggc atggaactga ggcgctgcct tcttcagtct cttcagcaga ttgagagcat gattgaggcc gagtcttctg tcaaagagaa agacatgacc aaggaattct ttgaaaacaa atcggaaaca tggccaatcg gagagtcacc caggggagtg gaggaaggct ctattgggaa agtgtgcagg accttactgg caaaatctgt attcaacagt ctatatgcgt ctccacaact tgaggggttt tcggctgaat cgagaaaatt gcttctcatt gttcaggcac ttagggacaa cctggaacct ggaaccttcg atcttggggg gctatatgaa gcaatcgagg agtgcctgat taatgatccc Tgggttttgc ttaatgcatc ttggttcaac tccttcctca cacatgcact gaagtagttg tggcaatgct ac

&Lt; 210 &gt; 20 &lt; 211> 1735 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 20 aaaagcaaca aaagatgaag gcaatactag tagttctgct atatacattt gcaaccgcaa atgcagacac attatgtata ggttatcatg cgaacaattc aacagacact gtagacacag tactagaaaa gaatgtaaca gtaacacact ctgttaacct tctagaagac aagcataacg ggaaactatg caaactaaga ggggtagccc cattgcattt gggtaaatgt aacattgctg gctggatcct gggaaatcca gagtgtgaat cactctccac agcaagctca tggtcctaca ttgtggaaac atctagttca gacaatggaa cgtgttaccc aggagatttc atcgattatg aggagctaag agagcaattg tacagcaaga agttcaagcc agctcagtgt catcatttga aaggtttgag atattcccca agacaagttc atggcccaat catgactcga acaaaggtgt aacggcagca tgtcctcatg ctggagcaaa aagcttctac aaaaatttaa tatggctagt taaaaaagga aattcatacc caaagctcag caaatcctac attaatgata aagggaaaga agtcctcgtg ctatggggca ttcaccatcc atctactagt gctgaccaac aaagtctcta tcagaatgca gatgcatatg tttttgtggg gtcatcaaga Ggaaatagca ataagaccca aagtgaggga tcaagaaggg agaatgaact attactggac actagtagag ccgggagaca aaataacatt cgaagcaact ggaaatctag tggtac cgag atatgcattc gcaatggaaa gaaatgctgg atctggtatt atcatttcag atacaccagt ccacgattgc aatacaactt gtcagacacc caagggtgct ataaacacca gcctcccatt tcagaatata catccgatca caatt £ gaaa atgtccaaaa tatgtaaaaa gcacaaaatt gagactggcc acaggattga 159890- Sequence Listing .doc 18- 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2172 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 201249996 ggaatgtccc gtctattcaa tctagaggcc tatttggggc cattgccggt ttcattgaag 1080 gggggtggac agggatggta gatggaiggt acggttatca ccatcaaaat gagcaggggt 1140 caggatatgc agccgacctg aagagcacac agaatgccat tgacgagatt actaacaaag 1200 taaattctgt tattgaaaag atgaatacac agttcacagc agtaggtaaa gagttcaacc 1260 acctggaaaa aagaatagag aatttaaata aaaaagttga tgatggtttc ctggacattt 1320 ggacttacaa tgccgaactg ttggttctat tggaaaat£a aagaactttg gactaccacg 1380 attcaaatgt gaagaactta tatgaaaagg taagaagcca gctaaaaaac aatgccaagg 1440 aaattggaaa cggctgcttt gaattttacc acaaatgcga taacacgtgc atggaaagtg 1500 tcaaaaatgg gactta Tgac tacccaaaat actcagagga agcaaaatta aacagagaag 1560 aaatagatgg ggtaaagctg gaatcaacaa ggatttacca gattttggcg atctattcaa 1620 ctgtcgccag ttcattggta ctggtagtct ccctgggggc aatcagtttc tggatgtgct 1680 ctaatgggtc tctacagtgt agaatatgta tttaacttta ggatttcaga agcat 1735

&Lt; 210 &gt; 21 &lt; 211 &gt; 1546 &lt; 212> DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 21 atcaatcact gagtgacatc gaagccatgg cgtctcaagg caccaaacga tcatatgaac 60 aaatggagac tggtggggag cgccaggatg ccacagaaat cagagcatct gtcggaagaa 120 tgattggtgg aatcgggaga ttctacatcc aaatgtgcac tgaactcaaa ctcagtgatt 180 atgatggacg actaatccag aatagcataa caatagagag gatggtgctt tctgcttttg 240 atgagagaag aaataaatac ctagaagagc atcccagtgc tgggaaggac cctaagaaaa 300 caggaggacc catatataga agaatagacg gaaagtggat gagagaactc atcctttatg 360 acaaagaaga aataaggaga gtttggcgcc aagcaaacaa tggcgaagat gcaacagcag 420 gtcttactca tatcatgatt tggcattcca acctgaatga tgccacatat cagagaacaa 480 gagcgcttgt tcgcaccgga atggatccca gaatgtgctc tctaatgcaa ggttcaacac 540 ttcccagaag gtctggtgcc gcaggtgctg cggtgaaagg agttggaaca atagcaatgg 600 Agttaatcag aatgatcaaa cgtggaatca atgaccgaaa tttctggagg ggtgaaaatg 660 gacgaaggac aagggttgct tatgaaagaa tgtgcaatat cctcaaagga aaatttcaaa 720 cagctgccca gagggcaatg atggatcaag taagagaaag tcgaaaccca gga aacgctg 780 agattgaaga cctcattttc ctggcacggt cagcactcat tctgagggga 840 tcagttgcac ctccaatacc ctggaactaa gaagcagata ataaatcctg cctgcctgct tgtgtgtatg ggcttgcagt agcaagtggg catgactttg 900 aaagggaagg gtactcactg gtcgggatag acccattcaa attactccaa aacagccaag 960 tggtcagcct gatgagacca aatgaaaacc cagctcacaa gagtcaattg gtgtggatgg 1020 catgccactc tgctgcattt gaagatttaa gagtatcaag tttcataaga ggaaagaaag 1080 tgattccaag aggaaagctt tccacaagag gggtccagat tgcttcaaat gagaatgtgg 1140 aaaccatgga ctgggccata aggaccagga 1200 gtggaggaaa taccaatcaa caaaaggcat ccgcaggcca gatcagtgtg cagcctacat 1260 tctcagtgca gcgaaatctc ccttttgaaa gagcaaccgt tatggcagca ttcagcggga 1320 • 19 · 159890- sequence table .doc 201249996 acaatgaagg acggacatcc gacatgcgaa cagaagttat aagaatgatg gaaagtgcaa 1380 agccagaaga tttgtccttc caggggcggg gagtcttcga gctctcggac gaaaaggcaa 1440 cgaacccgat cgtgccttcc tttgacatga gtaatgaagg gtcttatttc ttcggagaca 1500 atgcagagga gtatgacagt tgaggaaaaa tacccttgtt Tctact 1546 &lt;210&gt; 22 &lt;211&gt; 1414 &lt;212&gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 22 tgatgaatcc aaaccaaaag ataataacca ttggttcggt ctgtatgaca attggaatgg 60 ctaacttaat gttacaaatt ggaaacataa tctcaatatg gattagccac tcaattcaac 120 ttgggaatca aaatcagatt gaaacatgca atcaaagcgt cattacttat gaaaacaaca 180 cttgggtaaa tcagacatat gttaacatca gcaacaccaa ctttgctgot ggacagtcag 240 tggtttccgt gaaattagcg ggcaattcct ctctctgccc tgttagtgga tgggctatat 300 acagtaaaga caacagtgta agaatcggtt ccaaggggga tgtgtttgtc ataagggaac 360 cattcatatc atgctccccc ttggaatgca gaaccttctt cttgactcaa ggggccttgc 420 taaatgacaa acattccaat ggaaccatta aagacaggag cccatatcga accctaatga 480 gctgtcctat tggtgaagtt ccctctccat acaactcaag atttgagtca gtcgcttggt 540 cagcaagtgc ttgtcatgat ggcatcaatt ggctaacaat tggaatttct ggcccagaca 600 atggggcagt ggctgtgtta aagtacaacg gcataataac agacactatc aagagttgga 660 gaaacaatat attgagaaca c.aagagtctg aatgtgcatg tgtaaatggt Tcttgcttta 720 ctgtaatgac cgatggacca agtaatgaac aggcctcata caagatcttc agaatagaaa 780 agggaaagat agtcaaatca gtcgaaat ga atgcccctaa ttatcactat gaggaatgct 840 cctgttatcc tgattctagt gaaatcacat gtgtgtgcag ggataactgg catggctcga 900 atcgaccgtg ggtgtctttc aaccagaatc tggaatatca gataggatac atatgcagtg 960 ggattttcgg agacaatcca cgccctaatg ataagacagg cagttgtggt ccagtatcgt 1020 ctaatggagc aaatggagta aaaggatttt cattcaaata cggcaatggt gtttggatag 1080 ggagaactaa aagcattagt tcaagaaacg gttttgagat gatttgggat ccgaacggat 1140 ggactgggac agacaataac ttctcaataa agcaagatat cgtaggaata aatgagtggt 1200 caggatatag cgggagtttt gttcagcatc Cagaactaac agggctggat tgtataagac 1260 cttgcttctg ggttgaacta atcagagggc gacccaaaga gaacacaatc tggactagcg 1320 ggagcagcat atccttttgt ggtgtaaaca gtgacactgt gggttggtct tggccagacg 1380 gtgctgagtt gccatttacc attgacaagt aaca 1414

&lt;210&gt; 23 &lt;211&gt; 1368 &lt;212&gt; DNA &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 23 atgagtcttc taaccgaggt cgaaacgtac gttctttcta tcatcccgtc aggccccctc 60 aaagccgaga tcgcgcagag actggaaagt gtctttgcag gaaagaacac agatcttgag 120 •20· 159890· list .doc 201249996 gctctcatgg aatggctaaa gacaagacca atcttgtcac ctctgactaa gggaatttta 180 ggatttgtgt tcacgctcac cgtgcccagt gagcgaggac tgcagcgtag acgctttgtc 240 caaaatgccc taaatgggaa tggggacccg aacaacatgg atagagcagt taaactatac. 300 aagaagctca aaagagaaat aacgttccat ggggccaagg aggtgtcact aagctattca 360 actggtgcac ttgccagttg catgggcctc atatacaaca ggatgggaac agtgaccaca 420 gaagctgctt ttggtctagt gtgtgccact tgtgaacaga ttgctgattc acagcatcgg 480 tctcacagac agatggctac taccaccaat Ccactaatca ggcatgaaaa cagaatggtg 540 ctggctagca ctacggcaaa ggctatggaa cagatggctg gatcgagtga acagaaagga 600 gaagacgtcc aaaagctggc agaagagctg caaagcaaca ttggagtatt gagatctctt 660 ggagcaagtc aaaagagtgg ggaaggaatt £caaag£atg taatggaagt gctaaagcag 720 agctctatgg ga aattcagc tcttgtgaag aaatatctat gctcgaacca tttcagattc 780 tttcaatttg ttcttttatc ttatcagctc tccatttcat ggcttggaca aggcattatc 840 aaaaaagagg agagaaaaga aaaggtccaa acaaagagac aaacagagag gtatcaattt 900 gacacagtta ccaaaaagaa atccaggcca aagaaacaaa ggaagtactc tctgacaaca 960 agggtgacca caaggggctt tctgccggcg gaggccatgg aggttgctaa tcagactagg 1020 cagatggtac atgcaatgag aactattggg actcatccta gctccagtgc tggtctgaaa 1080 gatgaccttc ttgaaaattt gcaggcctac cagaagcgaa tgggagtgca gatgcagcga 1140 ttcaagtcct ctcgtcattg Cagcaatatc attgggatct tgcacctgat attgtggatt 1200 actgatcgtc tttttttcaa atgtatttat cgtcgcttta aatacggttt gaaaagaggg 1260 ccttctacgg aaggagtgcc tgagtccatg agggaagaat atcaacagga acagcagagt 1320 gctgtggatg ttgacgatgg tcattttgtc aacatagagc tagagtaa 1368

&Lt; 210 &gt; 24 &lt; 211 &gt; 851 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 24 aacataatgg actccaacac catgtcaagc tttcaggtag actgtttcct ttggcatatc 60 cgcaagcgat ttgcagacaa tggattgggt gatgccccat tccttgatcg gctccgccga 120 gatcaaaagt ccttaaaagg aagaggcaac acccttggcc tcgatatcga aacagccact 180 cttgttggga aacaaatcgt ggaatggatc ttgaaagagg aatccagcga gacacttaga 240 atgacaattg catctgtacc tacttcgcgc tacctttctg acatgaccct cgaggaaatg 300 tcacgagact ggttcatgct catgcctagg caaaagataa taggccctct ttgcgtgcga 360 ttggaccagg cgatcatgga aaagaacata gtactgaaag cgaacttcag tgtaatcttt 420 aaccgattag agaccttgat actactaagg gctttcactg aggagggagc aatagttgga 480 gaaatttcac cattaccttc tcttccagga catacttatg aggatgtcaa aaatgcaatt 540 g £ ggtcctca tcggaggact tgaatggaat ggtaacacgg ttcgagtctc Tgaaaatata 600 cagagattcg cttggagaaa ctgtgatgag aatgggagac cttcactacc tccagagcag 660 aaatgaaaag tggcgagagc aattgggaca gaaatttgag gaaataaggt ggttaattga 720 agaaatgcgg cacagattga aagcgacaga gaatagtttc gaacaaataa ca Tttatgca 780 21 · 159890 - Sequence Listing.doc 201249996 agccttacaa ctactgcttg aagtagaaca agagataaga gctttctcgt ttcagcttat 840 ttaactgata a 851 &lt;210&gt; 25 &lt;211&gt; 2293 &lt;212&gt; DNA &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 25 tatggagaga ataaaagaac tgagagatct aatgtcgcag tcccgcactc gcgagatact 60 cactaagacc actgtggacc atatggccat aatcaaaaag tacacatcag gaaggcaaga 120 gaagaacccc gcactcagaa tgaagtggat gatggcaatg agatacccaa ttacagcaga 180 caagagaata atggacatga ttccagagag gaatgaacaa ggacaaaccc tctggagcaa 240 aacaaacgat gctggatcag accgagtgat ggtatcacct ctggccgtaa catggtggaa 300 taggaatggc ccaacaacaa gtacagttca ttaccctaag gtatataaaa cttatttcga 360 aaaggtcgaa aggttgaaac atggtacctt cggccctgtc cacttcagaa atcaagttaa 420 Aataaggagg agagttgata caaaccctgg ccatgcagat ctcagtgcca aggaggcaca 480 ggatgtgatt atggaagttg ttttcccaaa tgaagtgggg gcaagaatac tgacatcaga 540 gtcacagctg gcaataacaa aagagaagaa agaagagctc caggattgta aaattgctcc 600 cttgatggtg gcgtacatgc tagaaagaga attggtccgt aaaacaaggt ttctc ccagt 660 agccggcgga acaggcagtg tttatattga agtgttgcac ttaacccaag ggacgtgctg. 720 ggagcagatg tacactccag gaggagaagt gagaaatgat gatgttgacc aaagtttgat 780 tatcgctgct agaaacatag taagaagagc agcagtgtca gcagacccat tagcatctct 840 cttggaaatg tgccacagca cacagattgg aggagtaagg atggtggaca tccttagaca 900 gaatccaact gaggaacaag ccgtagacat atgcaaggca gcaatagggt tgaggattag 960 ctcatctttc agttttggtg ggttcacttt caaaaggaca agcggatcat cagtcaagaa 1020 agaagaagaa gtgctaacg £ gcaacctcca aacactgaaa ataagagtac atgaagggta 1080 tgaagaattc acaatggttg ggagaagagc aacagctatt ctcagaaagg caaccaggag 1140 attgatccag ttgatagtaa gcgggagaga cgagcagtca attgctgagg caataattgt 1200 ggccatggta ttctcacaag aggattgcat gatcaaggca gttaggggcg atctgaactt 1260 tgtcaatagg gcaaaccagc gactgaaccc catgcaccaa ctcttgaggc atttccaaaa 1320 agatgcaaaa gtgcttttcc agaactgggg aattgaatcc atcgacaatg tgatgggaat 1380 gatcggaata ctgcccgaca tgaccccaag cacggagatg tcgctgagag ggataagagt 1440 cagcaaaatg ggagtagatg aatactccag cacggagaga gtggtagtga Gtattgaccg 150 0 atttttaagg gttagagatc aaagagggaa cgtactattg tctcccgaag aagtcagtga 1560 aacgcaagga actgagaagt tgacaataac ttattcgtca tcaatgatgt gggagatcaa 1620 tggccctgag tcagtgctag tcaacactta tcaatggata atcaggaact gggaaattgt 1680 gaaaattcaa tggtcacaag atcccacaat gttatacaac aaaatggaat ttgaaccatt 1740 tcagtctctt gtccctaagg caaccagaag ccggtacagt ggattcgtaa ggacactgtt 1800 ccagcaaatg cgggatgtgc ttgggacatt tgacactgtc caaataataa aacttctccc I860 ctttgctgct gctccaccag aacagagtag gatgcaattt tcctcattga ctgtgaatgt 1920 • 22 · 159890 · sequence Listing d〇c 201249996 gagaggatca gggttgagga tactggtaa £ aggcaattct ccagtattca attacaacaa 1980 ggcaaccaaa cgacttacag ttcttggaaa ggatgcaggt gcattgactg aagatccaga 2040 tgaaggcaca tctggggtgg agtctgctgt cctgagagga tttctcattt tgggcaaaga 2100 agacaagaga tatggcccag cattaagcat caatgaactg agcaatcttg caaaaggaga 2160 gaaagctaat gtgctaattg ggcaagggga cgtagtgttg gtaatgaaac gaaaacggga 2220 ctctagcata cttactgaca gccagacagc Gaccaaaaga attcggatgg ccatcaat.ta 2280 gtgtcgaatt gtt 2293

&Lt; 210 &gt; 26 &lt; 211 &gt; 2303 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 26 ttgaatggat gtcaatccga ctctactttt cctaaaaatt ccagcgcaaa atgccataag 60 caccacattc ccttatactg gagatcctcc atacagccat ggaacaggaa caggatacac 120 catggacaca gtaaacagaa cacaccaata ctcagaaaag ggaaagtgga cgacaaacac 180 agagactggt gcaccccagc tcaacccgat tgatggacca ctacctgagg ataatgaacc 240 aagtgggtat gcacaaacag actgtgttct agaggctatg gctttccttg aagaatccca 300 cccaggaata tttgagaatt catgccttga aacaatggaa gttgttcaac aaacaagggt 360 agataaacta actcaaggtc gccagactta tgattggaca ttaaacagaa atcaaccggc 420 agcaactgca ttggccaaca ccatagaagt ctttagatcg aatggcctaa cagctaatga 480 gtcaggaagg ctaatagatt tcttaaagga tgtaatggaa tcaatgaaca aagaggaaat 540 agagataaca acccactttc aaagaaaaag gagagtaaga gacaacatga ccaagaagat 600 Ggtcacgcaa agaacaatag ggaagaaaaa acaaagactg aataagagag gctatctaat 660 aagagcactg acattaaata cgatgaccaa agatgcagag agaggcaagt taaaaagaag 720 ggctatcgca acacctggga tgcagattag aggtttcgta tactttgttg aa actttagc 780 taggagcatt tgcgaaaagc tt £ aacagtc tgggctccca gtagggggca atgaaaagaa 840 ggccaaactg gcaaatgttg tgagaaagat gatgactaat cagagatttc 900 tttcacaatc actggggaca acactaagtg gaatgaaaat caaaatcctc gaatgttcct 960 ggcgatgatt acatatatca ccagaaatca acccgagtgg ttcagaaaca tcctgagcat 1020 ggcacccata atgttctcaa acaaaatggc aagactaggg aaagggtaca tgttcgagag 1080 taaaa £ aatg aagattcgaa cacaaatacc agcagaaatg ctagcaagca ttgacctgaa 1140 gtacttcaat gaatcaacaa tcacaagaca agaagaaaat tgagaaaata aggcctcttc taatagatgg 1200 cacagcatca ctgagtcctg ggatgatgat gggcatgttc aacatgctaa gtacggtctt 1260 gggagtctcg atactgaatc ttggacaaaa gaaatacacc aagacaatat actggtggga 1320 tgggctccaa tcatccgacg attttgctct catagtgaat gcaccaaacc atgagggaat 1380 acaagcagga gtggacagat tctacaggac ctgcaagtta gtgggaatca acatgagcaa 1440 aaagaagtcc tatataaata agacagggac atttgaattc acaagctttt tttatcgcta 1500 tggatttgtg gctaatttta gcatggagct acccagcttt ggagtgtctg gagtaaatga 1560 atcagctgac atgagtattg gagtaacagt Gataaagaac aacatgataa acaatgacct 1620 of the sequence listing -23-159890- .doc 201249996 tggacctgca acggcccaga tggctcttca attgttcatc aaagactaca gatacacata 1680 taggtgccat aggggagaca cacaaattca gacgagaaga tcatttgagt taaagaagct 1740 gtgggatcaa acccaatcaa aggtagggct attagtatca gatggaggac caaacttata 1800 caatatacgg aatcttcaca ttcctgaagt ctgcttaaaa tgggagctaa tggatgatga 1860 ttatcgggga agactttgta atcccctgaa tccctttgtc agtcataaag agattgattc 1920 tgtaaacaat gctgtggtaa tgccagccca tggtccagcc aaaagcatgg aatatgatgc 1980 cgttgcaact acacattcct ggattcccaa gaggaatcgt tctattctca acacaagcca 2040 aaggggaatt cttgaggatg aacagatgta ccagaagtgc tgcaatctat tcgagaaatt 2100 tttccctagc agttcatata ggagaccggt tggaatttct agcatggtgg aggccatggt 2160 gtctagggcc cggattgatg ccagggtcga cttcgagtct ggacggatca agaaagaaga 2220 gttctctgag atcatgaaga tctgttccac cattgaagaa ctcagacggc aaaaataatg 2280 aatttaactt gtccttcatg aaa 2303

&Lt; 210 &gt; 27 &lt; 211 &gt; 2172 &lt; 212 &gt; DNA &lt; 213 &gt; H1N1 swine influenza virus · &lt; 400 &gt; 27 tccaaaatgg aagactttgt gcgacaatgc ttcaatccaa tgatcgtcga gcttgcggaa 60 aaggcaatga aagaatatgg ggaagatccg aaaatcgaaa ctaacaagtt tgctgcaata 120 tgcacacatt tggaagtttg tttcatgtat tcggatttcc atttcatcga cgaacggggt 180 gaatcaataa ttgtagaatc tggtgacccg aatgcactat tgaagcaccg atttgagata 240 attgaaggaa gagaccgaat catggcctgg acagtggtga acagtatatg taacacaaca 300 ggggtagaga agcctaaatt tcttcctgat ttgtatgatt acaaagagaa ccggttcatt 360 gaaattggag taacacggag ggaagtccac atatattacc tagagaaagc caacaaaata 420 aaatctgaga agacacacat tcacatcttt tcattcactg gagaggagat ggccaccaaa 480 gcggactaca cccttgacga agagagcagg gcaagaatca aaactaggct tttcactata 540 agacaagaaa tggccagtag gagtctatgg gattcctttc gtcagtccga aagaggcgaa 600 gagacaattg aagaaaaatt tgagattaca g£aactatgc gcaagcttgc cgaccaaagt 660 ctcccaccga acttctccag ccttgaaaac tttagagcct atgtagatgg attcgagccg 720 aacggctgca ttgagggcaa gctttcccaa atgtcaaaag aagtgaacg c caaaattgaa 780 ccattcttga ggacgacacc acgccccctc agattgcctg atgggcctct ttgccatcag 840 cggtcaaagt tcctgctgat aaattaagta ggatgctctg gtagactttg atgactgcaa agatgttgga ttgaagaccc gagtcacgag 900 ggggagggaa taccactata tgatgcaatc aaatgcatga agacattctt tggctggaaa 960 gagcctaaca tagtcaaacc acatgagaaa ggcataaatc ccaattacct catggcttgg 1020 aagcaggtgc tagcagagct acaggacatt gaaaatgaag agaagatccc aaggacaaag 1080 aacatgaaga gaacaagcca attgaagtgg gcactcggtg aaaatatggc accagaaaaa 1140 gaccttaaac agtatgacag tgatgagcca 1200 gagcccagat ctctagcaag ctgggtccaa aatgaattca ataaggcatg tgaattgact 1260 gattcaagct ggatagaact tgatgaaata ggagaagatg ttgccccgat tgaacatatc 1320 -24- 159890 · sequence Listing .doc 201249996 gcaagcatga ggaggaacta ttttacagca gaagtgtccc actgcagggc tactgaatac 1380 ataatgaagg gagtgtacat aaatacggcc ttgctcaatg catcctgtgc agccatggat 1440 gactttcagc tgatcccaat gataagcaaa tgtaggacca aagaaggaag acggaaaaca 1500 aacctgtatg ggttcattat aaaaggaagg Tctcatttga gaaatgatac tgatgtggtg 1560 aactttgtaa gtatggagtt ctca ctcact gacccgagac tggagccaca caaatgg £ aa 1620 aaatactgtg ttcttgaaat aggagacatg ctcttgagga ctgcgatagg ccaagtgtcg 1680 aggcccatgt tcctatatgt gagaaccaat ggaacctaca agatcaagat gaaatggggc 1740 atggaactga ggcgctgcct tcttcagtct cttcagcaga ttgagagcat gattgaggcc 1800 gagtcttctg tcaaagagaa agacatgacc aaggaattct ttgaaaacaa atcggaaaca 1860 tggccaatcg gagagtcacc caggggagtg gaggaaggct ctattgggaa agtgtgcagg 1920 accttactgg caaaatctgt attcaacagt ctatatgcgt ctccacaact tgaggggttt 1980 tcggctgaat Cgagaaaatt gcttctcatt gttcaggcac ttagggacaa cctggaacct 2040 ggaaccttcg atcttggggg gctatatgaa gcaatcgagg agtgcctgat taatgatccc 2100 tgggttttgc ttaatgcatc ttggttcaac tccttcctca cacatgcact gaagtagttg 2160 tggcaatgct ac 2172

&lt;210&gt; 28 &lt;211&gt; 1735 &lt;212&gt; DNA &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 28 aaaagcaaca aaagatgaag gcaatactag tagttctgct atatacattt gcaaccgcaa 60 at£cagacac attatgtata ggttatcatg cgaacaattc aacagacact gtagacacag 120 tactagaaaa gaatgtaaca gtaacacact ctgttaacct tctagaagac aagcataacg 180 g £ aaactatg caaactaaga ggggtagccc cattgcattt gggtaaatgt aacattgctg 240 gctggatcct gggaaatcca gagtgtgaat cactctccac agcaagctca tggtcctaca 300 ttgtggaaac atctagttca gacaatggaa cgtgttaccc aggagatttc atcgattatg 360 aggagctaag agagcaattg agctcagtgt catcatttga aaggtttgag atattcccca 420 agacaagttc atggcccaat catgactcga acaaaggtgt aacggcagca tgtcctcatg 480 ctggagcaaa aagcttctac aaaaatttaa tatggctagt taaaaaagga aattcatacc 540 caaagctcag caaatcctac attaatgata Aagggaaaga agtcctcgtg ctatggggca 600 ttcaccatcc atctactagt gctgaccaac aaagtctcta tcagaat£〇a gatgcatatg 660 tttttgtggg gtcatcaaga tacagcaaga agttcaagcc ggaaatagca ataagaccca 720 aagtgaggga tcaagaaggg agaatgaact attactggac actagtag ag ccgggagaca 780 aaataacatt cgaagcaact ggaaatctag tggtaccgag atatgcattc gcaatggaaa 840 gaaatgctgg atctggtatt atcatttcag atacaccagt ccacgattgc aatacaactt 900 gtcagacacc caagggtgct ataaacacca gcctcccatt tcagaatata catccgatca 960 caattggaaa atgtccaaaa tatgtaaaaa gcacaaaatt gagactggcc acaggattga 1020 ggaatgtccc gtctattcaa tctagaggcc tatttggggc cattgccggt ttcattgaag 1080 gggggtggac agggatggta gatggatggt acggttatca ccatcaaaat gagcaggggt 1140 -25 · 159890 · sEQUENCE LISTING .doc 201249996 caggatatgc agccgacctg aagagcacac agaatgccat tgacgagatt actaacaaag 1200 taaattctgt tattgaaaag atgaatacac a ^ ttcacagc agtaggtaaa gagttcaacc 1260 acctggaaaa aagaatagag aatttaaata aaaaagtt ^ a tgatggtttc ctggacattt 1320 ggacttacaa tgccgaactg ttggttctat tggaaaatga aagaactttg gactaccacg 1380 attcaaatgt gaagaactta tatgaaaagg taagaagcca gctaaaaaac aatgccaagg 1440 aaattggaaa cggctgcttt gaattttacc acaaatgcga taacacgtgc atggaaagtg 1500 tcaaaaatgg gacttatgac tacccaaaat actcagagga agcaaaatta aacagagaag 1560 aaatagatgg ggtaaagctg ga atcaacaa ggatttacca gattttggcg atctattcaa 1620 ctgtcgccag ttcattggta ctggtagtct ccctgggggc aatcagtttc tggatgtgct 1680 ctaatgggtc tctacagtgt agaatatgta tttaacttta ggatttcaga agcat 1735 &lt; 210 &gt; 29 &lt; 211 &gt; 1546 &lt; 212 &gt; Wk &lt; 213 &gt; H1N1 swine influenza virus &lt; 400 &gt; 29 atcaatcact gagtgacatc gaagccatgg cgtctcaagg caccaaacga tcatatgaac 60 aaatggagac tggtgg £ gag cgccaggatg ccacagaaat cagagcatct gtcggaagaa 120 tgattggtgg aatcgggaga ttctacatcc aaatgtgcac tgaactcaaa ctcagtgatt 180 atgatggacg actaatccag aatagcataa caatagagag gatggtgctt tctgcttttg 240 atgagagaag aaataaatac ctagaagagc atcccagtgc tgggaaggac cctaagaaaa 300 caggaggacc catatataga agaatagacg gaaagtggat gagagaactc atcctttatg 360 acaaagaaga aataaggaga gtttggcgcc aagcaaacaa tggcgaagat gcaacagcag 420 Gtcttactca tatcatgatt tggcattcca acctgaatga tgccacatat cagagaacaa 480 gagcgcttgt tcgcaccgga atg£atccca gaatgtgctc tctaatgcaa ggttcaacac 540 ttcccagaag gtctggtgcc gcaggtgctg cggtgaaagg agttggaaca atagcaatgg 600 agttaatcag a atgatcaaa cgtggaatca atgaccgaaa tttctggagg ggtgaaaatg 660 gacgaaggac aagggttgct tatgaaagaa tgtgcaatat cctcaaagga aaatttcaaa 720 cagctgccca gagggcaatg atggatcaag taagagaaag tcgaaaccca ggaaacgctg 780 agattgaaga cctcattttc ctggcacggt cagcactcat tctgagggga tcagttgcac 840 ataaatcctg cctgcctgct tgtgtgtatg ggcttgcagt agcaagtggg catgactttg 900 aaagggaagg gtactcactg gtcgggatag acccattcaa attactccaa aacagccaag 960 tggtcagcct gatgagacca aatgaaaacc cagctcacaa gagtcaattg gtgtggatgg 1020 catgccactc tgctgcattt gaagatttaa gagtatcaag tttcataaga ggaaagaaag 1080 tgattccaag aggaaagctt tccacaagag gggtccagat tgcttcaaat gagaatgtgg 1140 aaaccatgga ctccaatacc ctggaactaa gaagcagata ctgggccata aggaccagga 1200 gtggaggaaa taccaatcaa caaaaggcat ccgcaggcca gatcagtgtg cagcctacat 1260 tctcagtgca gcgaaatctc ccttttgaaa gagcaaccgt tatggcagca ttcagcggga 1320 acaatgaagg acggacatcc gacatgcgaa cagaagttat aagaatgatg gaaagtgcaa 1380 agccagaaga tttgtccttc caggggcggg gagtcttcga gctctcggac gaaaaggcaa 1440 • 26 · 159890·sequence .doc 201249996 cgaacccgat cgtgccttcc tttgacatga gtaatgaagg gtcttatuc ttcggagaca 1500 atgcagagga gtatgacagt tgaggaaaaa tacccttgtt tctact 1546 &lt;210&gt; 30 &lt;211&gt; 1410 &lt;212&gt; DNA &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 30 atgaatccaa accaaaagat aataaccatt ggttcggtct gtatgacgat tggaatggct 60 aacttaatat tacaaattgg aaacataatc tcaatatgga ttagccactc aattcaactt 120 gggaatcaaa atcagattga aacatgcaat caaagcgtca ttacttatga aaacaacact 180 tgggtaaatc agacatatgt taacatcagc aacaccaact ttgctgctgg acagtcagtg 240 gtttccgtga aattagcggg caattcctct ctctgccctg ttagtggatg ggctatatac 300 agtaaagaca acagtataag aatcggttcc aagggggatg tgtttgtcat aagggaacca 360 ttcatatcat gctccccctt ggaatgcaga accttcttct tgactcaagg ggccttgcta 420 aatgacaaac attccaatgg aaccattaaa gacaggagcc catatcgaac Cctaatgagc 480 tgtcctattg gtgaagttcc ctctccatac aactcaagat ttgagtcagt cgcttggtca 540 gcaagtgctt gtcatgatgg catcaattgg ctaacaattg gaatttctgg cccagacaat 600 ggggcagtgg ctgtgttaaa gtacaacggc ataataacag acactatcaa gagttggaga 660 aacaatatat tgagaacaca agagtctgaa tgtgcatgtg taaatggttc ttgctttact 720 gtaatgaccg atggaccaag tgatggacag gcctcataca agatcttcag aatagaaaag 780 .ggaaagatag tcaaatcagt cgaaatgaat gcccctaatt attactatga ggaatgctcc 840 tgttatcctg attctagtga aatcacatgt gtgtgcaggg ataactggca tggctcgaat 900 cgaccgtggg tgtctttcaa ccagaatctg gaatatcaga taggatacat atgcagtggg 960 attttcggag acaatccacg ccctaatgat aagacaggca gttgtggtcc agtatcgtct 1020 aatggagcaa atggagtaaa aggattttca ttcaaatacg gcaatggtgt ttggataggg 1080 agaactaaaa gcattagttc aagaaacggt tttgagatga tttgggatcc gaacggatgg 1140 actgggacag acaataactt ctcaataaag caagatatcg taggaataaa tgagtggtca 1200 ggatatagcg ggagttttgt tcagcatcca gaactaacag ggctggattg tataagacct 1260 tgcttctggg ttgaactaat cagagggcga cccaaagaga acacaatctg gactagcggg 1320 agcagcatat ccttttgtgg tgtaaacagt gacactgtgg gttggtcttg gccagacggt 1380 gctgagttgc catttaccat tgacaagtaa 1410 &lt; 210 &gt; 31 &lt; 211 &gt; 987 &lt;212&gt; DNA &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 31 taaagatgag tcttctaacc gaggtcgaaa cgtacgttct ttctatcatc ccgtcaggcc 60 ccctcaaagc cgagatcgcg cagagactgg aaagtgtctt tgcaggaaag aacacagatc 120 ttgaggctct catggaatgg ctaaagacaa gaccaatctt gtcacctctg actaagggaa 180 ttttaggatt tgtgttcacg ctcaccgtgc ccagtgagcg aggactgcag cgtagacgct 240 ttgtccaaaa tgccctaaat gggaatgggg acccgaacaa catggataga gcagttaaac 300 -27- 159890- Sequence Listing .doc 201249996 tatacaagaa gctcaaaaga gaaataacgt tccatggggc caaggaggtg tcactaagct 360 attcaactgg tgcacttgcc agttgcatgg gcctcatata caacaggatg ggaacagtga 420 ccacagaagc tgcttttggt ctagtgtgtg ccacttgtga acagattgct gattcacagc 480 atcsgtctca cagacagatg gctactacca ccaatccact aatcaggcat gaaaacagaa 540 tggtgctggc tagcactacg gcaaaggcta tggaacagat ggctggatcg agtgaacagg 600 cagcggaggc catggaggtt gctaatcaga ctaggcagat ggtacatgca atgagaacta 660 ttgggactca tcctagctcc agtgctggtc tgaaagatga ccttcttgaa aatttgcagg 720 cctaccagaa gcgaatggga gtgcagatgc agcgattcaa gtgatcctct Cgtcattgca 780 gcaaatatca ttgggatctt gcacctgata ttgtggatta ctgatcg Tct ttttttcaaa 840 tgtatttatc gtcgctttaa atacggtttg aaaagagggc cttctacgga aggagtgcct 900 gagtccatga gggaagaata tcaacaggaa cagcagagtg ctgtggatgt tgacgatggt 960 cattttgtca acatagagct agagtaa 987

&Lt; 210 &gt; 32 &lt; 211 &gt; 851 &lt; 212 &gt; DNA &lt; 213> H1N1 swine influenza virus &lt; 400 &gt; 32 aacataatgg actccaacac catgtcaagc tttcaggtag actgtttcct ttggcatatc 60 cgcaagcgat ttgcagacaa tggattgggt gatgccccat tccttgatcg gctccgccga 120 gatcaaaagt ccttaaaagg aagaggcaac acccttggcc tcgatatcga aacagccact 180 cttgttggga aacaaatcgt ggaatggatc ttgaaagagg aatccagcga gacacttaga 240 atgacaattg catctgtacc tacttcgcgc tacctttctg acatgaccct cgaggaaatg 300 tcacgagact ggttcatgct catgcctagg caaaa £ ataa taggccctct ttgcgtgcga 360 ttggaccagg cgatcatgga aaagaacata £ tactgaaag cgaacttcag tgtaatcttt 420 aaccgattag agaccttgat actactaagg gctttcactg aggagggagc aatagttgga 480 gaaatttcac cattaccttc tcttccagga catacttatg aggatgtcaa aaatgcaatt 540 ggggtcctca tcggaggact tgaatggaat ggtaacacgg Ttcgagtctc tgaaaatata 600 cagagattcg cttggagaaa ctgtgatgag aatgg£agac cttcactacc tccagagcag 660 aaatgaaaag tggcgagagc aattgggaca gaaatttgag gaaataaggt ggttaattga 720 agaaatgcgg cacagattga aagcgacaga gaatagtttc gaacaaataa ca Tttatgca 780 agccttacaa ctactgcttg aagtagaaca agagataaga gctttctcgt ttcagcttat 840 ttaactgata a 851

&lt;210&gt; 33 &lt;211&gt; 566 &lt;212&gt; PRT &lt;213&gt;Influenza A virus &lt;400&gt; 33

Met Lys Ala lie Leu Val Val Leu Leu Tyr Thr Phe Ala Thr Ala Asn 15 10 15

Ala Asp Thr Leu Cys lie Gly Tyr His Ala Asn Asn Ser Thr Asp Thr • 28· 159890 · Sequence Listing.doc 201249996 20 25 30

Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn 35 40 45

Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val 50 55 60

Ala Pro Leu His Leu Gly Lys Cys Asn lie Ala Gly Trp lie Leu Gly 65 70 75 80

Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr lie 85 90 95

Val Glu Thr Pro Ser Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe 100 105 110 lie Asp Tyr Glu Glu Leu Arg Glu Gin Leu Ser Ser Val Ser Ser Phe 115 120 125

Glu Arg Phe Glu lie Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp 130 135 140

Ser Asn Lys Gly Val Thr Ala Ala Cys Fro His Ala Gly Ala Lys Ser 145 150 155 160

Phe Tyr Lys Asn Leu lie Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro 165 170 175

Lys Leu Ser Lys Ser Tyr lie Asn Asp Lys Gly Lys Glu Val Leu Val 180 185 190

Leu Trp Gly lie His His Pro Ser Thr Ser Ala Asp Gin Gin Ser Leu 195 200 205

Tyr Gin Asn Ala Asp Ala Tyr Val Phe Val Gly Ser Ser Arg Tyr Ser 210 215 220

Lys Lys Phe Lys Pro Glu lie Ala lie Arg Pro Lys Val Arg Asp Gin 225 230 235 240

Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys 245 250 255

He Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe 260 265 270

Ala Met Glu Arg Asn Ala Gly Ser Gly lie lie lie Ser Asp Thr Pro 275 280 285

Val His Asp Cys Asn Thr Thr Cys Gin Thr Pro Lys Gly Ala lie Asn 290 295 300

Thr Ser Leu Pro Phe Gin Asn lie His Pro He Thr He Gly Lys Cys 305 310 315 320 -29- 159890 - Sequence Listing.doc 201249996

Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg 325 330 335

Asn He Pro Ser lie Gin Ser Arg Gly Leu Phe Gly Ala lie Ala Gly 340 345 350

Phe lie Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr 355 360 365

His His Gin Asn Glu Gin Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser 370 375 380

Thr Gin Asn Ala He Asp Glu lie Thr Asn Lys Val Asn Ser Val lie 385 390 395 400

Glu Lys Met Asn Thr Gin Phe Thr Ala Val Gly Lys Glu Phe Asn His 405 410 415

Leu Glu Lys Arg lie Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe 420 425 430

Leu Asp lie Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn 435 440 445

Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu 450 455 460

Lys Val Arg Ser Gin Leu Lys Asn Asn Ala Lys Glu lie Gly Asn Gly 465 470 475 480

Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Met Glu Ser Val 485 490 495

Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu 500 505 510

Asn Arg Glu Glu lie Asp Gly Val Lys Leu Glu Ser Thr Arg lie Tyr 515 520 525

Gin lie Leu Ala He Tyr Ser Thr Val Ala Ser Ser Leu Val Leu Val 530 535 540

Val Ser Leu Gly Ala lie Ser Phe Trp Met Cys Ser Asn Gly Ser Leu 545 550 555 560

Gin Cys Arg lie Cys lie 565

&lt;210&gt; 34 &lt;211&gt; 566 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 34

Met Lys Ala lie Leu Val Val Leu Leu Tyr Thr Phe Ala Thr Ala Asn 1 5 10 .15 •30· 159890-Sequence List.doc 201249996

Ala Asp Thr Leu Cys lie Gly Tyr His Ala Asn Asn Ser Thr Asp Thr 20 25 30

Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn 35 40 45

Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val 50 55 60

Ala Pro Leu His Leu Gly Lys Cys Asn lie Ala Gly Trp lie Leu Gly 65 70 75 80

Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr lie 85 90 95

Val Glu ΤΤ, γ Ser Ser Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe lie Asp Tyr Glu Glu Leu Arg Glu Gin Leu Ser Ser Val Ser Ser Phe 115 120 125

Glu Arg Phe Glu He Phe Pro Lys Tl, r Ser Ser Trp Pro Asp His Asp

Ser Asn Lys Gly Val ^ Ala Ala Cys Pro His Ala Gly A, a Lys Ser

Phe Tyr Lys Asn Leu lie Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro 165 170 175

Lys Leu Ser Lys Ser Tyr lie Asn Asp Lys Gly Lys Glu Val Leu Val 180 185 190

Leu Trp Gly lie His His Pro Ser Thr Ser Ala Asp Gin Gin Ser Leu 195 200 205

Tyr Gin Asn Ala Asp Ala Tyr Val Phe Val Gly Thr Ser Arg Tyr Ser 210 215 220

Lys Lys Phe Lys Pro Glu lie Ala lie Arg Pro Lys Val Arg Glu Arg 225 230 235 240

Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys 245 250 255 lie Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe 260 265 270

Ala Met Glu Arg Asn Ala Gly Ser Gly lie He lie Ser Asp Thr Pro 275 280 285

Val His Asp Cys Asn Thr Thr Cys Gin Thr Pro Lys Gly Ala lie Asn 290 295 300

Thr Ser Leu Pro Phe Gin Asn lie His Pro lie Thr lie Gly Lys Cys 305 310 315 320 • 31- 159890 - Sequence Listing.doc 201249996

Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg 325 330 335

Asn Val Pro Ser lie Gin Ser Arg Gly Leu Phe Gly Ala lie Ala Gly 340 345 350

Phe lie Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr 355 360 365

His His Gin Asn Glu Gin Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser 370 375 380

Thr Gin Asn Ala lie Asp Glu lie Thr Asn Lys Val Asn Ser Val lie 385 390 395 400 G】u Lys Met Asn Thr Gin Phe Thr Ala Va】G】y Lys Glu Phe Asn His 405 410 415

Leu Glu Lys Arg lie Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe 420 425 430

Leu Asp lie Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn 435 440 445

Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu 450 455 460

Lys Val Arg Ser Gin Leu Lys Asn Asn Ala Lys Glu lie Gly Asn Gly 465 470 475 480

Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Met Glu Ser Val 485 490 495

Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu 500 505 510

Asn Arg Glu Glu lie Asp Gly Val Lys Leu Glu Ser Thr Arg lie Tyr 515 520 525

Gin lie Leu Ala lie Tyr Ser Thr Val Ala Ser Ser Leu Val Leu Val 530 535 540

Val Ser Leu Gly Ala lie Ser Phe Trp Met Cys Ser Asn Gly Ser Leu 545 550 555 560

Gin Cys Arg lie Cys lie 565

&lt;210&gt; 35 &lt;211&gt; 566 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 35

Met Lys Ala lie Leu Val Val Leu Leu Tyr Thr Phe Ala Thr Ala Asn 15 10 15 •32- 159890-Sequence List.doc 201249996

Ala Asp Thr Leu Cys lie Gly Tyr His Ala Asn Asn Ser TTir Asp Thr 20 25 30

Val Asp Thr Val Leu Glu Lys Asn Va] Thr Val Thr His Ser Val Asn 35 40 45

Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val 50 55 60

Ala Pro Leu His Leu Gly Lys Cys Asn lie Ala Gly Trp lie Leu Gly 65 70 75 80

Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr lie 85 90 95

Val Glu Thr Ser Ser Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe 100 105 110 lie Asp Tyr Glu Glu Leu Arg Glu Gin Leu Ser Ser Val Ser Ser Phe 115 120 125

Glu Arg Phe Glu lie Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp 130 135 140

Ser Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser 145 150 155 160

Phe Tyr Lys Asn Leu lie Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro 165 170 175

Lys Leu Ser Lys Ser Tyr He Asn Asp Lys Gly Lys Glu Val Leu Val 180 185 190

Leu Trp Gly lie His His Pro Ser Thr Ser Ala Asp Gin Gin Ser Leu 195 200 205

Tyr Gin Asn Ala Asp Ala Tyr Val Phe Val Gly Ser Ser Arg Tyr Ser 210 215 220

Lys Lys Phe Lys Pro Glu lie Ala lie Arg Pro Lys Val Arg Asp Gin 225 230 235 240

Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys 245 250 255 lie Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe 260 265 270

Ala Met Glu Arg Asn Ala Gly Ser Gly He lie lie Ser Asp Thr Pro 275 280 285

Val His Asp Cys Asn Thr Thr Cys Gin Thr Pro Lys Gly Ala lie Asn 290 295 300

Thr Ser Leu Pro Phe Gin Asn lie His Pro He Thr lie Gly Lys Cys 305 310 315 320 •33· 159890-Sequence List.d〇c 201249996

Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg 325 330 335

Asn Val Pro Ser lie Gin Ser Arg Gly Leu Phe Gly Ala lie Ala Gly 340 345 350

Phe lie Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr 355 360 365

His His Gin Asn Glu Gin Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser 370 375 380

Thr Gin Asn Ala lie Asp Glu lie Thr Asn Lys Val Asn Ser Val lie 385 390 395 400

Glu Lys Met Asn Thr Gin Phe Thr Ala Val Gly Lys Glu Phe Asn His 405 410 415

Leu Glu Lys Arg I】e Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe 420 425 430

Leu Asp lie Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn 435 440 445

Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu 450 455 460

Lys Val Arg Ser Gin Leu Lys Asn Asn Ala Lys Glu lie Gly Asn Gly 465 470 475 480

Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Met Glu Ser Val 485 490 495

Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu 500 505 510

Asn Arg Glu Glu lie Asp Gly Val Lys Leu Glu Ser Thr Arg lie Tyr 515 520 525

Gin lie Leu Ala lie Tyr Ser Thr Val Ala Ser Ser Leu Val Leu Val 530 535 540

Val Ser Leu Gly Ala lie Ser Phe Trp Met Cys Ser Asn Gly Ser Leu 545 550 555 560

Gin Cys Arg He Cys lie 565

&lt;210&gt; 36 &lt;211&gt; 566 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 36

Met Lys Ala He Leu Val Val Leu Leu Tyr Thr Phe Ala Thr Ala Asn 15 10 15 -34- 159890 · Sequence Listing.doc 201249996

Ala Asp Thr Leu Cys lie Gly Tyr His Ala Asn Asn Ser Thr Asp Thr 20 25 30

Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn 35 40 45

Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val 50 55 60

Ala Pro Leu His Leu Gly Lys Cys Asn lie Ala Gly Trp lie Leu Gly 65 70 75 g〇

Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Scr Ser Trp Ser Tyr lie 85 90 95

Val Glu TTir Ser Ser Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe 100 105 110 lie Asp Tyr Glu Glu Leu Arg Glu Gin Leu Ser Ser Val Ser Ser Phe 115 120 125

Glu Arg Phe Glu lie Phe Pro Lys Thr Ser SeT Trp Pro Asn His Asp 130 135 140

Ser Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser 145 150 155 160

Phe Tyr Lys Asn Leu lie Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro 165 170 175

Lys Leu Ser Lys Ser Tyr lie Asn Asp Lys Gly Lys Glu Val Leu Val 180 185 190

Leu Trp Gly lie His His Pro Ser Thr Ser Ala Asp Gin Gin Ser Leu 195 200 205

Tyr Gin Asn Ala Asp Ala Tyr Val Phe Val Gly Ser Ser Arg Tyr Ser 210 215 220

Lys Lys Phe Lys Pro Glu lie Ala lie Arg Pro Lys Val Arg Asp Gin 225 230 235 240

Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys 245 250 255 lie Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe 260 265 270

Ala Met Glu Arg Asn Ala Gly Ser Gly lie lie lie Ser Asp Thr Pro 275 280 285

Val His Asp Cys Asn Thr Thr Cys Gin Thr Pro Lys Gly Ala lie Asn 290 295 300

Thr Ser Leu Pro Phe Gin Asn lie His Pro lie Thr lie Gly Lys Cys -35- 159890 - Sequence Listing.d〇c 201249996 305 310 315 320

Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg 325 330 335

Asn Val Pro Ser lie Gin Ser Arg Gly Leu Phe Gly Ala lie Ala Gly 340 345 350

Phe lie Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr 355 360 365

His His Gin Asn Glu Gin Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser 370 375 380

Thr Gin Asn Ala lie Asp Glu lie Thr Asn Lys Val Asn Ser Val lie 385 390 395 400

Glu Lys Met Asn Thr Gin Phe Thr Ala Val Gly Lys Glu Phe Asn His 405 410 415

Leu Glu Lys Arg He Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe 420 425 430

Leu Asp lie Ττρ Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn 435 440 445

Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu 450 455 460

Lys Val Arg Ser Gin Leu Lys Asn Asn Ala Lys Glu lie Gly Asn Gly 465 470 475 480

Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Met Glu Ser Val 485 490 495

Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu 500 505 510

Asn Arg G】u Glu lie Asp Gly Val Lys Leu Glu Ser Thr Arg I】e Tyr 515 520 525

Gin lie Leu Ala lie Tyr Ser Thr Val Ala Ser Ser Leu Val Leu Val 530 535 540

Val Ser Leu Gly Ala lie Ser Phe Trp Met Cys Ser Asn Gly Ser Leu 545 550 555 560

Gin Cys Arg lie Cys lie 565

&lt;210&gt; 37 &lt;211&gt; 566 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 37

Met Lys Ala lie Leu Val Val Leu Leu Tyr Thr Phe Ala Thr Ala Asn -36- 159890 - Sequence Listing.doc 201249996 15 10 15

Ala Asp Thr Leu Cys lie Gly Tyr His Ala Asn Asn Ser Thr Asp Thr 20 25 30

Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn 35 40 45

Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val 50 55 60

Ala Pro Leu His Leu Gly Lys Cys Asn lie Ala Gly Trp lie Leu Gly 65 70 75 80

Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr lie 85 90 95

Val Glu Thr Ser Ser Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe 100 105 110 lie Asp Tyr Glu Glu Leu Arg Glu Gin Leu Ser Ser Val Ser Ser Phe 115 120 125

Glu Arg Phe Glu lie Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp 130 135 140

Ser Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser 145 150 155 160

Phe Tyr Lys Asn Leu lie Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro 165 170 175

Lys Leu Ser Lys Ser Tyr lie Asn Asp Lys Gly Lys Glu Val Leu Val 180 185 190

Leu Trp Gly lie His His Pro Ser Thr Ser Ala Asp Gin Gin Ser Leu 195 200 205

Tyr Gin Asn Ala Asp Ala Tyr Val Phe Val Gly Ser Ser Arg Tyr Ser 210 215 220

Lys Lys Phe Lys Pro Glu lie Ala lie Arg Pro Lys Val Arg Asp Gin 225 230 235 240

Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys 245 250 255 lie Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe 260 265 270

Ala Met Glu Arg Asn Ala Gly Ser Gly lie lie lie Ser Asp Thr Pro 275 280 285

Val His Asp Cys Asn Thr Thr Cys Gin Thr Pro Lys Gly Ala lie Asn 290 295 300 • 37· 159890 - Sequence Listing.doc 201249996

Thr Ser Leu Pro Phe Gin Asn lie His Pro lie Thr lie Gly Lys Cys 305 310 315 320

Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg 325 330 335

Asn Val Pro Ser lie Gin Ser Arg Gly Leu Phe Gly Ala lie Ala Gly 340 345 350

Phe lie Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr 355 360 365

His His Gin Asn Glu Gin Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser 370 375 380

Thr Gin Asn Ala lie Asp Glu lie Thr Asn Lys Val Asn Ser Val lie 385 390 395 400

Glu Lys Met Asn Thr Gin Phe Thr Ala Val Gly Lys Glu Phe Asn His 405 410 415

Leu Glu Lys Arg lie Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe 420 425 430

Leu Asp lie Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn 435 440 445

Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu 450 455 460

Lys Val Arg Ser Gin Leu Lys Asn Asn Ala Lys Glu lie Gly Asn Gly 465 470 475 480

Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Met Glu Ser Val 485 490 495

Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu 500 505 510

Asn Arg Glu Glu He Asp Gly Val Lys Leu Glu Ser Thr Arg lie Tyr 515 520 525

Gin He Leu Ala lie Tyr Ser Thr Val Ala Ser Ser Leu Val Leu Val 530 535 540

Val Ser Leu Gly Ala lie Ser Phe Trp Met Cys Ser Asn Gly Ser Leu 545 550 555 560

Gin Cys Arg lie Cys lie 565

&lt;210&gt; 38 &lt;211&gt; 469 &lt;212&gt; PRT &lt;213&gt;Influenza A virus &lt;400&gt; 38 • 38 · 159890 - Sequence Listing.doc 201249996

Met Asn Pro Asn Gin Lys lie lie Thr lie Gly Ser Val Cys Met Thr 15 10 15 lie Gly Met Ala Asn Leu lie Leu Gin lie Gly Asn lie lie Ser lie 20 25 30

Trp lie Ser His Ser lie Gin Leu Gly Asn Gin Asn Gin lie Glu Thr 35 40 45

Cys Asn Gin Ser Val lie Thr Tyr Glu Asn Asn Thr Trp Val Asn Gin 50 55 60

Thr Tyr Val Asn lie Ser Asn Thr Asn Phe Ala Ala Gly Gin Ser Val 65 70 75 80

Val Ser Val Lys Leu Ala Gly Asn Ser Ser Leu Cys Pro Val Ser Gly 85 90 95

Trp Ala lie Tyr Ser Lys Asp Asn Ser Val Arg lie Gly Ser Lys Gly 100 105 110

Asp Val Phe Val lie Arg Glu Pro Phe lie Ser Cys Ser Pro Leu Glu 115 120 125

Cys Arg Thr Phe Phe Leu Thr Gin Gly Ala Leu Leu Asn Asp Lys His 130 135 140

Ser Asn Gly Thr lie Lys Asp Arg Ser Pro Tyr Arg Thr Leu Met Ser 145 150 155 160

Cys Pro lie Gly Glu Val Pro Ser Pro Tyr Asn Ser Ar£ Phe Glu Ser 165 170 175

Val Ala Trp Ser Ala Ser Ala Cys His Asp Gly lie Asn Trp Leu Thr 180 185 190 lie Gly lie Ser Gly Pro Asp Asn Gly Ala Val Ala Val Leu Lys Tyr 195 200 205

Asn Gly lie lie Thr Asp Thr He Lys Ser Trp Arg Asn Asn He Leu 210 215 220

Arg Thr Gin Glu Ser Glu Cys Ala Cys Val Asn Gly Ser Cys Phe Thr 225 230 235 240

Val Met Thr Asp Gly Pro Ser Asn Gly Gin Ala Ser Tyr Lys lie Phe 245 250 255

Arg lie Glu Lys Gly Lys lie Val Lys Ser Val Glu Met Asn Ala Pro 260 265 270

Asn Tyr His Tyr Glu Glu Cys Ser Cys Tyr Pro Asp Ser Ser Glu lie 275 280 285

Thr Cys Val Cys Arg Asp Asn Trp His Gly Ser Asn Arg Pro Trp Val 290 295 300 • 39· 159890 - Sequence Listing doc 201249996

Ser Phe Asn Gin Asn Leu Glu Tyr Gin He Gly Tyr lie Cys Ser Gly 305 310 315 320 lie Phe Gly Asp Asn Pro Arg Pro Asn Asp Lys Thr Gly Ser Cys Gly 325 330 335

Pro Val Ser Ser Asn Gly Ala Asn Gly Val Lys Gly Phe Ser Phe Lys 340 345 350

Tyr Gly Asn Gly Val Trp lie Gly Arg Thr Lys Ser lie Ser Ser Arg 355 360 365

Asn Gly Phe Glu Met lie Trp Asp Pro Asn Gly Trp Thr Gly Thr Asp 370 375 380

Asn Asn Phe Ser lie Lys Gin Asp lie Val Gly lie Asn Glu Trp Ser 385 390 395 400

Gly Tyr Ser Gly Ser Phe Val Gin His Pro Glu Leu Thr Gly Leu Asp 405 410 415

Cys lie Arg Pro Cys Phe Trp Val Glu Leu lie Arg Gly Arg Pro Lys 420 425 430

Glu Asn Thr lie Trp Thr Ser Gly Ser Ser lie Ser Phe Cys Gly Val 435 440 445

Asn Ser Asp Thr Val Gly Trp Ser Trp Pro Asp Gly Ala Glu Leu Pro 450 455 460

Phe Thr lie Asp Lys 465

&lt;210&gt; 39 &lt;211&gt; 469 &lt;212&gt; PRT &lt;213>H1N1 swine influenza virus &lt;400&gt; 39

Met Asn Pro Asn Gin Lys He lie Thr lie Gly Ser Val Cys Met Thr 15 10 15 lie Gly Met Ala Asn Leu He Leu Gin lie Gly Asn lie lie Ser He 20 25 30

Trp lie Ser His Ser lie Gin Leu Gly Asn Gin Asn Gin lie Glu Thr 35 40 45

Cys Asn Gin Ser Val lie Thr Tyr Glu Asn Asn Thr Trp Val Asn Gin 50 55 60

Thr Tyr Val Asn lie Ser Asn Thr Asn Phe Ala Ala Gly Gin Ser Val 65 70 75 80

Val Ser Val Lys Leu Ala Gly Asn Ser Ser Leu Cys Pro Val Ser Gly 85 90 95 •40- 159890 · Sequence Listing.doc 201249996

Trp Ala lie Tyr Ser Lys Asp Asn Ser lie Arg lie Gly Ser Lys Gly 100 105 110

Asp Val Phe Val lie Arg Glu Pro Phe He Ser Cys Ser Pro Leu Glu 115 120 125

Cys Arg Thr Phe Phe Leu Thr Gin Gly Ala Leu Leu Asn Asp Lys His 130 135 140

Ser Asn Gly Thr lie Lys Asp Arg Ser Pro Tyr Arg Thr Leu Met Ser 145 150 155 160

Cys Pro lie Gly Glu Val Pro Ser Pro Tyr Asn Ser Arg Phe Glu Ser 165 170 175

Val Ala Trp Ser Ala Ser Ala Cys His Asp Gly lie Asn Trp Leu Thr 180 185 190 lie Gly lie Ser Gly Pro Asp Asn Gly Ala Val Ala Val Leu Lys Tyr 195 200 205

Asn Gly He lie Thr Asp Thr lie Lys Ser Trp Arg Asn Asn lie Leu 210 215 220

Arg Thr Gin Glu Ser Glu Cys Ala Cys Val Asn Gly Ser Cys Phe Thr 225 230 235 240

Val Met Thr Asp Gly Pro Ser Asp Gly Gin Ala Ser Tyr Lys lie Phe 245 250 255

Arg He Glu Lys Gly Lys lie Val Lys Ser Val Glu Met Asn Ala Pro 260 265 270

Asn Tyr Tyr Tyr Glu Glu Cys Ser Cys Tyr Pro Asp Ser Ser Glu lie 275 280 285

Thr Cys Val Cys Arg Asp Asn Trp His Gly Ser Asn Arg Pro Trp Val 290 295 300

Ser Phe Asn Gin Asn Leu Glu Tyr Gin lie Gly Tyr lie Cys Ser Gly 305 310 315 320

He Phe Gly Asp Asn Pro Arg Pro Asn Asp Lys Thr Gly Ser Cys Gly 325 330 335

Pro Val Ser Ser Asn Gly Ala Asn Gly Val Lys Gly Phe Ser Phe Lys 340 345 350

Tyr Gly Asn Gly Val Trp lie Gly Arg Thr Lys Ser lie Ser Ser Arg 355 360 365

Asn Gly Phe Glu Met lie Trp Asp Pro Asn Gly Trp Thr Gly Thr Asp 370 375 380

Asn Asn Phe Ser lie Lys Gin Asp lie Val Gly lie Asn Glu Trp Ser 385 390 395 400 -41 - 159890 - Sequence Listing _doc 201249996

Gly Tyr Ser Gly Ser Phe Val Gin His Pro Glu Leu Thr Gly Leu Asp 405 410 415

Cys lie Arg Pro Cys Phe Trp Val Glu Leu lie Arg Gly Arg Pro Lys 420 425 430

Glu Asn Thr lie Trp Thr Ser Gly Ser Ser lie Ser Phe Cys Gly Val 435 440 445

Asn Ser Asp Thr Val Gly Trp Ser Trp Pro Asp Gly Ala Glu Leu Pro 450 455 460

Phe Thr lie Asp Lys 465

&lt;210&gt; 40 &lt;211&gt; 469 &lt;212&gt; PRT &lt;213&gt; H1N] Swine Influenza Virus &lt;400&gt; 40

Met Asn Pro Asn Gin Lys lie lie Thr lie Gly Ser Val Cys Met Thr 15 10 15 lie Gly Met Ala Asn Leu lie Leu Gin lie Gly Asn lie lie Ser lie 20 25 30

Trp lie Ser His Ser lie Gin Leu Gly Asn Gin Asn Gin lie Glu Thr 35 40 45

Cys Asn Gin Ser Val lie Thr Tyr Glu Asn Asn Thr Trp Val Asn Gin 50 55 60

Thr Tyr Val Asn lie Ser Asn Thr Asn Phe Ala Ala Gly Gin Ser Val 65 70 75 80

Val Ser Val Lys Leu Ala Gly Asn Ser Ser Leu Cys Pro Val Ser Gly 85 90 95

Trp Ala lie Tyr Ser Lys Asp Asn Ser lie Arg lie Gly Ser Lys Gly 100 105 110

Asp Val Phe Val lie Arg Glu Pro Phe lie Ser Cys Ser Pro Leu Glu 115 120 125

Cys Arg Thr Phe Phe Leu Thr Gin Gly Ala Leu Leu Asn Asp Lys His 130 135 140

Ser Asn Gly Thr lie Lys Asp Arg Ser Pro Tyr Arg Thr Leu Met Ser 145 150 155 160

Cys Pro lie Gly Glu Val Pro Ser Pro Tyr Asn Ser Arg Phe Glu Ser 165 170 175

Val Ala Trp Ser Ala Ser Ala Cys His Asp Gly lie Asn Trp Leu Thr 180 185 190 • 42- 159890 · Sequence Listing.doc 201249996 lie Gly lie Ser Gly Pro Asp Asn Gly Ala Val Ala Val Leu Lys Tyr 195 200 205

Asn Gly lie lie Thr Asp Thr He Lys Ser Trp Arg Asn Asn lie Leu 210 215 220

Arg Thr Gin Glu Ser Glu Cys Ala Cys Val Asn Gly Ser Cys Phe Thr 225 230 235 240

Val Met Thr Asp Gly Pro Ser Asp Gly Gin Ala Ser Tyr Lys lie Phe 245 250 255

Arg lie Glu Lys Gly Lys lie Val Lys Ser Val Glu Met Asn Ala Pro 260 265 270

Asn Tyr Tyr Tyr Glu Glu Cys Ser Cys Tyr Pro Asp Ser Ser Glu He 275 280 285

Thr Cys Val Cys Arg Asp Asn Trp His Gly Ser Asn Arg Pro Trp Val 290 295 300

Ser Phe Asn Gin Asn Leu Glu Tyr Gin lie Gly Tyr lie Cys Ser Gly 305 310 315 320 lie Phe Gly Asp Asn Pro Arg Pro Asn Asp Lys Thr Gly Ser Cys Gly 325 330 335

Pro Val Ser Ser Asn Gly Ala Asn Gly Val Lys Gly Phe Ser Phe Lys 340 345 350

Tyr Gly Asn Gly Val Trp lie Gly Arg Thr Lys Ser lie Ser Ser Arg 355 360 365

Asn Gly Phe Glu Met Tie Trp Asp Pro Asn Gly Trp Thr Gly Thr Asp 370 375 380

Asn Asn Phe Ser lie Lys Gin Asp lie Val Gly He Asn Glu Trp Ser 385 390 395 400

Gly Tyr Ser Gly Ser Phe Val Gin His Pro Glu Leu Thr Gly Leu Asp 405 410 415

Cys lie Arg Pro Cys Phe Trp Val Glu Leu lie Arg Gly Arg Pro Lys 420 425 430

Glu Asn Thr lie Trp Thr Ser Gly Ser Ser lie Ser Phe Cys Gly Val 435 440 445

Asn Ser Asp Thr Val Gly Trp Ser Trp Pro Asp Gly Ala Glu Leu Pro 450 455 460

Phe Thr He Asp Lys 465 &lt;210&gt; 41 •43- 159890 · Sequence Listing.doc 201249996

&lt;211&gt; 469 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 41

Met Asn Pro Asn Gin Lys lie lie Thr lie Gly Ser Va] Cys Met Thr 15 10 15 lie Gly Met Ala Asn Leu Met Leu Gin lie Gly Asn lie lie Ser lie 20 25 30

Trp lie Ser His Ser lie Gin Leu Gly Asn Gin Asn Gin lie Glu Thr 35 40 45

Cys Asn Gin Ser Val lie Thr Tyr Glu Asn Asn Thr Trp Val Asn Gin 50 55 60

Thr Tyr Val Asn lie Ser Asn Thr Asn Phe Ala Ala Gly Gin Ser Val 65 70 75 80

Val Ser Val Lys Leu Ala Gly Asn Ser Ser Leu Cys Pro Val Ser Gly 85 90 95

Trp Ala lie Tyr Ser Lys Asp Asn Ser Val Arg lie Gly Ser Lys Gly 100 105 110

Asp Val Phe Val lie Arg Glu Pro Phe lie Ser Cys Ser Pro Leu Glu 115 120 125

Cys Arg Thr Phe Phe Leu Thr Gin Gly Ala Leu Leu Asn Asp Lys His 130 135 140

Ser Asn Gly Thr lie Lys Asp Arg Ser Pro Tyr Arg Thr Leu Met Ser 145 150 155 160

Cys Pro lie Gly Glu Val Pro Ser Pro Tyr Asn Ser Arg Phe Glu Ser 165 170 175

Val Ala Trp Ser Ala Ser Ala Cys His Asp Gly lie Asn Trp Leu Thr 180 185 190 lie Gly lie Ser Gly Pro Asp Asn Gly Ala Val Ala Val Leu Lys Tyr 195 200 205

Asn Gly lie lie Thr Asp Thr lie Lys Ser Trp Arg Asn Asn lie Leu 210 215 220

Arg Thr Gin Glu Ser Glu Cys Ala Cys Val Asn Gly Ser Cys Phe Thr 225 230 235 240

Val Met Thr Asp Gly Pro Ser Asn Glu Gin Ala Ser Tyr Lys lie Phe 245 250 255

Arg lie Glu Lys Gly Lys· lie Val Lys Ser Val Glu Met Asn Ala Pro 260 265 270

Asn Tyr His Tyr Glu Glu Cys Ser Cys Tyr Pro Asp Ser Ser Glu lie -44- 159890 · Sequence Listing.doc 201249996 275 280 285

Thr Cys Val Cys Arg Asp Asn Trp His Gly Ser Asn Arg Pro Trp Val 290 295 300

Ser Phe Asn Gin Asn Leu Glu Tyr Gin lie Gly Tyr lie Cys Ser Gly 305 310 315 320 lie Phe Gly Asp Asn Pro Arg Pro Asn Asp Lys Thr Gly Ser Cys Gly 325 330 335

Pro Val Ser Ser Asn Gly Ala Asn Gly Val Lys Gly Phe Ser Phe Lys 340 345 350

Tyr Gly Asn Gly Val Trp lie Gly Arg Thr Lys Ser lie Ser Ser Arg 355 360 365

Asn Gly Phe Glu Met lie Trp Asp Pro Asn Gly Trp Thr Gly Thr Asp 370 375 380

Asn Asn Phe Ser lie Lys Gin Asp lie Val Gly lie Asn Glu Trp Ser 385 390 395 400

Gly Tyr Ser Gly Ser Phe Val Gin His Pro Glu Leu Thr Gly Leu Asp 405 410 415

Cys lie Arg Pro Cys Phe Trp Val Glu Leu lie Arg Gly Arg Pro Lys 420 425 430

Glu Asn Thr lie Trp Thr Ser Gly Ser Ser lie Ser Phe Cys Gly Val 435 440 445

Asn Ser Asp Thr Val Gly Trp Ser Trp Pro Asp Gly Ala Glu Leu Pro 450 455 460

Phe Thr lie Asp Lys 465 &lt;210&gt; 42 &lt;211&gt; 469 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt;

Met Asn Pro Asn Gin Lys lie lie Thr lie Gly Ser Val Cys Met Thr 15 10 15 lie Gly Met Ala Asn Leu lie Leu Gin lie Gly Asn lie lie Ser lie 20 25 30

Trp lie Ser His Ser lie Gin Leu Gly Asn Gin Asn Gin lie Glu Thr 35 40 45

Cys Asn Gin Ser Val lie Thr Tyr Glu Asn Asn Thr Trp Val Asn Gin 50 55 60

Thr Tyr Val Asn lie Ser Asn Thr Asn Phe Ala Ala Gly Gin Ser Val • 45· 159890 · Sequence Listing.doc 201249996 65 70 75 80

Val Ser Val Lys Leu Ala Gly Asn Ser Ser Leu Cys Pro Val Ser Gly 85 90 95

Trp Ala lie Tyr Ser Lys Asp Asn Ser lie Arg lie Gly Ser Lys Gly 100 105 110

Asp Val Phe Val lie Arg Glu Pro Phe lie Ser Cys Ser Pro Leu Glu 115 120 125

Cys Arg Thr Phe Phe Leu Thr Gin Gly Ala Leu Leu Asn Asp Lys His 130 135 140

Ser Asn Gly Thr lie Lys Asp Arg Ser Pro Tyr Arg Thr Leu Met Ser 145 150 155 160

Cys Pro He Gly Glu Val Pro Ser Pro Tyr Asn Ser Arg Phe Glu Ser 165 170 175

Val Ala Trp Ser Ala Ser Ala Cys His Asp Gly lie Asn Trp Leu Thr 180 185 190 lie Gly lie Ser Gly Pro Asp Asn Giy Ala Val Ala Val Leu Lys Tyr 195 200 205

Asn Gly lie lie Thr Asp Thr lie Lys Ser Trp Arg Asn Asn lie Leu 210 215 220

Arg Thr Gin Glu Ser Glu Cys Ala Cys Val Asn Gly Ser Cys Phe Thr 225 230 235 240

Val Met Thr Asp Gly Pro Ser Asp Gly Gin Ala Ser Tyr Lys lie Phe 245 250 255

Arg lie Glu Lys Gly Lys lie Val Lys Ser Val Glu Met Asn Ala Pro 260 265 270

Asn Tyr Tyr Tyr Glu Glu Cys Ser Cys Tyr Pro Asp Ser Ser Glu lie 275 280 285

Thr Cys Val Cys Arg Asp Asn Trp His Gly Ser Asn Arg Pro Trp Val 290 295 300

Ser Phe Asn Gin Asn Leu Glu Tyr Gin lie Gly Tyr lie Cys Ser Gly 305 310 315 320 lie Phe Gly Asp Asn Pro Arg Pro Asn Asp Lys Thr Gly Ser Cys Gly 325 330 335

Pro Val Ser Ser Asn Gly Ala Asn Gly Val Lys Gly Phe Ser Phe Lys 340 345 350

Tyr Gly Asn Gly Val Trp lie Gly Arg Thr Lys Ser lie Ser Ser Arg 355 360 365 46- 159890 · Sequence Listing.doc 201249996

Asn Gly Phe Glu Met lie Trp Asp Pro Asn Gly Trp Thr Gly Thr Asp 370 375 380

Asn Asn Phe Ser lie Lys Gin Asp lie Val Gly lie Asn Glu Trp Ser 385 390 395 400

Gly Tyr Ser Gly Ser Phe Val Gin His Pro Glu Leu Thr Gly Leu Asp 405 410 415

Cys lie Arg Pro Cys Phe Ττρ Val Glu Leu lie Arg Gly Arg Pro Lys 420 425 430

Glu Asn Thr lie Trp Thr Ser Gly Ser Ser lie Ser Phe Cys Gly Val 435 440 445

Asn Ser Asp Thr Val Gly Trp Ser Trp Pro Asp Gly Ala Glu Leu Pro 450 455 460

Phe Thr He Asp Lys 465

&lt;210&gt; 43 &lt;211&gt; 252 &lt;212&gt; PRT &lt;213&gt; Influenza A virus &lt;400&gt;

Met Ser Leu Leu Thr Glu Val Glu Thr Tyr Val Leu Ser lie lie Pro 15 10 15

Ser Gly Pro Leu Lys Ala Glu lie Ala Gin Arg Leu Glu Ser Val Phe 20 25 30

Ala Gly Lys Asn Thr Asp Leu Glu Ala Leu Met Glu Trp Leu Lys Thr 35 40 45

Arg Pro lie Leu Ser Pro Leu Thr Lys Gly lie Leu Gly Phe Val Phe 50 55 60

Thr Leu Thr Val Pro Ser Glu Arg Gly Leu Gin Arg Arg Arg Phe Val 65 70 75 80

Gin Asn Ala Leu Asn Gly Asn Gly Asp Pro Asn Asn Met Asp Arg Ala 85 90 95

Val Lys Leu Tyr Lys Lys Leu Lys Arg Glu He Thr Phe His Gly Ala 100 105 110

Lys Glu Val Ser Leu Ser Tyr Ser Thr Gly Ala Leu Ala Ser Cys Met 115 120 125

Gly Leu lie Tyr Asn Arg Met Gly Thr Val Thr Thr Glu Ala Ala Phe 130 135 140

Gly Leu Val Cys Ala Thr Cys Glu Gin lie Ala Asp Ser Gin His Arg 145 150 155 160 -47- 159890 · Sequence Listing.doc 201249996

Ser His Arg Gin Met Ala Thr Thr Thr Asn Pro Leu lie Arg His Glu 165 170 175

Asn Arg Met Val Leu Ala Ser Thr Thr Ala Lys Ala Met Glu Gin Met 180 185 190

Ala Gly Ser Ser Glu Gin Ala Ala Glu Ala Met Glu Val Ala Asn Gin 195 200 205

Thr Arg Gin Met Val His Ala Met Arg Thr lie Gly Thr His Pro Ser 210 215 220

Ser Ser Ala Gly Leu Lys Asp Asp Leu Leu Glu Asn Leu Gin Ala Tyr 225 230 235 240

Gin Lys Arg Met Gly Val Gin Met Gin Arg Phe Lys 245 250

&lt;210&gt; 44 &lt;211&gt; 97 &lt;212&gt; PRT &lt;213&gt; Influenza A virus &lt;400&gt; 44

Met Ser Leu Leu Thr Glu Val Glu TTir Pro Thr Arg Ser Glu Trp Glu 15 10 15

Cys Arg Cys Ser Asp Ser Ser Asp Pro Leu Val lie Ala Ala Asn He 20 25 30 lie Gly lie Leu His Leu lie Leu Trp lie Thr Asp Arg Leu Phe Phe 35 40 45

Lys Cys lie Tyr Arg Arg Phe Lys Tyr Gly Leu Lys Arg Gly Pro Ser 50 55 60

Thr Glu Gly Val Pro Glu Ser Met Arg Glu Glu Tyr Gin Gin Glu Gin 65 70 75 80

Gin Ser Ala Val Asp Val Asp Asp Gly His Phe Val Asn lie Glu Leu 85 90 95

Glu

&lt;210&gt; 45 &lt;211&gt; 252 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 45

Met Ser Leu Leu Thr Glu Val Glu Thr Tyr Val Leu Ser lie lie Pro 15 10 15

Ser Gly Pro Leu Lys Ala Glu lie Ala Gin Arg Leu Glu Ser Val Phe 20 25 30

Ala Gly Lys Asn Thr Asp Leu Glu Ala Leu Met Glu Trp Leu Lys Thr • 48- 159890 - Sequence Listing.doc 201249996 35 40 45

Arg Pro lie Leu Ser Pro Leu Thr Lys Gly He Leu Gly Phe Val Phe 50 55 60

Thr Leu Thr Val Pro Ser Glu Arg Gly Leu Gin Arg Arg Arg Phe Val 65 70 75 80

Gin Asn Ala Leu Asn Gly Asn Gly Asp Pro Asn Asn Met Asp Arg Ala 85 90 95

Val Lys Leu Tyr Lys Lys Leu Lys Arg Glu lie Thr Phe His Gly Ala 100 105 110

Lys Glu Val Ser Leu Ser Tyr Ser Thr Gly Ala Leu Ala Ser Cys Met 115 120 125

Gly Leu lie Tyr Asn Arg Met Gly Thr Val Thr Thr Glu Ala Ala Phe 130 135 140

Gly Leu Val Cys Ala Thr Cys Glu Gin lie Ala Asp Ser Gin His Arg 145 150 155 160

Ser His Arg Gin Met Ala Thr Thr Thr Asn Pro Leu lie Arg His Glu 165 170 175

Asn Arg Met Val Leu Ala Ser Thr Thr Ala Lys Ala Met Glu Gin Met 180 185 190

Ala Gly Ser Ser Glu Gin Pro Ala Glu Ala Met Glu Val Ala Asn Gin 195 200 205

Thr Arg Gin Met Val His Ala Met Arg Thr lie Gly Thr His Pro Ser 210 215 220

Ser Ser Ala Gly Leu Lys Asp Asp Leu Leu Glu Asn Leu Gin Ala Tyr G, „ Lys Arg Met Gly Val Gln Met Gin Phe Lys

&lt;210&gt; 46 &lt;211&gt; 96 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 46

Met Ser Uu Leu Thr Glu Val Glu Thr Pro Thr Arg Ser Glu Trp Glu 15 10 15

CysA^CysSerAspSerSerProLeuVa, He Ala Ala Asn I,e He

Gly lie Leu His Leu lie Leu Trp lie Thr Asp Arg Leu Phe Phe Lys 35 40 45

Cys lie Tyr Arg Arg Phe Lys Tyr Gly Leu Lys Arg Gly Pro Ser Thr -49- 159890 - Sequence Listing.doc 201249996 50 55 60

Glu Gly Val Pro Glu Ser Met Arg Glublu Tyr Gin Gin Glu Gin Gin 65 70 75 80

Ser Ala Val Asp Val Asp Asp Gly His Phe Va] Asn lie Glu Leu Glu 85 90 95

&lt;210&gt; 47 &lt;211&gt; 252 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt;

Met Ser Leu Leu Thr Glu Val Glu Thr Tyr Val Leu Ser He lie Pro 15 10 15

Ser Gly Pro Leu Lys Ala Glu lie Ala Gin Arg Leu Glu Ser Val Phe 20 25 30

Ala Gly Lys Asn Thr Asp Leu Glu Ala Leu Met Glu Trp Leu Lys Thr 35 40 45

Arg Pro lie Leu Ser Pro Leu Thr Lys Gly He Leu Gly Phe Val Phe 50 55 60

Thr Leu Thr Val Pro Ser Glu Arg Gly Leu Gin Arg Arg Arg Phe Val 65 70 75 80

Gin Asn Ala Leu Asn Gly Asn Gly Asp Pro Asn Asn Met Asp Arg Ala 85 90 95

Val Lys Leu Tyr Lys Lys Leu Lys Arg Glu lie Thr Phe His Gly Ala 100 105 110

Lys Glu Val Ser Leu Ser Tyr Ser Thr Gly Ala Leu Ala Ser Cys Met 115 120 125

Gly Leu lie Tyr Asn Arg Met Gly Thr Val Thr Thr Glu Ala Ala Phe 130 135 140

Gly Leu Val Cys Ala Thr Cys Glu Gin lie Ala Asp Ser Gin His Arg 145 150 155 160

Ser His Arg Gin Met Ala Thr Thr Thr Asn Pro Leu lie Arg His Glu 165 170 175

Asn Arg Met Val Leu Ala Ser Thr Thr Ala Lys Ala Met Glu Gin Met 180 185 190

Ala Gly Ser Ser Glu Gin Pro Ala Glu Ala Met Glu Val Ala Asn Gin 195 200 205

Thr Arg Gin Met Val His Ala Met Arg Thr He Gly Thr His Pro Ser 210 215 220

Ser Ser Ala Gly Leu Lys Asp Asp Leu Leu Glu Asn Leu Gin Ala Tyr -50· 159890 · Sequence Listing.doc 201249996 225 230 235 240

Gin Lys Arg Met Gly Val Gin Met Gin Arg Phe Lys 245 250

&lt;210&gt; 48 &lt;211&gt; 96 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 48

Met Ser Leu Leu Thr Glu Val Glu Thr Pro Thr Arg Ser Glu Trp Glu 15 10 15

Cys Arg Cys Ser Asp Ser Ser Pro Leu Val lie Ala Ala Asn He lie 20 25 30

Gly lie Leu His Leu lie Leu Trp lie Thr Asp Arg Leu Phe Phe Lys 35 40 45

Cys lie Tyr Arg Arg Phe Lys Tyr Gly Leu Lys Arg Gly Pro Ser Thr 50 55 60

Glu Gly Val Pro Glu Ser Met Arg Glu Glu Tyr Gin Gin Glu Gin Gin 65 70 75 80

Ser Ala Val Asp Val Asp Asp Gly His Phe Val Asn lie Glu Leu Glu 85 90 95

&lt;210&gt; . 49 &lt;211&gt; 252 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt;

Met Ser Leu Leu Thr Glu Val Glu Thr Tyr Val Leu Ser lie lie Pro 15 10 15

Ser Gly Pro Leu Lys Ala Glu lie Ala Gin Arg Leu Glu Ser Val Phe 20 25 30

Ala Gly Lys Asn Thr Asp Leu Glu Ala Leu Met Glu Trp Leu Lys Thr 35 40 45

Arg Pro lie Leu Ser Pro Leu Thr Lys Gly lie Leu Gly Phe Val Phe 50 55 60

Thr Leu Thr Val Pro Ser Glu Arg Gly Leu Gin Arg Arg Arg Phe Val 65 70 75 80

Gin Asn Ala Leu Asn Gly Asn Gly Asp Pro Asn Asn Met Asp Arg Ala 85 90 95

Val Lys Leu Tyr Lys Lys Leu Lys Arg Glu lie Thr Phe His Gly Ala 100 105 110

Lys Glu Val Ser Leu Ser Tyr Ser Thr Gly Ala Leu Ala Ser Cys Met 115 120 125 -51 - 159890 - Sequence Listing.doc 201249996

Gly Leu lie Tyr Asn Arg Met Gly Thr Val Thr Thr Glu Ala Ala Phe 130 135 140

Gly Leu Val Cys Ala Thr Cys Glu Gin lie Ala Asp Ser Gin His Arg 145 150 155 160

Ser His Arg Gin Met Ala Thr Thr Thr Asn Pro Leu lie Arg His Glu 165 170 175

Asn Arg Met Val Leu Ala Ser Thr Thr Ala Lys Ala Met Glu Gin Met 180 185 190

Ala Gly Ser Ser Glu Gin Pro Ala Glu Ala Met Glu Val Ala Asn Gin 195 200 205

Thr Arg Gin Met Val His Ala Met Arg Thr lie Gly Thr His Pro Ser 210 215 220

Ser Ser Ala Gly Leu Lys Asp Asp Leu Leu Glu Asn Leu Gin Ala Tyr 225 230 235 240

Gin Lys Arg Met Gly Val Gin Met Gin Arg Phe Lys 245 250

&lt;210&gt; 50 &lt;211&gt; 96 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 50

Met Ser Leu Leu Thr Glu Val Glu Thr Pro Thr Arg Ser Glu Trp Glu 15 10 15

Cys Arg Cys Ser Asp Ser Ser Pro Leu Val lie Ala Ala Asn lie lie 20 25 30

Gly lie Leu His Leu lie Leu Trp lie Thr Asp Arg Leu Phe Phe Lys 35 40 45

Cys lie Tyr Arg Arg Phe Lys Tyr Gly Leu Lys Arg Gly Pro Ser Thr 50 55 60

Glu Gly Val Pro Glu Ser Met Arg Glu Glu Tyr Gin Gin Glu Gin Gin 65 70 75 80

Ser Ala Val Asp Val Asp Asp Gly His Phe Val Asn lie Glu Leu Glu 85 90 95

&lt;210&gt; 51 &lt;211&gt; 252 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt;

Met Ser Leu Leu Thr Glu Val Glu Thr Tyr Val Leu Ser lie lie Pro 15 10 15 -52- 159890 - Sequence Listing.doc 201249996

Ser Gly Pro Leu Lys Ala Glu He Ala Gin Arg 20 25

Leu Glu Ser Val Phe 30

Ala Gly Lys Asn Thr Asp Leu Glu Ala Leu Met 35 40

Glu Trp Leu Lys Thr 45

Arg Pro lie Leu Ser Pro Leu Tlir Lys Gly lie 50 55

Leu Gly Phe Val Phe 60

Thr Leu Thr Val Pro Ser Glu Arg Gly Leu Gin 65 70 75

Arg Arg Arg Phe Val 80

Gin Asn Ala Leu Asn Gly Asn Gly Asp Pro Asn 85 90

Asn Met Asp Arg Ala 95

Val Lys Leu Tyr Lys Lys Lyu Lys Arg Glu lie 100 105

Thr Phe His Gly Ala 110

Lys Glu Val Ser Leu Ser Tyr Ser Thr Gly Ala 115 120

Leu Ala Ser Cys Met 125

Gly Leu lie Tyr Asn Arg Met Gly Tlir Val Thr 130 135

Thr Glu Ala Ala Phe 140

Gly Leu Val Cys Ala Thr Cys Glu Gin lie Ala 145 150 155

Asp Ser Gin His Arg 160

Ser His Arg Gin Met Ala Thr Thr Thr Asn Pro 165 170

Leu lie Arg His Glu 175

Asn Arg Met Val Leu Ala Ser Thr Thr Ala Lys 180 185

Ala Met Glu Gin Met 190

Ala Gly Ser Ser Glu Gin Ala Ala Glu Ala Met 195 200

Glu Val Ala Asn Gin 205

Thr Arg Gin Met Val His Ala Met Arg Thr lie 210 215

Gly Thr His Pro Ser 220

Ser Ser Ala Gly Leu Lys Asp Asp Leu Leu Glu 225 230 235

Asn Leu Gin Ala Tyr 240

Gin Lys Arg Met Gly Val Gin Met Gin Arg Phe 245 250

Lys &lt;210&gt; 52 &lt;211&gt; 97 &lt;212&gt; PRT &lt;213&gt; H1N1 swine influenza virus &lt;400&gt; 52 Met Ser Leu Leu Thr Glu Val Glu Thr Pro Thr 1 5 10

Arg Ser Glu Trp Glu 15

Cys Arg Cys Ser Asp Ser Ser Asp Pro Leu Val 20 25

He Ala Ala Asn lie 30 159890 - Sequence Listing doc -53· 201249996 lie Gly lie Leu His Leu lie Leu Trp lie Thr Asp Arg Leu Phe Phe 35 40 45

Lys Cys lie Tyr Arg Arg Phe Lys Tyr Gly Leu Lys Arg Gly Pro Ser 50 55 60

Thr Glu Gly Val Pro Glu Ser Met Arg Glu Glu Tyr Gin Gin Glu Gin 65 70 75 80

Gin Ser Ala Val Asp Val Asp Asp Gly His Phe Val Asn lie Glu Leu 85 90 95

Glu -54· 159890· Sequence Listing.doc

Claims (1)

201249996 VII. Patent application scope: 1. An influenza virus vaccine comprising an influenza virus having at least the following: a) a hemagglutinin (HA) gene segment comprising a derivative derived from pandemic influenza A The amino acid sequence of the H1/N1 influenza virus 1 isolate SEQ ID NO: 34, and b) the neuraminidase (ΝΑ) gene segment, which is derived from the prevalence of pandemic influenza A Η 1 /Ν1 The amino acid sequence of the Viral Virus 1 isolate is SEQ ID NO: 39. 2. The vaccine of claim 1, wherein the HA gene segment comprises an amino acid sequence encoded by the nucleotide sequence of segment 4 of virus 1 (SEQ ID NO: 4). 3. The vaccine of claim 1, wherein the HA gene segment comprises a branched 7-specific HA sequence of pandemic influenza A H1/N1 influenza virus. 4. The vaccine of claim 1, wherein the HA gene segment comprises a mutation in the D222 residue in the wild type influenza AH1/N1 influenza virus. 5. The vaccine of claim 4, wherein the HA gene segment comprises a D222E mutation. 6. The vaccine of claim 4, wherein the HA gene segment comprises a S203T mutation relative to a wild-type influenza A H1/N1 influenza virus. 7. The vaccine of claim 4, wherein the HA gene segment further comprises a P83S mutation, an I321V mutation; a P83S mutation and an I321V mutation. 8. The vaccine of claim 1, wherein the NA gene segment comprises an amino acid sequence encoded by the nucleotide sequence of segment 6 of virus 1 (SEQ ID NO: 6). 9. The vaccine of claim 1, wherein the NA gene segment is greater than the wild type 159890.doc 201249996 The influenza A H1/N1 influenza virus comprises a Y275H mutation. 10. The vaccine of claim 9, wherein the NA gene segment further comprises an I106V, D248N mutation; a li〇6V mutation and a D248N mutation. 11. The vaccine of claim 1, further comprising: c) a matrix (MA) A gene segment encoding an M1 peptide comprising the amino acid sequence SEQ ID NO: 50 derived from the virus 1 isolate of the pandemic influenza A H1/N1 influenza virus. 12. A vaccine according to claim π, wherein the VIII gene segment comprises an amino acid sequence encoded by the nucleotide sequence of segment 7 of virus 1 (SEq id NO: 7). 13. The vaccine of claim 11, wherein the matrix (MA) gene segment further encodes an M2 peptide comprising an amino acid derived from a virus 1 isolate of pandemic influenza a H1/N1 influenza virus Sequence SEq ID NO: 51 〇 14. A vaccine according to claim η, wherein the μ-like shape comprises a Α198Ρ mutation. 1 5. The use of the influenza vaccine according to any one of claims 1 to 14, which is for the manufacture of a medicament for immunizing a patient against an influenza virus. 16. Use of an influenza vaccine according to any one of claims 1 to 14 for the manufacture of a medicament for immunizing a patient against a hypervirulent influenza virus. 17. Types of claims i to 14 The use of any of the influenza vaccines for the manufacture of a medicament for immunizing a patient against oseltamivir-resistant influenza virus. The vaccine of any one of claims 1 to 14, wherein the vaccine is a redistribution vaccine. 159890.doc 201249996 19. The vaccine of claim 18, wherein the redistributive influenza virus is obtained by a conventional redistribution method. The vaccine of any one of claims 1 to 14, wherein the vaccine is a live attenuated vaccine or a non-activated vaccine. 21. The vaccine of claim 20, wherein the vaccine is formulated for oral or intranasal administration. 22. A method of preparing a vaccine for immunizing an individual with a hypervirulent influenza virus strain, which comprises mixing a redistributive influenza virus and a carrier as claimed in claim 14 and optionally using an adjuvant The steps. 159890.doc