MODIFIED HUMAN PAPILLOMAVIRUS TYPE 52 L1 PROTEIN AND USE THEREOF

Abstract

The present application relates to a modified human papillomavirus (HPV) type 52 L1 protein and a use thereof. Specifically, the present application relates to a HPV type 52 L1 protein, a nucleotide encoded thereby, a carrier comprising the nucleotide, a cell comprising the carrier, a pentamer or virus-like particle consisting of the HPV-52 L1 protein, a vaccine comprising the pentamer or virus-like particle and a vaccine adjuvant, and a use thereof in the prevention of HPV infections and HPV infection-related diseases.

Claims

1. A modified HPV52 L1 protein comprising a modification selected from the group consisting of the following or a combination thereof, when compared with a wild-type HPV52 L1 protein: mutating the amino acid at position 447 from aspartate to glutamate; deleting 1 to 20 successive or nonsuccessive amino acids at the N-terminus; deleting 1 to 25 successive or nonsuccessive amino acids at the C-terminus; substituting one or more amino acids at positions 1 to 20 at the N-terminus; and substituting one or more amino acids at positions 1 to 25 at the C-terminus.

2. The modified HPV52 L1 protein according to claim 1, which is as shown in the sequence selected from SEQ ID No. 2 to SEQ ID No. 29.

3. A polynucleotide encoding the modified HPV52 L1 protein according to claim 1, wherein the sequence of the polynucleotide is whole-gene optimized using insect cell codons.

4. The polynucleotide according to claim 3, which is as shown in the sequence selected from SEQ ID No. 31 to SEQ ID No. 58.

5. A vector comprising the polynucleotide according to claim 3, wherein the vector is selected from the group consisting of plasmid, recombinant Bacmid, and recombinant baculovirus.

6. A host cell comprising the vector according to claim 5, wherein the host cell is selected from the group consisting of E. coli, yeast cell and insect cell.

7. A multimer wherein: the multimer is a pentamer or a virus-like particle; the multimer comprises the modified HPV52 L1 protein according to claim 1, or is formed by the modified HPV52 L1 protein according to claim 1.

8. A vaccine for the prevention of papillomavirus infection or related diseases, said vaccine comprises: the multimer according to claim 7, an adjuvant, and an excipient or carrier for vaccines; wherein the adjuvant is an adjuvant for human use.

9. (canceled)

10. (canceled)

11. The modified HPV52 L1 protein according to claim 1, wherein the wild-type HPV52 L1 protein is as shown in the sequence selected from the group consisting of NCBI Accession No. AEI61557.1, ABU55797.1, AEI61589.1, AIF71344.1, APQ44868.1, AEI61581.1, AIF71350.1, and CAD1814034.1.

12. The modified HPV52 L1 protein according to claim 1, wherein the wild-type HPV52 L1 protein is as shown in SEQ ID No. 1.

13. The modified HPV52 L1 protein according to claim 1, deleting 2, 4, 5, 8, 10, 13, 14, 15, 18, or 20 successive or nonsuccessive amino acids at the N-terminus.

14. The modified HPV52 L1 protein according to claim 1, deleting 13 amino acids at the N-terminus and substituting with any one selected from the group consisting of serine, serine-glutamate, serine-glutamate-arginine, and proline-serine-glutamate-alanine-threonine.

15. The modified HPV52 L1 protein according to claim 1, deleting 19 or 25 successive or nonsuccessive amino acids at the C-terminus.

16. The modified HPV52 L1 protein according to claim 1, substituting one or more basic amino acids at positions 1 to 23 at the C-terminus with any one selected from the group consisting of polar uncharged amino acid, non-polar amino acid, and acidic amino acid; wherein the basic amino acid is selected from arginine and/or lysine; wherein the polar uncharged amino acid is selected from the group consisting of glycine, serine, and threonine; wherein the non-polar amino acid is selected from alanine and/or valine; and wherein the acidic amino acid is selected from aspartate and/or glutamate.

17. The vaccine according to claim 8, further comprising one of the following or a combination thereof: mucosa-tropic HPV virus-like particle or chimeric virus-like particle, skin-tropic HPV virus-like particle or chimeric virus-like particle.

Description

DESCRIPTION OF THE DRAWINGS

[0050] FIG. 1A and FIG. 1B show the expression identification of the wild-type HPV52 L1 and 28 mutants thereof in Example 4 of the present application in insect cells. The results show that the wild-type HPV52 L1 and 28 mutants thereof can all be expressed in insect cells. Lanes 1 to 15 of FIG. 1A represent wild-type HPV52L1, 52L1D447EC19, 52L1N2, 52L1N4, 52L1N5, 52L1N8, 52L1N10, 52L1N13, 52L1N15, 52L1N18, 52L1N20, 52L1CS1, 52L1CS2, 52L1CS3 and 52L1CS4, respectively; Lanes 1 to 14 of FIG. 2A represent 52L1CS5, 52L1CS6, 52L1CS7, 52L1CS8, 52L1CS9, 52L1N13CS1, 52L1N13CS2, 52L1N13CS3, 52L1NS1C19, 52L1NS1C25, 52L1NS2C19, 52L1NS3C19, 52L1NS4C19 and 52L1N14C25, respectively.

[0051] FIGS. 2A to 2K show the dynamic light scattering analysis results of the wild-type HPV52L1, 52L1D447EC19, 52L1N13, 52L1CS5, 52L1CS6, 52L1CS7, 52L1CS9, 52L1N13CS1, 52L1N13CS2, 52L1NS3C19 and 52L1NS4C19 mutant proteins obtained after purification in Example 5 of the present application. The results show that the hydraulic diameters of virus-like particles formed by wild-type HPV52L1, 52L1D447EC19, 52L1N13, 52L1CS5, 52L1CS6, 52L1CS7, 52L1CS9, 52L1N13CS1 and 52L1N13CS2 recombinant proteins are 123.1 nm, 104.9 nm, 71.56 nm, 108.9 nm, 130.4 nm, 116 nm, 124 nm, 111.9 nm, 127.2 nm and 129.9 nm, respectively, and the percentage of particle assembly is 100%. 52L1NS3C19 is not assembled. FIG. 2A represents wild-type HPV52L1; FIG. 2B represents 52L1D447EC19; FIG. 2C represents 52L1N13; FIG. 2D represents 52L1CS5; FIG. 2E represents 52L1CS6; FIG. 2F represents 52L1CS7; FIG. 2G represents 52L1CS9; FIG. 2H represents 52L1N13CS1; FIG. 2I represents 52L1N13CS2; FIG. 2J represents 52L1NS3C19; FIG. 2K represents 52L1NS4C19.

[0052] FIG. 3A to FIG. 3I show the transmission electron microscopy observation results of the wild-type HPV52 L1, 52L1D447EC19, 52L1CS5, 52L1CS6, 52L1CS7, 52L1CS9, 52L1N13CS1, 52L1N13CS2 and 52L1NS4C19 VLPs obtained after purification in Example 6 of the present application. A large number of virus-like particles with diameters of about 40-55 nm can be seen in the field. The particle size is consistent with the theoretical value and has good uniformity. Bar=100 nm. FIG. 3A represents wild-type HPV52L1; FIG. 3B represents 52L1D447EC19; FIG. 3C represents 52L1CS5; FIG. 3D represents 52L1CS6; FIG. 3E represents 52L1CS7; FIG. 3F represents 52L1CS9; FIG. 3G represents 52L1N13CS1; FIG. 3H represents 52L1N13CS2; FIG. 3I represents 52L1NS4C19.

[0053] FIG. 4 shows the analysis of HPV52 neutralizing antibody titers in immune serum after inoculating mice with wild-type HPV52L1, 52L1D447EC19, 52L1N13, 52L1CS5, 52L1CS6, 52L1CS7, 52L1CS9, 52L1N13CS1, 52L1N13CS2, 52L1NS3C19 and 52L1NS4C19 VLPs in Example 7 of the present application. ***: P<0.001.

DETAILED DESCRIPTION OF THE INVENTION

[0054] The present application will be further illustrated by the non-limiting examples below. It is well known to those skilled in the art that many modifications can be made to the present application without departing from the spirit of the present application, and such modifications also fall within the scope of the present application. The following embodiments are only used to illustrate the present application and should not be regarded as limiting the scope of the present application, as the embodiments are necessarily diverse. The terms used in the present specification are intended only to describe particular embodiments but not as limitations. The scope of the present application has been defined in the appended claims.

[0055] Unless otherwise specified, all the technical and scientific terms used in the present specification have the same meaning as those generally understood by those skilled in the technical field to which the present application relates. Preferred methods and materials of the present application are described below, but any method and material similar or equivalent to the methods and materials described in the present specification can be used to implement or test the present application. Unless otherwise specified, the following experimental methods are conventional methods or methods described in product specifications. Unless otherwise specified, the experimental materials used are easily available from commercial companies. All published literatures referred to in the present specification are incorporated here by reference to reveal and illustrate the methods and/or materials in the published literatures.

Example 1: Synthesis of the Gene of Mutated L1 Protein and Construction of Expression Vectors

[0056] The 28 mutated L1 proteins were as follows respectively:

[0057] 1) 52L1D447EC19: The template was full-length HPV52 L1 gene (the sequence was as shown in SEQ ID NO. 1), and its corresponding amino acid sequence was the sequence No. AEI61557.1 in NCBI database (the sequence was as shown in SEQ ID NO. 30). The polynucleotide sequence encoding HPV52 L1D447EC19 was optimized by insect codons, constructed by deleting the nucleotides 1453-1509 of the HPV52 L1 gene backbone for insect cell codon optimization and mutating the nucleotides 1339-1341 from GAC to GAG (the amino acid sequence was as shown in SEQ ID NO. 2, and the nucleotide sequence was as shown in SEQ ID NO. 31), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0058] 2) 52L1N2: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by deleting the nucleotides 4-6 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 3, and the nucleotide sequence was as shown in SEQ ID NO. 32), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0059] 3) 52L1N4: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by deleting the nucleotides 4-12 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 4, and the nucleotide sequence was as shown in SEQ ID NO. 33), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0060] 4) 52L1N5: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by deleting the nucleotides 4-15 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 5, and the nucleotide sequence was as shown in SEQ ID NO. 34), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0061] 5) 52L1N8: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by deleting the nucleotides 4-24 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 6, and the nucleotide sequence was as shown in SEQ ID NO. 35), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0062] 6) 52L1N10: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by deleting the nucleotides 4-30 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 7, and the nucleotide sequence was as shown in SEQ ID NO. 36), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0063] 7) 52L1N13: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by deleting the nucleotides 4-39 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 8, and the nucleotide sequence was as shown in SEQ ID NO. 37), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0064] 8) 52L1N15: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by deleting the nucleotides 4-45 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 9, and the nucleotide sequence was as shown in SEQ ID NO. 38), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0065] 9) 52L1N18: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by deleting the nucleotides 4-54 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 10, and the nucleotide sequence was as shown in SEQ ID NO. 39), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0066] 10) 52L1N20: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by deleting the nucleotides 4-60 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 11, and the nucleotide sequence was as shown in SEQ ID NO. 40), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0067] 11) 52L1CS1: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by mutating the nucleotides 1447-1449 of HPV52 L1D447EC19 from AAA to GGA and inserting the nucleotide sequence AAAGGTCCTGCATCGAGCGCTCCTAGAACGTCGACGGACGGCTCGGGAGTGGG ACGC after the nucleotide 1452 (the amino acid sequence was as shown in SEQ ID NO. 12, and the nucleotide sequence was as shown in SEQ ID NO. 41), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0068] 12) 52L1CS2: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by mutating the nucleotides 1447-1449 of HPV52 L1D447EC19 from AAA to GGA and inserting the nucleotide sequence AAAGGTCCTGCATCGAGCGCTCCTAGAACGTCGACGGACGGCTCGGGAGTGGA CGGC after the nucleotide 1452 (the amino acid sequence was as shown in SEQ ID NO. 13, and the nucleotide sequence was as shown in SEQ ID NO. 42), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0069] 13) 52L1CS3: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by mutating the nucleotides 1447-1449 of HPV52 L1D447EC19 from AAA to GGA and inserting the nucleotide sequence GGATCGCCTGCATCGAGCGCTCCTAGAACGTCGACGGACGGCTCGGGAGTGAA ACGC after the nucleotide 1452 (the amino acid sequence was as shown in SEQ ID NO. 14, and the nucleotide sequence was as shown in SEQ ID NO. 43), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0070] 14) 52L1CS4: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by mutating the nucleotides 1447-1449 of HPV52 L1D447EC19 from AAA to GGA and inserting the nucleotide sequence GGATCGCCTGCATCGAGCGCTCCTAGAACGTCGACGGACGGCTCGGGAGTGGA CCGC after the nucleotide 1452 (the amino acid sequence was as shown in SEQ ID NO. 15, and the nucleotide sequence was as shown in SEQ ID NO. 44), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0071] 52L1CS5: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by inserting the nucleotide sequence GCTGGTCCTGCCTCTTCCGCACCCGCGACTTCAACCGCTGCCGGCGGAGTTGGG TCG after the nucleotide 1452 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 16, and the nucleotide sequence was as shown in SEQ ID NO. 45), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0072] 16) 52L1CS6: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by inserting the nucleotide sequence GAAGCTCCTGCCTCTTCCGCACCCGGTACTTCAACCGGCTCGAAAGCGGTTGCT GGA after the nucleotide 1452 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 17, and the nucleotide sequence was as shown in SEQ ID NO. 46), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0073] 17) 52L1CS7: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by inserting the nucleotide sequence GCTGGTCCTGCTTCCTCAGCTCCAGCTACCTCAACCGACGGTTCTGGTGTGAAG CGC after the nucleotide 1452 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 18, and the nucleotide sequence was as shown in SEQ ID NO. 47), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0074] 18) 52L1CS8: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by inserting the nucleotide sequence GCTGGTCCTGCTTCCTCAGCTCCACGTACCTCAACCGACGGTTCTGGTGTGAAG CGC after the nucleotide 1452 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 19, and the nucleotide sequence was as shown in SEQ ID NO. 48), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0075] 19) 52L1CS9: The template was HPV52 L1D447EC19 gene (the sequence was as shown in SEQ ID NO. 30). It was constructed by mutating the nucleotides 1441-1443 of HPV52 L1D447EC19 from AGA to GGT, mutating the nucleotides 1447-1449 from AAA to GGC and inserting the nucleotide sequence TCGGGTCCTGCCTCGAGCGCCCCTAGAACGTCGACGGG TGGCTCGGCCGTGGGTAGC after the nucleotide 1452 of HPV52 L1D447EC19 (the amino acid sequence was as shown in SEQ ID NO. 20, and the nucleotide sequence was as shown in SEQ ID NO. 49), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0076] 20) 52L1N13CS1: The template was HPV52 L1N13 gene (the sequence was as shown in SEQ ID NO. 37). It was constructed by mutating the nucleotides 1411-1416 of HPV52 L1N13 from AAACTG to GGCTTG and inserting the nucleotide sequence TCGGGTCCTGCCTCGAGCGCCCCTAGAACGTCGACGGGTGGCTCGGCCGTGGGT AGC after the nucleotide 1416 (the amino acid sequence was as shown in SEQ ID NO. 21, and the nucleotide sequence was as shown in SEQ ID NO. 50), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0077] 21) 52L1N13CS2: The template was HPV52 L1N13 gene (the sequence was as shown in SEQ ID NO. 37). It was constructed by mutating the nucleotides 1405-1407 of HPV52 L1N13 from AGA to GGT, mutating the nucleotides 1411-1416 from AAACTG to GGCTTG and inserting the nucleotide sequence TCGGGTCCTGCCTCGAGCGCCCCTAGAACGTCGACGGGTGGCTCGGCC GTGGGTAGC after the nucleotide 1416 (the amino acid sequence was as shown in SEQ ID NO. 22, and the nucleotide sequence was as shown in SEQ ID NO. 51), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0078] 22) 52L1N13CS3: The template was HPV52 L1N13 gene (the sequence was as shown in SEQ ID NO. 37). It was constructed by inserting the nucleotide sequence GCCGGTCCTGCCTCGAGCGCCCCTGCCACGTCGACGGCTGCGGGAGGCGTGGG TAGC after the nucleotide 1416 of HPV52 L1N13 (the amino acid sequence was as shown in SEQ ID NO. 23, and the nucleotide sequence was as shown in SEQ ID NO. 52), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0079] 23) 52L1NS1C19: The template was HPV52 L1N13 gene (the sequence was as shown in SEQ ID NO. 37). It was constructed by inserting the nucleotide sequence CCTAGCGAGGCTACC between the nucleotides 3/4 of HPV52 L1N13 (the amino acid sequence was as shown in SEQ ID NO. 24, and the nucleotide sequence was as shown in SEQ ID NO. 53), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0080] 24) 52L1NS1C25: The template was HPV52 L1N13 gene (the sequence was as shown in SEQ ID NO. 37). It was constructed by inserting the nucleotide sequence CCTAGCGAGGCTACC between the nucleotides 3/4 of HPV52 L1N13 and deleting the nucleotides 1414-1431 (the amino acid sequence was as shown in SEQ ID NO. 25, and the nucleotide sequence was as shown in SEQ ID NO. 54), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0081] 25) 52L1NS2C19: The template was HPV52 L1N13 gene (the sequence was as shown in SEQ ID NO. 37). It was constructed by inserting the nucleotide sequence TCCGAGCGT between the nucleotides 3/4 of HPV52 L1N13 (the amino acid sequence was as shown in SEQ ID NO. 26, and the nucleotide sequence was as shown in SEQ ID NO. 55), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0082] 26) 52L1NS3C19: The template was HPV52 L1N13 gene (the sequence was as shown in SEQ ID NO. 37). It was constructed by inserting the nucleotide sequence TCCGAG between the nucleotides 3/4 of HPV52 L1N13 (the amino acid sequence was as shown in SEQ ID NO. 27, and the nucleotide sequence was as shown in SEQ ID NO. 56), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0083] 27) 52L1NS4C19: The template was HPV52 L1N13 gene (the sequence was as shown in SEQ ID NO. 37). It was constructed by inserting the nucleotide sequence TCC between the nucleotides 3/4 of HPV52 L1N13 (the amino acid sequence was as shown in SEQ ID NO. 28, and the nucleotide sequence was as shown in SEQ ID NO. 57), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0084] 28) 52L1N14C25: The template was HPV52 L1N13 gene (the sequence was as shown in SEQ ID NO. 37). It was constructed by deleting the nucleotides 4-6 and 1414-1431 of HPV52 L1N13 (the amino acid sequence was as shown in SEQ ID NO. 29, and the nucleotide sequence was as shown in SEQ ID NO. 58), and synthesized by Shanghai Sangon Biotech Co., Ltd.

[0085] The EcoR I/BamH I restriction sites were used to digest the above-mentioned synthesized genes respectively, which were inserted into the commercial expression vector pFastBac1 (produced by Invitrogen) respectively to obtain recombinant expression vectors comprising the HPV52 L1 mutated genes, pFastBac1-52L1D447EC19, pFastBac1-52L1N2, pFastBac1-52L1N4, pFastBac1-52L1N5, pFastBac1-52L1N8, pFastBac1-52L1N10, pFastBac1-52L1N13, pFastBac1-52L1N15, pFastBac1-52L1N18, pFastBac1-52L1N20, pFastBac1-52L1CS1, pFastBac1-52L1CS2, pFastBac1-52L1CS3, pFastBac1-52L1CS4, pFastBac1-52L1CS5, pFastBac1-52L1CS6, pFastBac1-52L1CS7, pFastBac1-52L1CS8, pFastBac1-52L1CS9, pFastBac1-52L1N13CS1, pFastBac1-52L1N13CS2, pFastBac1-52L1N13CS3, pFastBac1-52L1NS 1 C19, pFastBac1-52L1NS1C25, pFastBac1-52L1NS2C19, pFastBac1-52L1NS3C19, pFastBac1-52L1NS4C19 and pFastBac1-52L1N14C25.

[0086] The above-mentioned methods of enzyme digestion, ligation and construction of clones were all well known, for example, the patent CN 101293918 B.

Example 2: Recombinant Bacmid and Recombinant Baculovirus Constructs of the HPV52 L1 Mutant Genes

[0087] The recombinant expression vectors comprising HPV52 L1 mutant gene, pFastBac1-52L1D447EC19, pFastBac1-52L1N2, pFastBac1-52L1N4, pFastBac1-52L1N5, pFastBac1-52L1N8, pFastBac1-52L1N10, pFastBac1-52L1N13, pFastBac1-52L1N15, pFastBac1-52L1N18, pFastBac1-52L1N20, pFastBac1-52L1CS1, pFastBac1-52L1CS2, pFastBac1-52L1CS3, pFastBac1-52L1CS4, pFastBac1-52L1CS5, pFastBac1-52L1CS6, pFastBac1-52L1CS7, pFastBac1-52L1CS8, pFastBac1-52L1CS9, pFastBac1-52L1N13CS1, pFastBac1-52L1N13CS2, pFastBac1-52L1N13C53, pFastBac1-52L1NS 1 C19, pFastBac1-52L1NS1C25, pFastBac1-52L1NS2C19, pFastBac1-52L1NS3C19, pFastBac1-52L1NS4C19 and pFastBac1-52L1N14C25, were used to transform E. coli DH10Bac competent cells respectively, which were screened to obtain recombinant Bacmids. Then the recombinant Bacmids were used to transfect Sf9 insect cells to amplify recombinant baculoviruses within Sf9. Methods of screening of recombinant Bacmid and amplification of recombinant baculovirus were all well known, for example, the patent CN 101148661 B.

Example 3: Expression of HPV52 L1 Mutant Genes in Sf9 Cells

[0088] 519 cells were inoculated with the recombinant baculoviruses of 28 HPV52 L1 mutant genes to express the HPV52 L1 mutant proteins. After incubation at 27 C. for about 80 h, the fermentation broth was collected and centrifuged at 3,000 rpm for 15 min. The supernatant was discarded, and the cells were washed with PBS for use in expression identification and purification. Methods of infection and expression were publicly available, for example, the patent CN 101148661 B.

Example 4: Expression Identification and Comparison of Expression Amounts of HPV52 L1 Mutant Proteins

[0089] 110.sup.6 cells expressing the different HPV52 L1 mutants and wild-type HPV52 L1 described in Example 3 respectively were collected and resuspended in 200 l PBS solution. The cells were sonicated by ultrasonic disruption (Ningbo Scientz Ultrasonic Cell Disruptor, 2 #probe, 100 W, ultrasound 5 s, interval 7 s, total period 3 min) and centrifuged at a high speed of 13,000 rpm for 30 minutes. The lysed supernatant was collected and the total protein concentration in each lysed supernatant was detected by BCA assay. The lysed supernatant was uniformly diluted to 20 ng/l with PBS. 2 l of 6 loading buffer was added to 10 l (i.e., 200 ng) of each diluted lysed supernatant. The samples were denatured at 75 C. for 8 min and subjected to SDS-PAGE electrophoresis and Western blot identification to compare the content of L1 protein (with a size of about 55 kDa) in the lysed supernatant of each mutant. The expression identification results of each mutant L1 protein were as shown in FIG. 1, and the comparison of expression amounts of each mutant L1 protein was as shown in Table 1. Methods of SDS-PAGE electrophoresis and Western blot identification were publicly available, for example, the patent CN 101148661 B.

[0090] Microtiter plates were coated with HPV52L1 monoclonal antibodies prepared by the inventor at 80 ng/well by overnight incubation at 4 C. The plate was blocked with 5% BSA-PBST at room temperature for 2 h and washed 3 times with PBST. The lysed supernatant was subjected to 2-fold serial dilution with PBS. The HPV52L1 VLP standard was also subjected to serial dilution from a concentration of 2 g/ml to 0.0625 g/ml. The diluted samples were added to the plate respectively at 100 l per well and incubated at 37 C. for 1 h. The plate was washed 3 times with PBST, and 1:3000 diluted HPV52L1 rabbit polyclonal antibody was added at 100 l per well and incubated at 37 C. for 1 h. The plate was washed 3 times with PBST, and 1:3000 diluted HRP-labeled goat anti-mouse IgG (1:3000 dilution, ZSGB-Bio Corporation) was added and incubated at 37 C. for 45 minutes. The plate was washed 5 times with PBST, and 100 l of OPD substrate (Sigma) was added to each well for development at 37 C. for 5 minutes. The reaction was stopped with 50 l of 2 M sulfuric acid, and the absorbance at 490 nm was determined. The concentrations of modified HPV52L1 proteins and wild-type HPV52L1 protein in the lysed supernatant were calculated according to the standard curve.

[0091] The results were as shown in Table 1. Different modifications had different effects on the expression level of HPV52L1 protein, among which the expression amount of some modified HPV52L1 proteins was increased, in particular 52L1N13, 52L1CS7, 52L1CS9, 52L1N13CS1, 52L1N13CS2, 52 L1N13CS3, 52 L1NS3C19 and 52 L1NS4C19, all with expression levels above 50 mg/L, much higher than the wild-type HPV52L1 protein.

TABLE-US-00001 TABLE 1 Analysis of expression amounts of HPV52 L1 mutant proteins Expression amount (mg/L) Protein name Batch 1 Batch 2 Batch 3 Average HPV52L1 3 3.5 5 3.83 52L1D447EC19 20 15 21 18.67 52L1N2 5 3 4.5 4.17 52L1N4 14 18 14 15.33 52L1N5 15 11 16 14 52L1N8 15 17 15 15.67 52L1N10 21 20 20 20.33 52L1N13 150 152 155 152.3 52L1N15 4.5 5 4 4.5 52 L1N18 5 4 4 4.33 52 L1N20 16 13 14 14.33 52 L1CS1 25 28 24 25.67 52 L1CS2 31 29 33 31 52 L1CS3 42 38 39 39.67 52 L1CS4 40 44 43 42.33 52 L1CS5 43 40 36 42.33 52 L1CS6 40 35 48 41 52 L1CS7 75 73 79 75.67 52 L1CS8 23 21 20 21.33 52 L1CS9 60 67 63 63.33 52 L1N13CS1 123 109 116 116 52 L1N13CS2 108 112 105 108.33 52 L1N13CS3 74 72 78 74.67 52 L1NS1C19 20 23 25 22.67 52 L1NS1C25 19 16 14 16.33 52 L1NS2C19 10 7 15 10.67 52 L1NS3C19 59 62 62 61 52 L1NS4C19 104 108 99 103.67 52 L1N14C25 40 34 39 37.67

Example 5: Purification and Dynamic Light Scattering Particle Size Analysis of L1 Mutant Proteins

[0092] An appropriate amount of cell fermentation broth of L1 mutants was collected and the cells were resuspended with PBS. PMSF was added to a final concentration of 1 mg/ml. The cells were ultrasonically disrupted (Ningbo Scientz Ultrasonic Cell Disruptor, 2 #probe, 200 W, ultrasound 5 s, interval 7 s, total period 10 min) and centrifuged at 13,000 rpm for 30 min. The supernatant was collected and diluted with PBS to 3-4 mg/mL. Saturated ammonium sulfate solution was added to the supernatant until ammonium sulfate saturation was 30%. The supernatant was let stand for precipitation at 4 C. for 1-2 hours and centrifuged at 13,000 rpm for 30 min. The precipitate was resuspended with an appropriate amount of resuspension buffer (20 mM Na.sub.3PO.sub.4, 50 mM DTT, 300 mM NaCl, pH 6.8) and stored on ice overnight. The chromatography purification step was performed at room temperature. The sample was filtered with 0.45 m filter prior to chromatography and purified sequentially using SP-FF cation exchange chromatography and Q-HP anion exchange chromatography (100 mM NaCl, 20 mM Na.sub.3PO.sub.4, 10 mM DTT, pH 6.8 for loading). The purified product was assembled into VLPs in assembly buffer (500 mM NaCl, 2 mM CaCl.sub.2, 2 mM MgCl.Math.6H.sub.2O, 20 mM HEPES, 0.01% Tween 80, pH 6.0) at 4 C. After 3 days of assembly, it was transferred into stabilization buffer (500 mM NaCl, 10 mM histidine, 0.01% Tween 80, pH 7.2) and stabilized at 4 C. for 2 days. The purification results showed that the purification yield of the modified 52L1 proteins was higher than that of the wild-type 52L1, in particular 52L1N13, 52L1CS7, 52L1CS9, 52L1N13CS1, 52L1N13CS2, 52 L1N13CS3, 52 L1NS3C19 and 52 L1NS4C19, with purification yields above 15 mg/L. The above purification methods were all publicly available, for example, the patents CN 101293918 B, CN 1976718 A, etc.

[0093] The purified protein solutions were subjected to DLS particle size analysis (Zetasizer Nano ZS 90 Dynamic Light Scatterer, Malvern), and the results were as shown in FIG. 2 and Table 2. Except for 52L1N13, the hydraulic diameter of all the rest mutants was above 100 nm, close to that of HPV52L1. The hydraulic diameter of 52L1N13 was only 71.56 nm, possibly suggesting its lower degree of assembly.

TABLE-US-00002 TABLE 2 DLS analysis of HPV52 L1 mutant proteins Protein name Hydraulic diameter (nm) PDI HPV52L1 123.1 0.134 52L1D447EC19 104.9 0.142 52L1N13 71.56 0.141 52L1CS5 108.9 0.126 52L1CS6 130.4 0.111 52L1CS7 116 0.135 52L1CS9 124 0.143 52L1N13CS1 111.9 0.09 52L1N13CS2 127.2 0.139 52L1NS3C19 149.4 0.234 52L1NS4C19 129.9 0.125

Example 6: Transmission Electron Microscopy Observation of HPV52 L1 Mutant VLPs

[0094] HPV52 L1 and mutant proteins thereof were purified and assembled respectively according to the chromatographic purification method described in Example 5. The assembled VLPs were prepared on copper mesh, stained with 1% uranium acetate, fully dried and then observed using JM-1400 electron microscope (Olympus). The transmission electron microscopy images of HPV52 L1, HPV52 L1D447EC19, HPV52 L1CS5, HPV52 L1CS6, HPV52 L1CS7, HPV52 L1CS9, HPV52 L1N13CS1, HPV52 L1N13CS2 and HPV52 L1NS4C19 VLPs were as shown in FIGS. 3A-3I respectively. The diameter of all these mutants was between 40-55 nm. Methods of copper mesh preparation and electron microscopy observation were all publicly available, for example, the patent CN 101148661 B.

Example 7: Immunization of Mice with HPV52 L1 Mutant VLPs and Determination of Neutralizing Antibody Titers

[0095] 4-6 weeks old female BALB/c mice were randomly divided into groups of 5 mice and immunized with 0.1 g VLP by intramuscular injection at Weeks 0, 2 and 4. Tail vein blood was collected 2 weeks after the third immunization and serum was isolated.

[0096] HPV52 pseudovirus was used to detect the neutralizing antibody titers in immune serum, and the results were as shown in Table 3 and FIG. 4. The neutralizing activity of immune serum caused by VLPs produced in insect cell expression systems, such as 52L1D447EC19, 52L1CS5, 52L1CS6, 52L1CS7, 52L1CS9, 52L1N13CS1, 52L1N13CS2 and 52L1NS4C19, was comparable to that caused by HPV52L1, while 52L1N13 immune serum had no neutralizing activity. Methods of pseudovirus preparation and pseudovirus neutralization experiments were all publicly available, for example, the patent CN 104418942A.

TABLE-US-00003 TABLE 3 Neutralizing antibody titers against HPV52 pseudovirus induced by HPV52 L1 mutants in vivo in mice Antigen name Average neutralizing antibody titer HPV52L1 8960 52L1D447EC19 10240 52L1N13 <25 52L1CS5 11520 52L1CS6 8320 52L1CS7 10880 52L1CS9 9600 52L1N13CS1 11520 52L1N13CS2 9600 52L1NS4C19 10880

[0097] In summary, the inventor has found that the mutants obtained by modification of HPV52L1 amino acid sequence have different expression levels. Their degree of assembly and immune activity can both be affected by the mutation modification in an irregular way. Therefore, it cannot be expected that HPV52L1 mutants with high expression level, effective assembly and good immune activity can be obtained by modification of the amino acid sequence. The optimally modified HPV52L1 mutants obtained by screening in the present application can be used in the formulation of multivalent HPV prophylactic vaccine and in the construction of broad-spectrum HPV prophylactic vaccine, and has good research and development prospects.

Description of Sequences

[0098]

TABLE-US-00004 SEQIDNO.1:HPV52L1 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSS GNGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRG QPLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGE HWGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPI DICSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQ GSNSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHINNGICWGNQLFV TVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVM TYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKDYM FWEVDLKEKFSADLDQFPLGRKFLLQAGLQARPKLKRPASSAPRTSTKKKKVKR SEQIDNO.2:52LID447EC19 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSS GNGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRG QPLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGE HWGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPI DICSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQ GSNSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFV TVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVM TYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYM FWEVDLKEKFSADLDQFPLGRKFLLQAGLQARPKL SEQIDNO.3:52L1N2 MVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSG NGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQ PLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEH WGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDI CSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGS NSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTV VDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYI HKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFW EVDLKEKFSADLDQFPLGRKFLLQAGLQARPKL SEQIDNO.4:52L1N4 MRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNG KKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLG VGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWG KGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDENTLQASKSDVPIDICS SVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNS GNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVD TTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHK MDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEV DLKEKFSADLDQFPLGRKFLLQAGLQARPKL SEQIDNO.5:52LIN5 MPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGK KVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGV GISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGK GTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDENTLQASKSDVPIDICSS VCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSG NTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDT TRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHK MDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEV DLKEKFSADLDQFPLGRKFLLQAGLQARPKL SEQIDNO.6:52L1N8 MATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVL VPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGIS GHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTP CNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCK YPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTA TVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRS TNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDA TILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKE KFSADLDQFPLGRKFLLQAGLQARPKL SEQIDNO.7:52LIN10 MVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVP KVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGH PLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCN NNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYP DYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATV QSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTN MTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATIL EDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKF SADLDQFPLGRKFLLQAGLQARPKL SEQIDNO.8:52L1N13 MPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPKVS GLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHPLL NKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNNNS GNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYL QMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSS AFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTL CAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILED WQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKFSA DLDQFPLGRKFLLQAGLQARPKL SEQIDNO.9:52L1N15 MVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPKVSGL QYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHPLLNK FDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNNNSGN PGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYLQM ASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSSAFF PTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTLCAE VKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILEDWQF GLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKFSADLD QFPLGRKFLLQAGLQARPKL SEQIDNO.10:52L1N18 MSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPKVSGLQYR VFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHPLLNKFDD TETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNNNSGNPGD CPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYLQMASE PYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSSAFFPTP SGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTLCAEVK KESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILEDWQFGLT PPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKFSADLDQFPL GRKFLLQAGLQARPKL SEQIDNO.11:52L1N20 MVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPKVSGLQYRVF RIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHPLLNKFDDTE TSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNNNSGNPGDCP PLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYLQMASEPY GDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSSAFFPTPSG SMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTLCAEVKKE STYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILEDWQFGLTPP PSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKFSADLDQFPLG RKFLLQAGLQARPKL SEQIDNO.12:52L1CS1 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSS GNGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRG QPLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGE HWGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDENTLQASKSDVPI DICSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQ GSNSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFV TVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVM TYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYM FWEVDLKEKFSADLDQFPLGRKFLLQAGLQARPGLKGPASSAPRTSTDGSGVGR SEQIDNO.13:52L1CS2 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSS GNGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRG QPLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGE HWGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPI DICSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQ GSNSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFV TVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVM TYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYM FWEVDLKEKFSADLDQFPLGRKFLLQAGLQARPGLKGPASSAPRTSTDGSGVDG SEQIDNO.14:52L1CS3 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSS GNGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRG QPLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGE HWGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPI DICSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQ GSNSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFV TVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVM TYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYM FWEVDLKEKFSADLDQFPLGRKFLLQAGLQARPGLGSPASSAPRTSTDGSGVKR SEQIDNO.15:52L1CS4 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSS GNGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRG QPLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGE HWGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPI DICSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQ GSNSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFV TVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVM TYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYM FWEVDLKEKFSADLDQFPLGRKFLLQAGLQARPGLGSPASSAPRTSTDGSGVDR SEQIDNO.16:52L1CS5 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSS GNGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRG QPLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGE HWGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDENTLQASKSDVPI DICSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQ GSNSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFV TVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVM TYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYM FWEVDLKEKFSADLDQFPLGRKFLLQAGLQARPKLAGPASSAPATSTAAGGVGS SEQIDNO.17:52L1CS6 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSS GNGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRG QPLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGE HWGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPI DICSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQ GSNSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFV TVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVM TYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYM FWEVDLKEKFSADLDQFPLGRKFLLQAGLQARPKLEAPASSAPGTSTGSKAVAG SEQIDNO.18:52L1CS7 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSS GNGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRG QPLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGE HWGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPI DICSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQ GSNSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHINNGICWGNQLFV TVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVM TYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYM FWEVDLKEKFSADLDQFPLGRKFLLQAGLQARPKLAGPASSAPATSTDGSGVKR SEQIDNO.19:52L1CS8 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSS GNGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRG QPLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGE HWGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDENTLQASKSDVPI DICSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQ GSNSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFV TVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVM TYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYM FWEVDLKEKFSADLDQFPLGRKFLLQAGLQARPKLAGPASSAPRTSTDGSGVKR SEQIDNO.20:52L1CS9 MSVWRPSEATVYLPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSS GNGKKVLVPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRG QPLGVGISGHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGE HWGKGTPCNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDENTLQASKSDVPI DICSSVCKYPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQ GSNSGNTATVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFV TVVDTTRSTNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVM TYIHKMDATILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYM FWEVDLKEKFSADLDQFPLGRKFLLQAGLQAGPGLSGPASSAPRTSTGGSAVGS SEQIDNO.21:52L1N13CS1 MPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPKVS GLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHPLL NKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNNNS GNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYL QMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSS AFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTL CAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILED WQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKFSA DLDQFPLGRKFLLQAGLQARPGLSGPASSAP481RTSTGGSAVGS SEQIDNO.22:52L1N13CS2 MPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPKVS GLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHPLL NKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNNNS GNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYL QMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSS AFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTL CAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILED WQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKFSA DLDQFPLGRKFLLQAGLQAGPGLSGPASSAP481RTSTGGSAVGS SEQIDNO.23:52L1N13CS3 MPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPKVS GLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHPLL NKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNNNS GNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYL QMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSS AFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTL CAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILED WQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKFSA DLDQFPLGRKFLLQAGLQARPKLAGPASSAP481ATSTAAGGVGS SEQIDNO.24:52L1NS1C19 MPSEATPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVL VPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGIS GHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTP CNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCK YPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTA TVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRS TNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDA TILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKE KFSADLDQFPLGRKFLLQAGLQARPKL SEQIDNO.25:52L1NS1C25 MPSEATPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVL VPKVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGIS GHPLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTP CNNNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDENTLQASKSDVPIDICSSVCK YPDYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTA TVQSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRS TNMTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDA TILEDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKE KFSADLDQFPLGRKFLLQAGL SEQIDNO.26:52L1NS2C19 MSERPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVP KVSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGH PLLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCN NNSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYP DYLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATV QSSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTN MTLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATIL EDWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKF SADLDQFPLGRKFLLQAGLQARPKL SEQIDNO.27:52L1NS3C19 MSEPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPK VSGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHP LLNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNN NSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPD YLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQ SSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNM TLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILE DWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKFS ADLDQFPLGRKFLLQAGLQARPKL SEQIDNO.28:52L1NS4C19 MSPPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPKV SGLQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHPL LNKFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNN NSGNPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPD YLQMASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQ SSAFFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNM TLCAEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILE DWQFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKFS ADLDQFPLGRKFLLQAGLQARPKL SEQIDNO.29:52L1N14C25 MPVPVSKVVSTDEYVSRTSIYYYAGSSRLLTVGHPYFSIKNTSSGNGKKVLVPKVSG LQYRVFRIKLPDPNKFGFPDTSFYNPETQRLVWACTGLEIGRGQPLGVGISGHPLLN KFDDTETSNKYAGKPGIDNRECLSMDYKQTQLCILGCKPPIGEHWGKGTPCNNNSG NPGDCPPLQLINSVIQDGDMVDTGFGCMDFNTLQASKSDVPIDICSSVCKYPDYLQ MASEPYGDSLFFFLRREQMFVRHFFNRAGTLGDPVPGDLYIQGSNSGNTATVQSSA FFPTPSGSMVTSESQLFNKPYWLQRAQGHNNGICWGNQLFVTVVDTTRSTNMTLC AEVKKESTYKNENFKEYLRHGEEFDLQFIFQLCKITLTADVMTYIHKMDATILEDW QFGLTPPPSASLEDTYRFVTSTAITCQKNTPPKGKEDPLKEYMFWEVDLKEKFSADL DQFPLGRKFLLQAGL SEQIDNO.30:HPV52L1nt ATGTCCGTGTGGCGGCCTAGTGAGGCCACTGTGTACCTGCCTCCTGTACCTGTCT CTAAGGTTGTAAGCACTGATGAGTATGTGTCTCGCACAAGCATCTATTATTATGCA GGCAGTTCTCGATTACTAACAGTAGGACATCCCTATTTTTCTATTAAAAACACCA GTAGTGGTAATGGTAAAAAAGTTTTAGTTCCCAAGGTGTCTGGCCTGCAATACA GGGTATTTAGAATTAAATTGCCGGACCCTAATAAATTTGGTTTTCCGGATACATCT TTTTATAACCCAGAAACCCAAAGGTTGGTGTGGGCCTGTACAGGCTTGGAAATT GGTAGGGGACAGCCTTTAGGTGTGGGTATTAGTGGGCATCCTTTATTAAACAAGT TTGATGATACTGAAACCAGTAACAAATATGCTGGTAAACCTGGTATAGATAATAG AGAATGTTTATCTATGGATTATAAGCAGACTCAGTTATGCATTTTAGGATGCAAAC CTCCTATAGGTGAACATTGGGGTAAGGGAACCCCTTGTAATAATAATTCAGGAAA TCCTGGGGATTGTCCTCCCCTACAACTCATTAACAGTGTAATACAGGATGGGGAC ATGGTAGATACAGGATTTGGTTGCATGGATTTTAATACCTTGCAAGCTAGTAAAA GTGATGTGCCCATTGATATATGTAGCAGTGTATGTAAGTATCCAGATTATTTGCAA ATGGCTAGCGAGCCATATGGTGACAGTTTGTTCTTTTTTCTTAGACGTGAGCAAA TGTTTGTTAGACACTTTTTTAATAGGGCTGGTACCTTAGGTGACCCTGTGCCAGG TGATTTATATATACAAGGGTCTAACTCTGGCAATACTGCCACTGTACAAAGCAGT GCTTTTTTTCCTACTCCTAGTGGTTCTATGGTAACCTCAGAATCCCAATTATTTAAT AAACCGTACTGGTTACAACGTGCGCAGGGCCACAATAATGGCATATGTTGGGGC AATCAGTTGTTTGTCACAGTTGTGGATACCACTCGTAGCACTAACATGACTTTAT GTGCTGAAGTTAAAAAGGAAAGCACATATAAAAATGAAAATTTTAAGGAATACC TTCGTCATGGCGAGGAATTTGATTTACAATTTATTTTTCAATTGTGCAAAATTACA TTAACAGCTGATGTTATGACATACATTCATAAGATGGATGCCACTATTTTAGAGGA CTGGCAATTTGGCCTTACCCCACCACCGTCTGCATCTTTGGAGGACACATACAGA TTTGTAACTTCTACTGCTATAACTTGTCAAAAAAACACACCACCTAAAGGAAAG GAAGATCCTTTAAAGGACTATATGTTTTGGGAGGTGGATTTAAAAGAAAAGTTTT CTGCAGATTTAGATCAGTTTCCTTTAGGTAGGAAGTTTTTGTTACAGGCAGGGCT ACAGGCTAGGCCCAAACTAAAACGCCCTGCATCATCAGCCCCACGTACCTCCAC AAAGAAGAAAAAGGTTAAAAGGTAA SEQIDNO.31:52L1D447EC19nt ATGTCCGTGTGGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTT CTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCT GGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGT CCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACC GCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAG TTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAAT TGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAA GTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAA CCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGC AAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCA GGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGAT GGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTT CCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTA TCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGG GAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCT GTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTG CAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGC CAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGC ATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCA ATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATT TCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCT CTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTA CCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGA AGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCC ACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCT CAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCT CTTGCAAGCAGGACTGCAAGCTAGACCTAAACTGTAA SEQIDNO.32:52L1N2nt ATGGTGTGGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTTCTA AAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCTGGT AGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGTCCT CAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACCGCG TCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAGTTT CTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAATTGG CAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAAGTT CGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAACC GTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGCAA GCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCAGG AAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGATGGT GACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTTCCA AGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTATCT GCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGGGA GCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCTGTC CCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTGCAG TCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGCCAA CTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGCATC TGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCAATA TGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATTTC AAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCTCT GCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTACC ATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGAA GACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCCA CCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCTC AAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCTC TTGCAAGCAGGACTGCAAGCTAGACCTAAACTGTAA SEQIDNO.33:52L1N4nt ATGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTTCTAAAGTGG TCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCTGGTAGTTCA AGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGTCCTCAGGAA ACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACCGCGTCTTCC GTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAGTTTCTATAA CCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAATTGGCAGGG GTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAAGTTCGACG ACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAACCGTGAAT GCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGCAAGCCGC CTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCAGGAAACC CAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGATGGTGACAT GGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTTCCAAGAGC GATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTATCTGCAAAT GGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGGGAGCAGAT GTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCTGTCCCCGGA GACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTGCAGTCTTCC GCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGCCAACTCTTT AATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGCATCTGCTGG GGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCAATATGACAC TGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATTTCAAGGAA TACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCTCTGCAAGA TTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTACCATCCTG GAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGAAGACACC TACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCCACCCAAG GGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCTCAAAGAG AAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCTCTTGCAAG CAGGACTGCAAGCTAGACCTAAACTGTAA SEQIDNO.34:52L1N5nt ATGCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTTCTAAAGTGGTCT CCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCTGGTAGTTCAAG ACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGTCCTCAGGAAAC GGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACCGCGTCTTCCGTA TCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAGTTTCTATAACCC AGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAATTGGCAGGGGTC AACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAAGTTCGACGACA CAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAACCGTGAATGCC TCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGCAAGCCGCCTAT CGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCAGGAAACCCAG GAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGATGGTGACATGGT CGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTTCCAAGAGCGAT GTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTATCTGCAAATGGC TTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTC GTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCTGTCCCCGGAGAC CTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTGCAGTCTTCCGCTT TCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGCCAACTCTTTAATA AGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGCATCTGCTGGGGTA ACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCAATATGACACTGTG CGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATTTCAAGGAATACTT GCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCTCTGCAAGATTACT CTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTACCATCCTGGAGGA TTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGAAGACACCTACCG CTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCCACCCAAGGGTAA GGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTT CAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGG ACTGCAAGCTAGACCTAAACTGTAA SEQIDNO.35:52L1N8nt ATGGCTACTGTGTACTTGCCTCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATG AATACGTCTCACGTACCTCGATTTACTATTACGCTGGTAGTTCAAGACTGTTGAC AGTCGGCCACCCATACTTTTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAA GGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTG CCTGACCCCAACAAATTCGGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCC AGAGACTGGTGTGGGCCTGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTG GGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAAGTTCGACGACACAGAGACT TCTAACAAATACGCTGGTAAGCCAGGCATCGACAACCGTGAATGCCTCTCCATG GATTACAAACAGACCCAACTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAG CATTGGGGTAAAGGCACACCTTGCAACAATAACTCAGGAAACCCAGGAGACTG CCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGATGGTGACATGGTCGACACT GGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCA TCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTATCTGCAAATGGCTTCAGA ACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGT CACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATA TTCAAGGTTCCAACAGCGGTAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCC CAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGCCAACTCTTTAATAAGCCTT ACTGGTTGCAGAGGGCTCAAGGACACAACAATGGCATCTGCTGGGGTAACCAG CTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCAATATGACACTGTGCGCCG AGGTGAAGAAGGAATCCACATACAAAAACGAGAATTTCAAGGAATACTTGCGTC ACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCTCTGCAAGATTACTCTCAC CGCTGATGTTATGACATATATCCATAAGATGGACGCTACCATCCTGGAGGATTGGC AATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGAAGACACCTACCGCTTCG TCACAAGTACTGCCATTACTTGTCAGAAGAACACTCCACCCAAGGGTAAGGAGG ACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCG CCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGC AAGCTAGACCTAAACTGTAA SEQIDNO.36:52L1N10nt ATGGTGTACTTGCCTCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATGAATACG TCTCACGTACCTCGATTTACTATTACGCTGGTAGTTCAAGACTGTTGACAGTCGG CCACCCATACTTTTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAAGGTCCTT GTGCCGAAAGTTTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTGCCTGACC CCAACAAATTCGGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCCAGAGACT GGTGTGGGCCTGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTGGGCGTGG GAATCAGCGGTCACCCCCTTCTCAATAAGTTCGACGACACAGAGACTTCTAACA AATACGCTGGTAAGCCAGGCATCGACAACCGTGAATGCCTCTCCATGGATTACA AACAGACCCAACTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAGCATTGGG GTAAAGGCACACCTTGCAACAATAACTCAGGAAACCCAGGAGACTGCCCACCT TTGCAGCTTATCAACTCGGTTATTCAAGATGGTGACATGGTCGACACTGGCTTTG GATGTATGGACTTCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCATCGACAT CTGCTCTTCCGTGTGTAAATACCCAGATTATCTGCAAATGGCTTCAGAACCTTAC GGAGACTCTCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGTCACTTTT TCAACAGAGCCGGTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATATTCAAGG TTCCAACAGCGGTAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCCCAACTCC TTCAGGCAGCATGGTGACCAGTGAAAGCCAACTCTTTAATAAGCCTTACTGGTT GCAGAGGGCTCAAGGACACAACAATGGCATCTGCTGGGGTAACCAGCTGTTCG TTACAGTCGTCGATACCACTCGTTCTACCAATATGACACTGTGCGCCGAGGTGAA GAAGGAATCCACATACAAAAACGAGAATTTCAAGGAATACTTGCGTCACGGCGA GGAATTTGACCTTCAATTCATCTTCCAGCTCTGCAAGATTACTCTCACCGCTGAT GTTATGACATATATCCATAAGATGGACGCTACCATCCTGGAGGATTGGCAATTTGG ACTGACTCCCCCACCCTCAGCTTCGTTGGAAGACACCTACCGCTTCGTCACAAG TACTGCCATTACTTGTCAGAAGAACACTCCACCCAAGGGTAAGGAGGACCCACT TAAGGAGTACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCGCCGACCT GGATCAATTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGCAAGCTAG ACCTAAACTGTAA SEQIDNO.37:52L1N13nt ATGCCTCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATGAATACGTCTCACGTA CCTCGATTTACTATTACGCTGGTAGTTCAAGACTGTTGACAGTCGGCCACCCATA CTTTTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAA AGTTTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAA TTCGGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCCAGAGACTGGTGTGG GCCTGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTGGGCGTGGGAATCAG CGGTCACCCCCTTCTCAATAAGTTCGACGACACAGAGACTTCTAACAAATACGC TGGTAAGCCAGGCATCGACAACCGTGAATGCCTCTCCATGGATTACAAACAGAC CCAACTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAGCATTGGGGTAAAGG CACACCTTGCAACAATAACTCAGGAAACCCAGGAGACTGCCCACCTTTGCAGCT TATCAACTCGGTTATTCAAGATGGTGACATGGTCGACACTGGCTTTGGATGTATG GACTTCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCATCGACATCTGCTCTT CCGTGTGTAAATACCCAGATTATCTGCAAATGGCTTCAGAACCTTACGGAGACTC TCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGTCACTTTTTCAACAGA GCCGGTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATATTCAAGGTTCCAACA GCGGTAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCA GCATGGTGACCAGTGAAAGCCAACTCTTTAATAAGCCTTACTGGTTGCAGAGGG CTCAAGGACACAACAATGGCATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCG TCGATACCACTCGTTCTACCAATATGACACTGTGCGCCGAGGTGAAGAAGGAAT CCACATACAAAAACGAGAATTTCAAGGAATACTTGCGTCACGGCGAGGAATTTG ACCTTCAATTCATCTTCCAGCTCTGCAAGATTACTCTCACCGCTGATGTTATGACA TATATCCATAAGATGGACGCTACCATCCTGGAGGATTGGCAATTTGGACTGACTC CCCCACCCTCAGCTTCGTTGGAAGACACCTACCGCTTCGTCACAAGTACTGCCA TTACTTGTCAGAAGAACACTCCACCCAAGGGTAAGGAGGACCCACTTAAGGAG TACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCGCCGACCTGGATCAA TTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGCAAGCTAGACCTAAA CTGTAA SEQIDNO.38:52L1N15nt ATGGTACCTGTTTCTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGA TTTACTATTACGCTGGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCT ATCAAGAATACGTCCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCG GGTCTCCAATACCGCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCT TCCCAGATACTAGTTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCA CAGGACTCGAAATTGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCAC CCCCTTCTCAATAAGTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAG CCAGGCATCGACAACCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTG TGTATTCTGGGATGCAAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTT GCAACAATAACTCAGGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACT CGGTTATTCAAGATGGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAA TACTCTCCAGGCTTCCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGT AAATACCCAGATTATCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCT TCTTCTTGCGCAGGGAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTA CCTTGGGCGATCCTGTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAA CACAGCCACCGTGCAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGT GACCAGTGAAAGCCAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGG ACACAACAATGGCATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATAC CACTCGTTCTACCAATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATA CAAAAACGAGAATTTCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCA ATTCATCTTCCAGCTCTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCC ATAAGATGGACGCTACCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACC CTCAGCTTCGTTGGAAGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGT CAGAAGAACACTCCACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTT TTGGGAAGTGGATCTCAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCT GGGTCGTAAGTTCCTCTTGCAAGCAGGACTGCAAGCTAGACCTAAACTGTAA SEQIDNO.39:52L1N18nt ATGTCTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTA CGCTGGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAAT ACGTCCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAA TACCGCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATA CTAGTTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCG AAATTGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCA ATAAGTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCG ACAACCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGG ATGCAAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAA CTCAGGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCA AGATGGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAG GCTTCCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAG ATTATCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCG CAGGGAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGA TCCTGTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCAC CGTGCAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGA AAGCCAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAA TGGCATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCT ACCAATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGA GAATTTCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTC CAGCTCTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGA CGCTACCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCG TTGGAAGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAAC ACTCCACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTG GATCTCAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAG TTCCTCTTGCAAGCAGGACTGCAAGCTAGACCTAAACTGTAA SEQIDNO.40:52L1N20nt ATGGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCTGG TAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGTCC TCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACCGC GTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAGTT TCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAATTG GCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAAGT TCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAACC GTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGCAA GCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCAGG AAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGATGGT GACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTTCCA AGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTATCT GCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGGGA GCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCTGTC CCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTGCAG TCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGCCAA CTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGCATC TGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCAATA TGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATTTC AAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCTCT GCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTACC ATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGAA GACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCCA CCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCTC AAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCTC TTGCAAGCAGGACTGCAAGCTAGACCTAAACTGTAA SEQIDNO.41:52L1CS1nt ATGTCCGTGTGGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTT CTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCT GGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGT CCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACC GCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAG TTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAAT TGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAA GTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAA CCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGC AAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCA GGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGAT GGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTT CCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTA TCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGG GAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCT GTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTG CAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGC CAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGC ATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCA ATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATT TCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCT CTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTA CCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGA AGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCC ACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCT CAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCT CTTGCAAGCAGGACTGCAAGCTCGTCCTGGACTGAAAGGTCCTGCATCGAGCG CTCCTAGAACGTCGACGGACGGCTCGGGAGTGGGACGCTAA SEQIDNO.42:52L1CS2nt ATGTCCGTGTGGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTT CTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCT GGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGT CCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACC GCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAG TTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAAT TGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAA GTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAA CCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGC AAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCA GGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGAT GGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTT CCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTA TCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGG GAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCT GTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTG CAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGC CAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGC ATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCA ATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATT TCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCT CTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTA CCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGA AGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCC ACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCT CAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCT CTTGCAAGCAGGACTGCAAGCTCGTCCTGGACTGAAAGGTCCTGCATCGAGCG CTCCTAGAACGTCGACGGACGGCTCGGGAGTGGACGGCTAA SEQIDNO.43:52L1CS3nt ATGTCCGTGTGGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTT CTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCT GGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGT CCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACC GCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAG TTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAAT TGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAA GTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAA CCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGC AAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCA GGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGAT GGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTT CCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTA TCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGG GAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCT GTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTG CAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGC CAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGC ATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCA ATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATT TCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCT CTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTA CCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGA AGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCC ACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCT CAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCT CTTGCAAGCAGGACTGCAAGCTCGTCCTGGACTGGGATCGCCTGCATCGAGCGC TCCTAGAACGTCGACGGACGGCTCGGGAGTGAAACGCTAA SEQIDNO.44:52L1CS4nt ATGTCCGTGTGGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTT CTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCT GGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGT CCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACC GCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAG TTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAAT TGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAA GTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAA CCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGC AAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCA GGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGAT GGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTT CCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTA TCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGG GAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCT GTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTG CAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGC CAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGC ATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCA ATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATT TCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCT CTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTA CCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGA AGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCC ACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCT CAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCT CTTGCAAGCAGGACTGCAAGCTCGTCCTGGACTGGGATCGCCTGCATCGAGCGC TCCTAGAACGTCGACGGACGGCTCGGGAGTGGACCGCTAA SEQIDNO.45:52L1CS5nt ATGTCCGTGTGGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTT CTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCT GGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGT CCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACC GCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAG TTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAAT TGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAA GTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAA CCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGC AAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCA GGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGAT GGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTT CCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTA TCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGG GAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCT GTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTG CAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGC CAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGC ATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCA ATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATT TCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCT CTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTA CCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGA AGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCC ACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCT CAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCT CTTGCAAGCAGGACTGCAAGCTAGACCTAAACTGGCTGGTCCTGCCTCTTCCGC ACCCGCGACTTCAACCGCTGCCGGCGGAGTTGGGTCGTAA SEQIDNO.46:52L1CS6nt ATGTCCGTGTGGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTT CTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCT GGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGT CCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACC GCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAG TTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAAT TGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAA GTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAA CCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGC AAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCA GGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGAT GGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTT CCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTA TCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGG GAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCT GTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTG CAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGC CAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGC ATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCA ATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATT TCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCT CTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTA CCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGA AGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCC ACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCT CAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCT CTTGCAAGCAGGACTGCAAGCTAGACCTAAACTGGAAGCTCCTGCCTCTTCCGC ACCCGGTACTTCAACCGGCTCGAAAGCGGTTGCTGGATAA SEQIDNO.47:52L1CS7nt ATGTCCGTGTGGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTT CTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCT GGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGT CCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACC GCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAG TTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAAT TGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAA GTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAA CCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGC AAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCA GGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGAT GGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTT CCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTA TCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGG GAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCT GTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTG CAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGC CAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGC ATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCA ATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATT TCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCT CTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTA CCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGA AGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCC ACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCT CAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCT CTTGCAAGCAGGACTGCAAGCTAGACCTAAACTGGCTGGTCCTGCTTCCTCAGC TCCAGCTACCTCAACCGACGGTTCTGGTGTGAAGCGCTAA SEQIDNO.48:52L1CS8nt ATGTCCGTGTGGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTT CTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCT GGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGT CCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACC GCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAG TTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAAT TGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAA GTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAA CCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGC AAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCA GGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGAT GGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTT CCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTA TCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGG GAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCT GTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTG CAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGC CAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGC ATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCA ATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATT TCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCT CTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTA CCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGA AGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCC ACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCT CAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCT CTTGCAAGCAGGACTGCAAGCTAGACCTAAACTGGCTGGTCCTGCTTCCTCAGC TCCACGTACCTCAACCGACGGTTCTGGTGTGAAGCGCTAA SEQIDNO.49:52L1CS9nt ATGTCCGTGTGGCGTCCTTCCGAGGCTACTGTGTACTTGCCTCCAGTACCTGTTT CTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCTCGATTTACTATTACGCT GGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTTTTCTATCAAGAATACGT CCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACC GCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTCGGCTTCCCAGATACTAG TTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCCTGCACAGGACTCGAAAT TGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAA GTTCGACGACACAGAGACTTCTAACAAATACGCTGGTAAGCCAGGCATCGACAA CCGTGAATGCCTCTCCATGGATTACAAACAGACCCAACTGTGTATTCTGGGATGC AAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACACCTTGCAACAATAACTCA GGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGAT GGTGACATGGTCGACACTGGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTT CCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTA TCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGG GAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCT GTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGGTAACACAGCCACCGTG CAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGC CAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAAGGACACAACAATGGC ATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCA ATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACATACAAAAACGAGAATT TCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCT CTGCAAGATTACTCTCACCGCTGATGTTATGACATATATCCATAAGATGGACGCTA CCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGA AGACACCTACCGCTTCGTCACAAGTACTGCCATTACTTGTCAGAAGAACACTCC ACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCT CAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCT CTTGCAAGCAGGACTGCAAGCGGGTCCTGGCTTGTCGGGTCCTGCCTCGAGCGC CCCTAGAACGTCGACGGGTGGCTCGGCCGTGGGTAGCTAA SEQIDNO.50:52L1N13CS1nt ATGCCTCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATGAATACGTCTCACGTA CCTCGATTTACTATTACGCTGGTAGTTCAAGACTGTTGACAGTCGGCCACCCATA CTTTTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAA AGTTTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAA TTCGGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCCAGAGACTGGTGTGG GCCTGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTGGGCGTGGGAATCAG CGGTCACCCCCTTCTCAATAAGTTCGACGACACAGAGACTTCTAACAAATACGC TGGTAAGCCAGGCATCGACAACCGTGAATGCCTCTCCATGGATTACAAACAGAC CCAACTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAGCATTGGGGTAAAGG CACACCTTGCAACAATAACTCAGGAAACCCAGGAGACTGCCCACCTTTGCAGCT TATCAACTCGGTTATTCAAGATGGTGACATGGTCGACACTGGCTTTGGATGTATG GACTTCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCATCGACATCTGCTCTT CCGTGTGTAAATACCCAGATTATCTGCAAATGGCTTCAGAACCTTACGGAGACTC TCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGTCACTTTTTCAACAGA GCCGGTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATATTCAAGGTTCCAACA GCGGTAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCA GCATGGTGACCAGTGAAAGCCAACTCTTTAATAAGCCTTACTGGTTGCAGAGGG CTCAAGGACACAACAATGGCATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCG TCGATACCACTCGTTCTACCAATATGACACTGTGCGCCGAGGTGAAGAAGGAAT CCACATACAAAAACGAGAATTTCAAGGAATACTTGCGTCACGGCGAGGAATTTG ACCTTCAATTCATCTTCCAGCTCTGCAAGATTACTCTCACCGCTGATGTTATGACA TATATCCATAAGATGGACGCTACCATCCTGGAGGATTGGCAATTTGGACTGACTC CCCCACCCTCAGCTTCGTTGGAAGACACCTACCGCTTCGTCACAAGTACTGCCA TTACTTGTCAGAAGAACACTCCACCCAAGGGTAAGGAGGACCCACTTAAGGAG TACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCGCCGACCTGGATCAA TTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGCAAGCGAGACCTGGC TTGTCGGGTCCTGCCTCGAGCGCCCCTAGAACGTCGACGGGTGGCTCGGCCGTG GGTAGCTAA SEQIDNO.51:52L1N13CS2nt ATGCCTCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATGAATACGTCTCACGTA CCTCGATTTACTATTACGCTGGTAGTTCAAGACTGTTGACAGTCGGCCACCCATA CTTTTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAA AGTTTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAA TTCGGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCCAGAGACTGGTGTGG GCCTGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTGGGCGTGGGAATCAG CGGTCACCCCCTTCTCAATAAGTTCGACGACACAGAGACTTCTAACAAATACGC TGGTAAGCCAGGCATCGACAACCGTGAATGCCTCTCCATGGATTACAAACAGAC CCAACTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAGCATTGGGGTAAAGG CACACCTTGCAACAATAACTCAGGAAACCCAGGAGACTGCCCACCTTTGCAGCT TATCAACTCGGTTATTCAAGATGGTGACATGGTCGACACTGGCTTTGGATGTATG GACTTCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCATCGACATCTGCTCTT CCGTGTGTAAATACCCAGATTATCTGCAAATGGCTTCAGAACCTTACGGAGACTC TCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGTCACTTTTTCAACAGA GCCGGTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATATTCAAGGTTCCAACA GCGGTAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCA GCATGGTGACCAGTGAAAGCCAACTCTTTAATAAGCCTTACTGGTTGCAGAGGG CTCAAGGACACAACAATGGCATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCG TCGATACCACTCGTTCTACCAATATGACACTGTGCGCCGAGGTGAAGAAGGAAT CCACATACAAAAACGAGAATTTCAAGGAATACTTGCGTCACGGCGAGGAATTTG ACCTTCAATTCATCTTCCAGCTCTGCAAGATTACTCTCACCGCTGATGTTATGACA TATATCCATAAGATGGACGCTACCATCCTGGAGGATTGGCAATTTGGACTGACTC CCCCACCCTCAGCTTCGTTGGAAGACACCTACCGCTTCGTCACAAGTACTGCCA TTACTTGTCAGAAGAACACTCCACCCAAGGGTAAGGAGGACCCACTTAAGGAG TACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCGCCGACCTGGATCAA TTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGCAAGCGGGTCCTGGC TTGTCGGGTCCTGCCTCGAGCGCCCCTAGAACGTCGACGGGTGGCTCGGCCGTG GGTAGCTAA SEQIDNO.52:52L1N13CS3nt ATGCCTCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATGAATACGTCTCACGTA CCTCGATTTACTATTACGCTGGTAGTTCAAGACTGTTGACAGTCGGCCACCCATA CTTTTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAA AGTTTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAA TTCGGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCCAGAGACTGGTGTGG GCCTGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTGGGCGTGGGAATCAG CGGTCACCCCCTTCTCAATAAGTTCGACGACACAGAGACTTCTAACAAATACGC TGGTAAGCCAGGCATCGACAACCGTGAATGCCTCTCCATGGATTACAAACAGAC CCAACTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAGCATTGGGGTAAAGG CACACCTTGCAACAATAACTCAGGAAACCCAGGAGACTGCCCACCTTTGCAGCT TATCAACTCGGTTATTCAAGATGGTGACATGGTCGACACTGGCTTTGGATGTATG GACTTCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCATCGACATCTGCTCTT CCGTGTGTAAATACCCAGATTATCTGCAAATGGCTTCAGAACCTTACGGAGACTC TCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGTCACTTTTTCAACAGA GCCGGTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATATTCAAGGTTCCAACA GCGGTAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCA GCATGGTGACCAGTGAAAGCCAACTCTTTAATAAGCCTTACTGGTTGCAGAGGG CTCAAGGACACAACAATGGCATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCG TCGATACCACTCGTTCTACCAATATGACACTGTGCGCCGAGGTGAAGAAGGAAT CCACATACAAAAACGAGAATTTCAAGGAATACTTGCGTCACGGCGAGGAATTTG ACCTTCAATTCATCTTCCAGCTCTGCAAGATTACTCTCACCGCTGATGTTATGACA TATATCCATAAGATGGACGCTACCATCCTGGAGGATTGGCAATTTGGACTGACTC CCCCACCCTCAGCTTCGTTGGAAGACACCTACCGCTTCGTCACAAGTACTGCCA TTACTTGTCAGAAGAACACTCCACCCAAGGGTAAGGAGGACCCACTTAAGGAG TACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCGCCGACCTGGATCAA TTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGCAAGCTAGACCTAAA CTGGCCGGTCCTGCCTCGAGCGCCCCTGCCACGTCGACGGCTGCGGGAGGCGT GGGTAGCTAA SEQIDNO.53:52L1NS1C19nt ATGCCTAGCGAGGCTACCCCTCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATG AATACGTCTCACGTACCTCGATTTACTATTACGCTGGTAGTTCAAGACTGTTGAC AGTCGGCCACCCATACTTTTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAA GGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTG CCTGACCCCAACAAATTCGGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCC AGAGACTGGTGTGGGCCTGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTG GGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAAGTTCGACGACACAGAGACT TCTAACAAATACGCTGGTAAGCCAGGCATCGACAACCGTGAATGCCTCTCCATG GATTACAAACAGACCCAACTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAG CATTGGGGTAAAGGCACACCTTGCAACAATAACTCAGGAAACCCAGGAGACTG CCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGATGGTGACATGGTCGACACT GGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCA TCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTATCTGCAAATGGCTTCAGA ACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGT CACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATA TTCAAGGTTCCAACAGCGGTAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCC CAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGCCAACTCTTTAATAAGCCTT ACTGGTTGCAGAGGGCTCAAGGACACAACAATGGCATCTGCTGGGGTAACCAG CTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCAATATGACACTGTGCGCCG AGGTGAAGAAGGAATCCACATACAAAAACGAGAATTTCAAGGAATACTTGCGTC ACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCTCTGCAAGATTACTCTCAC CGCTGATGTTATGACATATATCCATAAGATGGACGCTACCATCCTGGAGGATTGGC AATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGAAGACACCTACCGCTTCG TCACAAGTACTGCCATTACTTGTCAGAAGAACACTCCACCCAAGGGTAAGGAGG ACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCG CCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGC AAGCTAGACCTAAACTGTAA SEQIDNO.54:52L1INS1C25 ATGCCTAGCGAGGCTACCCCTCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATG AATACGTCTCACGTACCTCGATTTACTATTACGCTGGTAGTTCAAGACTGTTGAC AGTCGGCCACCCATACTTTTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAA GGTCCTTGTGCCGAAAGTTTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTG CCTGACCCCAACAAATTCGGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCC AGAGACTGGTGTGGGCCTGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTG GGCGTGGGAATCAGCGGTCACCCCCTTCTCAATAAGTTCGACGACACAGAGACT TCTAACAAATACGCTGGTAAGCCAGGCATCGACAACCGTGAATGCCTCTCCATG GATTACAAACAGACCCAACTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAG CATTGGGGTAAAGGCACACCTTGCAACAATAACTCAGGAAACCCAGGAGACTG CCCACCTTTGCAGCTTATCAACTCGGTTATTCAAGATGGTGACATGGTCGACACT GGCTTTGGATGTATGGACTTCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCA TCGACATCTGCTCTTCCGTGTGTAAATACCCAGATTATCTGCAAATGGCTTCAGA ACCTTACGGAGACTCTCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGT CACTTTTTCAACAGAGCCGGTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATA TTCAAGGTTCCAACAGCGGTAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCC CAACTCCTTCAGGCAGCATGGTGACCAGTGAAAGCCAACTCTTTAATAAGCCTT ACTGGTTGCAGAGGGCTCAAGGACACAACAATGGCATCTGCTGGGGTAACCAG CTGTTCGTTACAGTCGTCGATACCACTCGTTCTACCAATATGACACTGTGCGCCG AGGTGAAGAAGGAATCCACATACAAAAACGAGAATTTCAAGGAATACTTGCGTC ACGGCGAGGAATTTGACCTTCAATTCATCTTCCAGCTCTGCAAGATTACTCTCAC CGCTGATGTTATGACATATATCCATAAGATGGACGCTACCATCCTGGAGGATTGGC AATTTGGACTGACTCCCCCACCCTCAGCTTCGTTGGAAGACACCTACCGCTTCG TCACAAGTACTGCCATTACTTGTCAGAAGAACACTCCACCCAAGGGTAAGGAGG ACCCACTTAAGGAGTACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCG CCGACCTGGATCAATTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGTA A SEQIDNO.55:52L1NS2C19nt ATGTCCGAGCGTCCTCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATGAATACG TCTCACGTACCTCGATTTACTATTACGCTGGTAGTTCAAGACTGTTGACAGTCGG CCACCCATACTTTTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAAGGTCCTT GTGCCGAAAGTTTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTGCCTGACC CCAACAAATTCGGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCCAGAGACT GGTGTGGGCCTGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTGGGCGTGG GAATCAGCGGTCACCCCCTTCTCAATAAGTTCGACGACACAGAGACTTCTAACA AATACGCTGGTAAGCCAGGCATCGACAACCGTGAATGCCTCTCCATGGATTACA AACAGACCCAACTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAGCATTGGG GTAAAGGCACACCTTGCAACAATAACTCAGGAAACCCAGGAGACTGCCCACCT TTGCAGCTTATCAACTCGGTTATTCAAGATGGTGACATGGTCGACACTGGCTTTG GATGTATGGACTTCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCATCGACAT CTGCTCTTCCGTGTGTAAATACCCAGATTATCTGCAAATGGCTTCAGAACCTTAC GGAGACTCTCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGTCACTTTT TCAACAGAGCCGGTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATATTCAAGG TTCCAACAGCGGTAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCCCAACTCC TTCAGGCAGCATGGTGACCAGTGAAAGCCAACTCTTTAATAAGCCTTACTGGTT GCAGAGGGCTCAAGGACACAACAATGGCATCTGCTGGGGTAACCAGCTGTTCG TTACAGTCGTCGATACCACTCGTTCTACCAATATGACACTGTGCGCCGAGGTGAA GAAGGAATCCACATACAAAAACGAGAATTTCAAGGAATACTTGCGTCACGGCGA GGAATTTGACCTTCAATTCATCTTCCAGCTCTGCAAGATTACTCTCACCGCTGAT GTTATGACATATATCCATAAGATGGACGCTACCATCCTGGAGGATTGGCAATTTGG ACTGACTCCCCCACCCTCAGCTTCGTTGGAAGACACCTACCGCTTCGTCACAAG TACTGCCATTACTTGTCAGAAGAACACTCCACCCAAGGGTAAGGAGGACCCACT TAAGGAGTACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCGCCGACCT GGATCAATTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGCAAGCTAG ACCTAAACTGTAA SEQIDNO.56:52L1NS3C19nt ATGTCCGAGCCTCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATGAATACGTCT CACGTACCTCGATTTACTATTACGCTGGTAGTTCAAGACTGTTGACAGTCGGCCA CCCATACTTTTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAAGGTCCTTGTG CCGAAAGTTTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTGCCTGACCCCA ACAAATTCGGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCCAGAGACTGGT GTGGGCCTGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTGGGCGTGGGAA TCAGCGGTCACCCCCTTCTCAATAAGTTCGACGACACAGAGACTTCTAACAAAT ACGCTGGTAAGCCAGGCATCGACAACCGTGAATGCCTCTCCATGGATTACAAAC AGACCCAACTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAGCATTGGGGTA AAGGCACACCTTGCAACAATAACTCAGGAAACCCAGGAGACTGCCCACCTTTG CAGCTTATCAACTCGGTTATTCAAGATGGTGACATGGTCGACACTGGCTTTGGAT GTATGGACTTCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCATCGACATCTG CTCTTCCGTGTGTAAATACCCAGATTATCTGCAAATGGCTTCAGAACCTTACGGA GACTCTCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGTCACTTTTTCA ACAGAGCCGGTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATATTCAAGGTTC CAACAGCGGTAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCCCAACTCCTTC AGGCAGCATGGTGACCAGTGAAAGCCAACTCTTTAATAAGCCTTACTGGTTGCA GAGGGCTCAAGGACACAACAATGGCATCTGCTGGGGTAACCAGCTGTTCGTTAC AGTCGTCGATACCACTCGTTCTACCAATATGACACTGTGCGCCGAGGTGAAGAA GGAATCCACATACAAAAACGAGAATTTCAAGGAATACTTGCGTCACGGCGAGGA ATTTGACCTTCAATTCATCTTCCAGCTCTGCAAGATTACTCTCACCGCTGATGTTA TGACATATATCCATAAGATGGACGCTACCATCCTGGAGGATTGGCAATTTGGACT GACTCCCCCACCCTCAGCTTCGTTGGAAGACACCTACCGCTTCGTCACAAGTAC TGCCATTACTTGTCAGAAGAACACTCCACCCAAGGGTAAGGAGGACCCACTTAA GGAGTACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCGCCGACCTGGA TCAATTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGCAAGCTAGACCT AAACTGTAA SEQIDNO.57:52L1NS4C19nt ATGTCCCCTCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATGAATACGTCTCAC GTACCTCGATTTACTATTACGCTGGTAGTTCAAGACTGTTGACAGTCGGCCACCC ATACTTTTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAAGGTCCTTGTGCCG AAAGTTTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTGCCTGACCCCAAC AAATTCGGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCCAGAGACTGGTGT GGGCCTGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTGGGCGTGGGAATC AGCGGTCACCCCCTTCTCAATAAGTTCGACGACACAGAGACTTCTAACAAATAC GCTGGTAAGCCAGGCATCGACAACCGTGAATGCCTCTCCATGGATTACAAACAG ACCCAACTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAGCATTGGGGTAAA GGCACACCTTGCAACAATAACTCAGGAAACCCAGGAGACTGCCCACCTTTGCA GCTTATCAACTCGGTTATTCAAGATGGTGACATGGTCGACACTGGCTTTGGATGT ATGGACTTCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCATCGACATCTGCT CTTCCGTGTGTAAATACCCAGATTATCTGCAAATGGCTTCAGAACCTTACGGAGA CTCTCTGTTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGTCACTTTTTCAAC AGAGCCGGTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATATTCAAGGTTCCA ACAGCGGTAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCCCAACTCCTTCAG GCAGCATGGTGACCAGTGAAAGCCAACTCTTTAATAAGCCTTACTGGTTGCAGA GGGCTCAAGGACACAACAATGGCATCTGCTGGGGTAACCAGCTGTTCGTTACAG TCGTCGATACCACTCGTTCTACCAATATGACACTGTGCGCCGAGGTGAAGAAGG AATCCACATACAAAAACGAGAATTTCAAGGAATACTTGCGTCACGGCGAGGAAT TTGACCTTCAATTCATCTTCCAGCTCTGCAAGATTACTCTCACCGCTGATGTTATG ACATATATCCATAAGATGGACGCTACCATCCTGGAGGATTGGCAATTTGGACTGA CTCCCCCACCCTCAGCTTCGTTGGAAGACACCTACCGCTTCGTCACAAGTACTG CCATTACTTGTCAGAAGAACACTCCACCCAAGGGTAAGGAGGACCCACTTAAGG AGTACATGTTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCGCCGACCTGGATC AATTTCCTCTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGCAAGCTAGACCTA AACTGTAA SEQIDNO.58:52L1N14C25nt ATGCCAGTACCTGTTTCTAAAGTGGTCTCCACTGATGAATACGTCTCACGTACCT CGATTTACTATTACGCTGGTAGTTCAAGACTGTTGACAGTCGGCCACCCATACTT TTCTATCAAGAATACGTCCTCAGGAAACGGTAAGAAGGTCCTTGTGCCGAAAGT TTCGGGTCTCCAATACCGCGTCTTCCGTATCAAGCTGCCTGACCCCAACAAATTC GGCTTCCCAGATACTAGTTTCTATAACCCAGAGACCCAGAGACTGGTGTGGGCC TGCACAGGACTCGAAATTGGCAGGGGTCAACCTTTGGGCGTGGGAATCAGCGG TCACCCCCTTCTCAATAAGTTCGACGACACAGAGACTTCTAACAAATACGCTGGT AAGCCAGGCATCGACAACCGTGAATGCCTCTCCATGGATTACAAACAGACCCAA CTGTGTATTCTGGGATGCAAGCCGCCTATCGGTGAGCATTGGGGTAAAGGCACA CCTTGCAACAATAACTCAGGAAACCCAGGAGACTGCCCACCTTTGCAGCTTATC AACTCGGTTATTCAAGATGGTGACATGGTCGACACTGGCTTTGGATGTATGGACT TCAATACTCTCCAGGCTTCCAAGAGCGATGTCCCCATCGACATCTGCTCTTCCGT GTGTAAATACCCAGATTATCTGCAAATGGCTTCAGAACCTTACGGAGACTCTCTG TTCTTCTTCTTGCGCAGGGAGCAGATGTTCGTTCGTCACTTTTTCAACAGAGCCG GTACCTTGGGCGATCCTGTCCCCGGAGACCTTTATATTCAAGGTTCCAACAGCGG TAACACAGCCACCGTGCAGTCTTCCGCTTTCTTCCCAACTCCTTCAGGCAGCATG GTGACCAGTGAAAGCCAACTCTTTAATAAGCCTTACTGGTTGCAGAGGGCTCAA GGACACAACAATGGCATCTGCTGGGGTAACCAGCTGTTCGTTACAGTCGTCGAT ACCACTCGTTCTACCAATATGACACTGTGCGCCGAGGTGAAGAAGGAATCCACA TACAAAAACGAGAATTTCAAGGAATACTTGCGTCACGGCGAGGAATTTGACCTT CAATTCATCTTCCAGCTCTGCAAGATTACTCTCACCGCTGATGTTATGACATATAT CCATAAGATGGACGCTACCATCCTGGAGGATTGGCAATTTGGACTGACTCCCCCA CCCTCAGCTTCGTTGGAAGACACCTACCGCTTCGTCACAAGTACTGCCATTACTT GTCAGAAGAACACTCCACCCAAGGGTAAGGAGGACCCACTTAAGGAGTACATG TTTTGGGAAGTGGATCTCAAAGAGAAGTTCAGCGCCGACCTGGATCAATTTCCT CTGGGTCGTAAGTTCCTCTTGCAAGCAGGACTGTAA.