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Recombinant foot-and-mouth disease virus VP1 confluent protein vaccine

A foot-and-mouth disease virus and fusion protein technology, which is applied in the field of genetic engineering recombinant protein vaccines and recombinant foot-and-mouth disease virus VP1 fusion protein vaccines, can solve problems such as insufficient inactivation

Inactive Publication Date: 2006-12-20
BEIJING HYDVAX BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional foot-and-mouth disease vaccine is an inactivated vaccine made by emulsifying the initially purified inactivated foot-and-mouth disease virus with an adjuvant. It has a considerable protective effect on susceptible livestock, but it may cause foot-and-mouth disease due to insufficient inactivation

Method used

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  • Recombinant foot-and-mouth disease virus VP1 confluent protein vaccine
  • Recombinant foot-and-mouth disease virus VP1 confluent protein vaccine
  • Recombinant foot-and-mouth disease virus VP1 confluent protein vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Embodiment 1, synthetic VP1BT1 fusion peptide coding gene

[0069] The gene encoding VP1 BT1 fusion peptide was synthesized by PCR method. PCR primers with the following sequences were designed and synthesized:

[0070] Primer 1: 'CTGCAAGTACTGGCTCAAAAAGCAGAACGTGCTCTGCCGGGTGGCGAAGAAAACTACGGTGGT3'

[0071] Primer 2: 5′TGAAGGAGATGTCAGTGTGCTGACGACGCTGAACCTGAGTTTCACCACCGTAGTTTTCTTCGCC3′

[0072] Primer 3: 5'GAATTCAGATCTGGCGGTGTATCTAACGTACGTGGTGACCTGCAAGTACTGGCTCAA3'

[0073] Primer 4: 5′AAGCTTCCCGGAGTAACTTTAACGAAACGGTCCAGGATGAAGGAGATGTCAGTGTG3′

[0074] Using primers 1 and 2 to synthesize a DNA fragment (fragment 1, 2) with the following sequence:

[0075] CTGCAAGTAC TGGCTCAAAA AGCAGAACGT GCTCTGCCGG GTGGCGAAGA AAACTACGGT GGTGAAACTC

[0076] AGGTTCAGCG TCGTCAGCAC ACTGACATCT CCTTCA

[0077] Using fragments 1 and 2 as templates, primers 3 and 4 were used to synthesize the VP1 BT1 fusion peptide coding gene with restriction endonuclease cutting sites on both sides, and its...

Embodiment 2

[0105] Embodiment 2, synthetic VP1BT2 fusion peptide coding gene

[0106] The gene encoding VP1 BT1 fusion peptide was synthesized by PCR method. PCR primers with the following sequences were designed and synthesized:

[0107] Primer 7: 5'GTGACCTGCAAGTACTGGCTCAAAAAGCTAAACGTGCTCTGCCGGGTGGTCCGTCT3'

[0108] Primer 8: 5′CGGAGCAACGATTTTCTGTTTGTGACGAGCGTCAGACGGACCACCCGGCA3′

[0109] Primer 9: 5'CCATGGAATTCAGATCTGGTGGTGTTTCTAACGTTCGTGGTGACCTGCAAGTACTGGC3'

[0110] Primer 10: 5′AAGCTTGGATCCCAGGTCGAAGTTCAGCAGCTGTTTAGCCGGAGCAACGATTTTCTG3′

[0111] Adopt primer 7,8 to synthesize the DNA fragment (fragment 7,8) that has following sequence:

[0112] GTGACCTGCA AGTACTGGCT CAAAAAGCTA AACGTGCTCT GCCGGGTGGT CCGTCTGACG CTCGTCACAA

[0113] ACAGAAAATC GTTGCTCCG

[0114] Using fragments 7 and 8 as templates, primers 9 and 10 were used to synthesize the VP1 BT2 fusion peptide coding gene with restriction endonuclease cutting points on both sides, which has the following sequence as shown in ...

Embodiment 3

[0119] Example 3, Construction of VP1 peptide fusion protein encoding gene

[0120] The VP1BT1 fusion peptide coding gene on the pMD18-T Vect plasmid was digested with BamHI, the BT2 fusion peptide coding gene on the pMD18-TVect plasmid was digested with Bg1II, and the two digested fragments were ligated with T4 ligase to obtain the BT1 fusion peptide coding gene-BT2 fusion Peptide coding gene fusion gene (SEQ ID NO: 5), its sequence is as follows:

[0121] GAATTCAGAT CTGGCGGTGT ATCTAACGTA CGTGGTGACC TGCAAGTACT GGCTCAAAAA GCAGAACGTG

[0122] CTCTGCCGGG TGGCGAAGAA AACTACGGTG GTGAAACTCA GGTTCAGCGT CGTCAGCACA CTGACATCTC

[0123] CTTCATCCTG GACCGTTTCG TTAAAGTTAC TCCGGGATCT GGTGGTGTTT CTAACGTTCG TGGTGACCTG

[0124] CAAGTACTGG CTCAAAAAGC TAAACGTGCT CTGCCGGGTG GTCCGTCTGA CGCTCGTCAC AAACAGAAAA

[0125] TCGTTGCTCC GGCTAAACAG CTGCTGAACT TCGACCTGGG ATCCAAGCTT

[0126] The VP1 BT1 fusion peptide encoding gene-VP1 BT2 fusion peptide encoding gene fusion gene was digested with BamHI and...

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Abstract

Said invention provides recomposed foot-and -mouth disease virus VP1 fusion protein vaccine, which fuses peptide section of foot-and -mouth disease virus VP1, 2-polyglycine and 6-polyhestidine coded gene and recomposes fusion protein in bacillus coli. Said invention also provides amino acid sequence of recomposed foot-and -mouth disease virus VP1 fusion protein and nucleotide sequence, and effect for preventing foot-and -mouth disease virus infection.

Description

field of invention [0001] The present invention relates to the field of genetic engineering, in particular to genetic engineering recombinant protein vaccines, in particular to a recombinant foot-and-mouth disease virus VP1 fusion protein vaccine (hereinafter sometimes referred to as recombinant foot-and-mouth disease virus VP1 fusion protein) for preventing animal foot-and-mouth disease virus infection. It is a recombinant fusion protein produced in Escherichia coli by fusing the VP1 peptide of foot-and-mouth disease virus, 2-polyglycine, and 6-polyhistidine encoding genes. After being applied to animal organisms, it can induce the body to produce neutralizing antibodies against foot-and-mouth disease virus. It has the biological activity of preventing animal foot-and-mouth disease virus infection. Background of the invention [0002] Foot-and-mouth disease (FMD) is a severe infectious disease caused by foot-and-mouth disease virus. Pigs, cattle, and sheep can all be infec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/135A61P31/14A61K39/00A61P31/12C07K14/09C12N15/42C12N15/62
CPCC07K14/005A61K39/00C07K2319/00C12N2770/32122A61P31/12A61P31/14
Inventor 于永利邵明玉王丽颖
Owner BEIJING HYDVAX BIOTECH
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