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Foot-and-mouth disease virus antigen, gene capable of expressing foot-and-mouth disease virus antigen, recombinant vector comprising gene, and recombinant lactococcus lactis

A technology for foot-and-mouth disease virus and Lactococcus lactis, which is applied in the directions of virus antigen components, viruses/phages, viruses, etc., can solve the problems of low efficiency of mucosal vaccines and achieve the effect of good immunogenicity

Inactive Publication Date: 2019-12-03
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The object of the present invention is to provide a kind of foot-and-mouth disease virus antigen, the gene TB1-Co1 expressing the foot-and-mouth disease virus antigen and its recombinant vector and recombinant Lactococcus lactis. mucosal vaccine

Method used

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  • Foot-and-mouth disease virus antigen, gene capable of expressing foot-and-mouth disease virus antigen, recombinant vector comprising gene, and recombinant lactococcus lactis
  • Foot-and-mouth disease virus antigen, gene capable of expressing foot-and-mouth disease virus antigen, recombinant vector comprising gene, and recombinant lactococcus lactis
  • Foot-and-mouth disease virus antigen, gene capable of expressing foot-and-mouth disease virus antigen, recombinant vector comprising gene, and recombinant lactococcus lactis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Obtaining of TB1 and TB1-Co1 genes

[0046] 1 Materials and methods

[0047] 1.1 Materials and sources

[0048] Lactococcus lactis NZ9000 and expression plasmid pNZ8148 were purchased from MoBiTec, Germany, and competent Escherichia coli Top10 and cloning vector pUC57 were purchased from Nanjing GenScript Biotechnology Co., Ltd.

[0049] 1.2 Test method

[0050] 1.2.1 Gene modification and synthesis

[0051] In eukaryotes or prokaryotes, proteins are either secreted outside the cell or stored inside the cell after translation and synthesis. Signal peptides are determinants of protein secretion out of the cell. The signal peptide is an amino acid sequence located at the N-terminus of the secretory protein, usually consisting of 21 to 30 amino acid residues. When synthesized, it is transcribed and translated together with the secretory peptide to form a propeptide. After the propeptide is correctly positioned, the signal peptide is captured by the signal pep...

Embodiment 2

[0061] Example 2 Obtaining of recombinant expression vectors pNZ8148-TB1 and pNZ8148-TB1-Co1

[0062] 1 Materials and methods

[0063] 1.1 Materials and sources

[0064] Restriction endonucleases Nco I and Hind III, and T4 DNA ligase were purchased from NEB Company; DNA Marker DL5,000 was purchased from Dalian Bao Biological Company; lysozyme was purchased from American Amresco Company; M17 dry powder medium, agar powder, and glucose were all purchased Purchased from OXOID Company in the United Kingdom; the plasmid mini-prep (Gram-negative bacteria) kit and DNA gel recovery kit were purchased from OmegaBio-Tek in the United States; the plasmid mini-prep (Gram-positive bacteria) kit was purchased from Beijing Suo Lai Bao Technology Co., Ltd.; the cloning vector pUC57 was provided by Nanjing GenScript Biotechnology Co., Ltd.

[0065] 1.2 Test method

[0066] Recombinant plasmids pUC57-TB1 and pUC57-TB1-Co1 of Example 1 were respectively digested with Nco I / Hind III small frag...

Embodiment 3

[0069] Embodiment 3 expresses the preparation and detection of type A foot-and-mouth disease virus antigen TB1 and TB1-Co1 recombinant bacteria

[0070] 1 Materials and methods

[0071] 1.1 Materials and sources

[0072] (1) Inducer: Nisin was purchased from Sigma, USA;

[0073] (2) Antibiotics: ampicillin (Amr) and chloramphenicol (Chr) were purchased from American Amresco Company.

[0074] 1.2 Test method

[0075] 1.2.1 Electrotransformation of the target gene in Lactococcus lactis NZ9000 and screening of resistant strains: the electrotransformed Lactococcus lactis NZ9000 was rejuvenated in a constant temperature water bath at 30°C for 3 hours, and then 1 mL of the bacterial solution was concentrated into 200 μL Spread on GM17 solid medium containing 10 μg / mL chloramphenicol (sealed), and culture in a constant temperature incubator at 30°C for 48h. The colonies on the plate were picked and cultured overnight in GM17 liquid medium containing 10 μg / mL chloramphenicol.

[00...

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Abstract

The invention provides a foot-and-mouth disease virus antigen, a gene TB1-Co1 capable of expressing the foot-and-mouth disease virus antigen, a recombinant vector comprising gene, and recombinant lactococcus lactis, and belongs to the technical field of genetic engineering vaccines. The amino acid sequence of the antigen is disclosed by SEQ ID NO:1. An animal immunity test proves that the foot-and-mouth disease virus antigen has good immunogenicity. The invention also provides a gene TB1-Co1 capable of expressing the foot-and-mouth disease virus antigen disclosed by the above scheme. The C terminal of a multi-epitope protein is blended with the targeted ligand Col of an M cell, the targeted ligand Col and a C5a receptor mutually act to deliver a model antigen connected with the ligand to an M cell, and in addition, an antigen specific immune response can be stimulated, and includes a systemic immune response and a local mucosal immune response.

Description

technical field [0001] The invention relates to the technical field of genetic engineering vaccines, in particular to a foot-and-mouth disease virus antigen, a gene TB1-Co1 expressing the foot-and-mouth disease virus antigen, its recombinant vector and recombinant Lactococcus lactis. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, febrile, highly contagious infectious disease caused by foot-and-mouth disease virus (FMDV), which mainly affects pigs, cattle, sheep and other livestock and other wild animals. Artiodactyl. The clinical feature is a vesicular eruption on the oral mucosa, hooves, and breast skin. Foot-and-mouth disease virus belongs to the genus Foot-and-Mouth Disease Virus of the Picornaviridae family. The virus genome is a single-stranded positive-strand RNA, about 8.5kb. There are 7 serotypes, and there is no cross-protection between types. Its open reading frame consists of L gene, P1 structural protein gene (including VP4, VP2, VP3, V...

Claims

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Application Information

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IPC IPC(8): C07K14/09C12N15/42C12N15/74C12N1/21A61K39/135A61P31/14C12R1/01
CPCA61K39/12A61K2039/5256A61K2039/542A61K2039/552A61P31/14C07K14/005C12N15/746C12N2770/32122C12N2770/32134
Inventor 潘丽张中旺吕建亮张富东马中元刘新生周鹏王永录张永光
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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