Vaccine for preventing swine transmissible gastroenteritis

An infectious and gastroenteritis technology, applied in the field of genetic engineering, can solve problems such as poor immunogenicity, and achieve the effects of excellent clinical application prospects, good safety and strong protection.

Active Publication Date: 2015-07-01
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, the patent application whose publication number is CN 103194471A discloses a bigenic lactic acid bacteria vaccine comprising a segment of the porcine transmissible gastroenteritis virus S gene and an N gene, but the immunogenicity is not good

Method used

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  • Vaccine for preventing swine transmissible gastroenteritis
  • Vaccine for preventing swine transmissible gastroenteritis
  • Vaccine for preventing swine transmissible gastroenteritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1 Construction of recombinant bacteria of the present invention

[0068] 1. Preparation method

[0069] 1 Construction of double-promoter eukaryotic plasmid

[0070] 1.1 RT-PCR amplification of S gene and N gene

[0071] According to the S gene sequence of TGEV, a pair of primers were designed to amplify the 5’-1kb antigenic region of the S gene (named Sa, including four antigenic sites A, B, C, and D), and the upstream primer S1: 5’-CG GCTAGC ACCATGGAAAAACTATTTG-3', introduce restriction site NheI; downstream primer S2: 5'-CC AAGCTT TTAGCCACTAAGTAGCGTC-3', introduce restriction site HindIII. Design a pair of primers according to the N gene sequence, the expected amplification length is, upstream primer N1: 5'-CG GAATTC AAAATGGCCAACCAGGGAC-3', introduce restriction site EcoRI; downstream primer N2: 5'-CC CTCGAG AAGACGAGCATCTC-3', the restriction site XhoI was introduced.

[0072] Take TGEV cell culture virus liquid, extract viral RNA according to th...

experiment example 1

[0130] Experimental example 1 Characteristic identification and protective detection of recombinant attenuated Salmonella of the present invention

[0131] 1. Experimental method

[0132] Get the recombinant Salmonella SL7207 (pVAX-S) prepared in Example 1 499-650 ), according to the following method for identification:

[0133] 1 Identification of biological characteristics of strain SL7207 (pVAXD-N-Sa)

[0134] 1.1 Microscopic examination of Gram staining of bacterial strain SL7207 (pVAXD-N-Sa) and control strain

[0135]SL7207(pVAXD-N-Sa) and control strains SL7207(pVAX-S), SL7207(pVAX-N), SL7207(pVAX-S-N) and SL7207(pVAXD) were respectively streaked on LB plates containing 100 μg / mL Kan, Cultivate for 16 hours, pick a single colony, inoculate 5 mL of LB liquid medium containing 100 μg / mL Kan, culture at 37 °C, shake at 300 r / min until the OD600 is about 0.6, take the bacterial liquid for Gram staining and microscopic examination, observe and record the bacteria form. ...

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Abstract

The invention discloses an application of recombinant salmonella containing nucleotide sequences shown in SEQ ID NO: 1 and SEQ ID NO: 2 in preparation for a medicine for preventing swine transmissible gastroenteritis, and further discloses a vaccine for preventing swine transmissible gastroenteritis, which is prepared from recombinant salmonella containing nucleotide sequences shown in SEQ ID NO: 1 and SEQ ID NO: 2, and pharmaceutically acceptable auxiliary materials or carriers. The vaccine disclosed by the invention is capable of inducibly producing two specific antibodies with regard to the S-protein and the N-protein of swine transmissible gastroenteritis virus (TGEV) simultaneously, has a double-gene immunologic function, a more comprehensive immunologic effect and a high level of the induced antibodies, as well as is capable of providing a more comprehensive protection function for swine transmissible gastroenteritis and excellent in clinical application prospect.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a vaccine for preventing porcine transmissible gastroenteritis. Background technique [0002] Porcine transmissible gastroenteritis (TGE) is a highly contagious intestinal infectious disease of pigs caused by porcine transmissible gastroenteritis virus (TGEV). , diarrhea and dehydration death, pigs of different breeds and ages are susceptible to the disease, especially the mortality rate of piglets within 2 weeks of age can be as high as 100%. Since 2010, intractable diarrhea in piglets has occurred in a large area of ​​pig herds in my country, and TGEV is one of the main pathogens frequently detected. [0003] At present, most of them are traditional inactivated vaccines and attenuated vaccines, but they have the risk of strong virulence and loose poison, which needs to be improved. In order to develop a porcine epidemic diarrhea vaccine that is safer, more efficient, less e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/225A61P31/14C12N1/21C12R1/42
Inventor 黄小波曹三杰张丹文心田张鑫淼文翼平伍锐张小会梁恩涛
Owner SICHUAN AGRI UNIV
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