Recombinant bacillus subtilis for expressing S protein of transmissible gastroenteritis of swine virus

A Bacillus subtilis, infectious technology, applied in the field of biotechnology genetic engineering, can solve the problems of uncontrolled expression, pathogenicity, and attenuated bacteria returning to strong virulence

Inactive Publication Date: 2016-08-24
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

Firstly, the attenuated bacteria may have the risk of strong virulence; secondly, the plasmid DNA may be integrated into the host cell genome, which will cause the disorder of genes regulating cell growth, the uncontrolled expression of proto-oncogenes and tumor suppressor genes, somatic cell carcinogenesis, A series of issues such as cell transformation
In addition, attenuated bacteria may be potentially pathogenic in some elderly, infirm, and disabled individuals

Method used

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  • Recombinant bacillus subtilis for expressing S protein of transmissible gastroenteritis of swine virus
  • Recombinant bacillus subtilis for expressing S protein of transmissible gastroenteritis of swine virus
  • Recombinant bacillus subtilis for expressing S protein of transmissible gastroenteritis of swine virus

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Embodiment Construction

[0069] 1 Construction of integrated expression plasmid pIncotGSR

[0070] The methods used in the following examples are conventional methods unless otherwise specified. The specific materials and reagents involved are as follows: Bacillus subtilis WB800 strain was donated by Gao Xuewen, School of Plant Protection, Nanjing Agricultural University. For literature, see: Wu, H., Wang, S., Qiao, J., Liu, J., Zhan, J. , Gao, X., 2009.Expression of HpaG Xooc protein in Bacillus subtilis and its biological functions. Journal of Microbiology and Biotechnology 19, 194-203;

[0071] Porcine transmissible gastroenteritis virus SHXB strain was kindly donated by He Kongwang, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences. For literature, see: Weiwei, H., Qinghua, Y., Liqi, Z., Haofei, L., Shanshan, Z., Qi , G., Kongwang, H., Qian, Y. 2014. Complete genomic sequence of the coronavirus transmissible gastroenteritis virus SHXB isolated in China. Archives of viro...

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Abstract

The invention relates to recombinant bacillus subtilis for expressing S protein of transmissible gastroenteritis of swine virus (TGEV), and belongs to the fields of biotechnology and genetic engineering. According to the recombinant bacillus subtilis disclosed by the invention, a TGEV S protein recombinant bacillus subtilis integrated expression plasmid pIncotGSR is successfully constructed, and the expression plasmid is successfully transformed into bacillus subtilis WB800 through electric shock. Based upon Western-blot verification, the TGEV S protein can be successfully expressed in the recombinant bacillus subtilis. An effective TGEV specific immune response level can effectively generate in the body of a one-month-old piglet through stimulation of oral immunization. The recombinant bacillus subtilis is expected to be developed as a genetic engineering oral vaccine for preventing transmissible gastroenteritis of swine.

Description

1. Technical field [0001] The invention relates to a recombinant bacillus subtilis expressing the S protein of porcine transmissible gastroenteritis virus, belonging to the field of biotechnology genetic engineering. Specifically including: the construction of the integrated expression plasmid pIncotGSR of porcine transmissible gastroenteritis virus S protein, the expression and identification of porcine transmissible gastroenteritis virus S protein, and the ability of recombinant Bacillus subtilis to stimulate one-month-old piglets to produce effective TGEV-specific level of immune response. 2. Background technology [0002] 1. S protein (S), one of the main structural proteins of porcine transmissible gastroenteritis virus (TGEV) [0003] Transmissible gastroenteritis of swine (TGE) is a highly contagious intestinal infectious disease caused by porcine transmissible gastroenteritis of swine virus (TGEV), which can cause Piglet vomiting, severe diarrhea, dehydration and h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/75C12N15/66C12N1/21A61K39/225A61P31/14C12R1/125
Inventor 杨倩牟春晓朱立麒林建
Owner NANJING AGRICULTURAL UNIVERSITY
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