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Probiotics cloned strain of integrated single copy functional F4 pili operon gene, construction method and application

A construction method and functional technology, applied in the field of biotechnology applications, can solve the problems of slow growth, weight loss, high mortality of piglets, etc., and achieve the effect of easy removal and improved adhesion performance

Active Publication Date: 2019-03-15
YANGZHOU UNIV
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Problems solved by technology

[0005] Piglet weaning diarrhea (PWD) and piglet edema disease (ED) are clinically common diseases in the pig industry. The main pathogens are F4 and F18 pili-positive enterotoxigenic Escherichia coli ETEC, which are infected by pili adhesion and colonization of susceptible piglets Disease, leading to high mortality of piglets, low body weight, slow growth, expensive drug treatment, etc.

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  • Probiotics cloned strain of integrated single copy functional F4 pili operon gene, construction method and application
  • Probiotics cloned strain of integrated single copy functional F4 pili operon gene, construction method and application
  • Probiotics cloned strain of integrated single copy functional F4 pili operon gene, construction method and application

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[0029] The famous probiotic Escherichia coli Nissle 1917 (EcN) is a non-pathogenic E. coli probiotic, which has been proven to give health benefits to different hosts, and E. coli Nissle1917 is a non-pathogenic strain that can give different hosts Health benefits, no known harmful effects on the host were found, and this strain is used as the main active ingredient of a probiotic product called Mutaflor in Europe, which is mainly used for the regulation of human intestinal health. The famous probiotic Escherichia coli Nissle 1917 (EcNc) was deposited in CGMCC, General Microorganism Center of China Committee for Culture Collection of Microbial Cultures, at the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the date of preservation was July 2018. 2. The deposit number is CGMCC No.16045.

[0030] In this experiment, the probiotic Nissle1917 plasmid-free clone EcNc was used as a vaccine carrier to construct...

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Abstract

The invention relates to the technical field of biology, in particular to probiotics cloned strain of integrated single copy functional F4 pili operon gene, a construction method and application. Themethod comprises the following steps: constructing pTargetT-nth / tppB::PtetF4 recombinant plasmid; and obtaining nonresistant single copy F4 pili gene integration cloned strain through construction ofthe probiotics cloned strain of the integrated single copy functional F4 pili operon gene. The probiotics cloned strain serves as probiotics live vaccine candidate strain and drug for the treatment ofpost-weaning diarrhea and hydropsy. Compared with the prior art, the probiotics cloned strain has the benefits that Nissle1917 probiotics original plasmid is removed, and resistance gene does not exist; recombinant bacteria can effectively improve the adhesion property to pig intestinal passage epithelial cell; mice subjected to oral immunization can produce immune serum namely anti-F4 pili IgG antibody; and serum of the mice subjected to oral immunization can obviously reduce adhesion to pig intestinal passage cell line by F4+ strain.

Description

technical field [0001] The invention relates to the application field of biotechnology, in particular to the chromosomal integration, stable genetic expression and display of exogenous functional F4 pili protein of bacteria. Background technique [0002] Escherichia coli Nissle1917 is a non-pathogenic probiotic strain that has no known adverse effects on the host and can give different hosts health benefits. The main active ingredient is mainly used for human intestinal health regulation. In clinical application, the probiotic Nissle1917 strain has been modified as a carrier of genetic engineering, and the probiotic has been used in the development and manufacture of vaccines, tumor treatment, health care products, diagnostic preparations and other products. The EcNc clone is a prototype wild strain of Escherichia coli Nissle1917 genetically modified to remove two cryptic plasmids pMUT1 and pMUT2 in its cell, which has the characteristics of carrying more (large) capacity h...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/90C12N15/31A61K35/741A61P31/04C12R1/19
CPCA61K2035/115A61P31/04C07K14/245C12N15/902
Inventor 朱国强区炳明金铎夏芃芃朱军徐梦娴宋浩亮梁轩杨颖朱晓芳
Owner YANGZHOU UNIV
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