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Genetically engineered bacteria strain expressing porcine transmissible gastroenteritis virus

A genetically engineered bacteria and infectious technology, applied in the field of expressing porcine infectious gastroenteritis virus genetically engineered bacteria strains, can solve the problems of vaccine absorption, inability to settle in intestinal mucosa, and the harm of bacteria and viruses to the human body.

Inactive Publication Date: 2013-04-24
SHANDONG SINDER TECH
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Problems solved by technology

[0003] However, the existing vaccines generally need to be injected. Routine injection of vaccines will cause stress and problems in vaccine absorption. When immunized orally, the immunogen may be degraded or inactivated before reaching the small intestinal mucosa. This purpose can be achieved as a live carrier, but bacterial viruses are harmful to the human body and cannot settle in the intestinal mucosa

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  • Genetically engineered bacteria strain expressing porcine transmissible gastroenteritis virus
  • Genetically engineered bacteria strain expressing porcine transmissible gastroenteritis virus
  • Genetically engineered bacteria strain expressing porcine transmissible gastroenteritis virus

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preparation example Construction

[0009] see figure 1 As shown, a preparation method of a porcine epidemic diarrhea virus genetically engineered subunit oral vaccine of the present invention comprises the following steps:

[0010] 1. Extraction of total RNA

[0011] Take 250 μl of the treated TGEV cytotoxic suspension and add them to a sterile RNase-free centrifuge tube, then add 750 μl of TRIzol LS Reagent, invert and mix, and then let stand at 4°C for 5 minutes. Add 200 μl of pre-cooled chloroform (stored at 4°C), shake until fully emulsified and uniform, and let stand at 4°C for 15 minutes. Centrifuge at 4°C and 12000 r / min for 15 minutes, take out the centrifuge tube, gently suck about 450 μl of the upper aqueous phase and transfer it into another centrifuge tube. Add an equal volume of cold isopropanol (stored at -20°C), invert and mix evenly, and leave at -20°C for 30 minutes. Centrifuge at 12,000 r / min at 4°C for 10 minutes, take out the centrifuge tube, discard the supernatant, add 1 ml of 75% cold ...

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Abstract

The invention discloses a genetically engineered bacteria strain expressing a porcine transmissible gastroenteritis virus. The strain is Lactococcus lactis MG1363 / pMG36e-S preserved in China center for type culture collection, with a preservation number of CCTCC M 2012355. Lactococcus lactis is a common bacteria in intestinal tracts of human and majority of animals, has characteristics of bacterial resistance and diarrhoea resistance, and can be widely used for researches of viable cell oral vaccines. The recombinant Lactococcus lactis MG1363 / pMG36e-S constructed in the invention can stably and reliably express S protein of active TGEV, can be used for producing genetically engineering subunit vaccines and genetically engineered live vector vaccines, provides a new method for preventing TGE, and is relatively low in cost.

Description

technical field [0001] The invention relates to a gene engineering bacterial strain expressing porcine transmissible gastroenteritis virus. Background technique [0002] Porcine transmissible gastroenteritis (TGE) is a digestive tract infectious disease characterized by piglet vomiting, severe diarrhea and high lethality caused by porcine transmissible gastroenteritis virus (TGEV) of the Coronaviridae family. . The disease is one of the important diseases of early death of piglets in my country and various pig-raising countries in the world. Vaccination is the main measure to prevent this disease. Practice has proved that the secretory antibody (sIgA) produced by intestinal mucosal immunization is an effective antibody against TGEV infection, while other serum antibodies produced by oral immunization, such as IgG and IgM, are not ideal for the immune protection of this disease . Therefore, it is of great significance to choose a carrier system that is safe and non-toxic,...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/50C07K14/17C12R1/46
Inventor 李朝阳李明义石乔
Owner SHANDONG SINDER TECH
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