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Method for preparing pig sapporo virus antigen

A virus antigen, Sapporo virus technology, applied in the field of biomedicine, can solve the problems of no vaccine and inability to obtain a large number of virus antigens

Inactive Publication Date: 2016-04-06
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

So far, there is no suitable cell line that can be used to cultivate the current epidemic strain, and the virus antigen cannot be obtained in large quantities, so there is no vaccine that can successfully prevent and treat the disease. Virus prevention is of great significance

Method used

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  • Method for preparing pig sapporo virus antigen
  • Method for preparing pig sapporo virus antigen
  • Method for preparing pig sapporo virus antigen

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Embodiment Construction

[0045] The present invention will be further described below in conjunction with the accompanying drawings. It should be noted that the present embodiment takes the technical solution as the premise, and provides a detailed implementation manner and a specific operation process, but the protection scope of the present invention is not limited to the present invention. Example.

[0046] A preparation method of pig Sapporo virus antigen, comprising the following steps:

[0047] S1 uses the RNA of SaVCH430 strain as a template, and the whole capsid protein VP1 gene is amplified by RT-PCR;

[0048] S2 clone the VP1 gene obtained in step S1 into the eukaryotic expression vector pFastBac TM In HTB, construct a positive recombinant plasmid;

[0049] S3 The positive recombinant plasmid successfully constructed in step S2 was transferred into competent cells containing DH10 baculovirus backbone, and then positive strains were screened through the medium containing three resistances of ...

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Abstract

The invention discloses a method for preparing a pig Sapporo virus antigen. The preparation method comprises the following steps: amplifying an entire capsid protein VP1 gene by taking RNA (Ribose Nucleic Acid) of an SaV CH430 plant as a template through an RT-PCR (Reverse Transcription-Polymerase Chain Reaction); cloning the entire capsid protein VP1 gene into a eukaryotic expression vector pFastBacTMHTB; transferring a positive plasmid constructed successfully into a competent cell containing a DH10 rhabdovirus skeleton; screening a cell of a recombinant rhabdovirus plasmid carrying the SaV VPI gene through three resistant culture media containing kanamycin, gentamicin and tetracycline; transfecting the obtained cell of the recombinant rhabdovirus plasmid carrying the SaV VPI gene with an sf9 cell to obtain a P1-generation rhabdovirus and a P2-generation rhabdovirus; infecting P2-generation virus liquid with the sf9 cell; harvesting a cell after 72 hours to obtain a pig Sapporo virus VP1 capsid protein and an assembled virus-like particle antigen (VLP), wherein the VLP is similar to a PoSaV natural virus in structure. By adopting the method, a PoSaV VP1 protein and a VLP antigen thereof can be obtained, and a basis is laid for the research of subunit vaccines.

Description

technical field [0001] The art relates to the field of biomedicine, in particular to a method for preparing a pig Sapporo virus antigen. Background technique [0002] In piglets, SaV causes severe viral gastroenteritis, especially in concentrated breeding areas, once the clinical symptoms appear, it is more serious, and it will also cause certain economic losses and pose a potential threat to public health. Up to now, there is no suitable cell line that can be used to cultivate the current circulating strain, and the virus antigen cannot be obtained in large quantities, and there is no vaccine to successfully prevent and control the disease. Virus control is very important. To date, several VLP vaccines have been commercialized and licensed commercially. This means that the development of new vaccines is particularly urgent. SUMMARY OF THE INVENTION [0003] In view of the deficiencies of the prior art, the present invention aims to provide a preparation method of swine...

Claims

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Application Information

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IPC IPC(8): C12N15/866C07K14/08C12N7/04
CPCC07K14/005C12N7/00C12N2770/16022C12N2770/16023
Inventor 兰喜
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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