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Preparation of double-labeled virus-like particles (VLPs) of PRRSV

A virus-like, double-labeled technology, applied in the field of biological research, which can solve problems such as the absence of PRRSV double-labeled VLPs

Pending Publication Date: 2018-11-16
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

Although scholars at home and abroad have used the insect baculovirus expression system to prepare PRRSV VLPs, there is no report of PRRSV double-labeled VLPs.

Method used

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  • Preparation of double-labeled virus-like particles (VLPs) of PRRSV
  • Preparation of double-labeled virus-like particles (VLPs) of PRRSV
  • Preparation of double-labeled virus-like particles (VLPs) of PRRSV

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Embodiment Construction

[0062] The technical solutions of the present invention will be clearly and completely described below in conjunction with the specific embodiments of the present invention. Obviously, the described embodiments are only a part of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work shall fall within the protection scope of the present invention.

[0063] A method for preparing PRRSV double-labeled virus-like particles is:

[0064] 1. Material preparation

[0065] 1.1 Selection of strains and plasmids

[0066] Type 2 highly pathogenic PRRSV QH-08 strain and Marc-145 cells are preserved in our laboratory; Sf9 insect cell line and High Five cell line are preserved by the foot-and-mouth disease prevention and control technology innovation team of Lanzhou Veterinary Research Institute; baculovirus expression vector pFastBac TM Dual (dual promoter...

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Abstract

The invention discloses a method for preparing and identifying double-labeled virus-like particles (VLPs) of PRRSV, and relates to the field of biological research technology; a pMD18T recombinant plasmid containing GP5, M and N protein coding genes ORF5, ORF6 and ORF7 is prepared as an amplification template, a Myc tag is introduced at the end of the ORF5, a His tag is introduced into the end ofthe ORF6, a recombinant plasmid is obtained by recombination to PpH and Pp10 promoters of a binary vector, the recombinant plasmid is transferred into DH10Bac E. coli to obtain a recombinant bacmid; and the ORF7 and a pFastBacHTB vector are recombined to obtain a recombinant bacmid. The two recombinant bacmids are inoculated into SF9 cells, and Myc-GP5, His-M and His-N proteins are successfully expressed and automatically assembled into the VLPs. The method lays a foundation for studying the function of the GP5 and M and N major structural proteins and developing VLP vaccines that can distinguish between infection and immunity.

Description

Technical field [0001] The invention relates to the preparation of double-labeled virus-like particles (VLPs) of PRRSV, in particular to the technical field of biological research. Background technique [0002] Porcine Reproductive and Respiratory Syndrome (PRRS) is a viral infectious disease caused by PRRSV that seriously harms the pig industry. Mainly harmful to pregnant pigs and piglets, fattening pigs will also be sick, sows have reproductive disorders, fattening pigs respiratory infections. It can be spread through mating, drinking, mother-to-child vertical, etc. The incubation period can reach more than half a year, and it has the possibility of persistent infection and hidden infection. Once it becomes ill, it spreads quickly to the entire farm, with regional epidemics and highly contagious infections. The most effective way to prevent PRRSV infection is vaccination. However, the inactivated vaccines used clinically have the disadvantages of large immune doses, poor immu...

Claims

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Application Information

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IPC IPC(8): C12N15/866C12N15/40C07K14/08
CPCC07K14/005C12N15/86C12N2710/14043C12N2770/10022C12N2770/10023
Inventor 张杰张永光刘永生代军飞王俊丁耀忠马炳欧云文孙跃峰李茜侯谦
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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