Method for assembling foot and mouth disease virus hollow capsid in insect with acidproof improvement

A foot-and-mouth disease virus, acid-resistant technology, applied in viral peptides, antiviral agents, botanical equipment and methods, etc., can solve the problem of difficult to assemble FMDV empty capsid protein and other problems

Active Publication Date: 2008-09-24
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, since the pH value of the culture medium of insect cells is around 6.3, it is difficult to use the baculovirus expression system to assemble stable FMDV empty capsid proteins in insect cells.

Method used

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  • Method for assembling foot and mouth disease virus hollow capsid in insect with acidproof improvement
  • Method for assembling foot and mouth disease virus hollow capsid in insect with acidproof improvement
  • Method for assembling foot and mouth disease virus hollow capsid in insect with acidproof improvement

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Experimental program
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Effect test

Embodiment 1

[0047] Embodiment 1, the assembly of foot-and-mouth disease virus empty capsid in insect cell

[0048] 1. Cloning of the target gene

[0049] 1. RNA extraction and reverse transcription

[0050] According to the instruction manual of RNeasy Mini Kit, respectively from the Asia I / JS / 05 strain (the national foot-and-mouth disease reference laboratory of the foot-and-mouth disease virus preservation institution designated by the Ministry of Agriculture, references: Arch Virol, 2007, 152: 1699-1708 Comparisons of the complete Genomes of two Chinese isolates of a recent foot-and-mouth disease type Asia 1 virus. GenBank serial number: EF149009), O / Akesu / 58 and A / XJ / KT / 58 strains (the national foot-and-mouth disease virus depository designated by the Ministry of Agriculture) Foot-and-mouth disease reference laboratory, references: Arch Virol, 2007, 152: 2079-2085 The complete genome sequence of foot-and-mouth disease virus O / Akseu / 58 strain and its some molecular characteristics. Ge...

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Abstract

The present invention discloses a method for assembling foot-and-mouth disease virus empty capsids in insect cells via the alteration of acid-resistance. The method for assembling foot-and-mouth disease virus empty capsids in insect cells includes the following steps: (1) the altered P12A gene and the non-structural protein gene 3C of foot-and-mouth disease virus are introduced into bacteria via baculovirus vectors for recombination to produce recombinant rhabdovirus A; (2) the DNA of the recombinant rhabdovirus A is used to transfect the insect cells, so that the foot-and-mouth disease virus empty capsids are obtained. The method assembles the integral foot-and-mouth disease virus empty capsids in the insect cells for the first time, lays a foundation for the research and the development of gene-engineered subunit vaccines and novel diagnostic reagents.

Description

technical field [0001] The invention relates to a method for assembling empty capsids of foot-and-mouth disease virus in insects through acid-resistant modification. Background technique [0002] Foot-and-mouth disease (FMD) is an acute and severe infectious disease of cloven-hoofed animals. It ranks first among Class A animal infectious diseases. All countries in the world have invested huge sums of money in the prevention and treatment of this disease. Traditional vaccines have deficiencies in the prevention and treatment of foot-and-mouth disease, mainly in the unsafe use of vaccines and the difficulty in differential diagnosis of infected animals and immunized animals; at the same time, in the production process of inactivated vaccines, a large number of natural viruses need to be strictly controlled. Facilities to prevent the spread of the virus, however, still have the possibility of virus escape and incomplete virus inactivation. The outbreak of foot-and-mouth diseas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/09C12N15/42C12N15/63C12N5/10C12N15/866A61K39/135A61P31/14G01N33/569
Inventor 刘在新曹轶梅卢曾军孙普李平花郭建宏谢庆阁
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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