gg and TK gene-deleted recombinant infectious bovine rhinotracheitis virus and application

A rhinotracheitis virus and infectious technology, applied in the field of animal genetic engineering, can solve the problems of high serological positive rate and harm to the development of aquaculture industry, etc.

Active Publication Date: 2010-09-01
HUAZHONG AGRI UNIV
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Problems solved by technology

However, the serological positive rate of IBRV in my country is very high (B.F.Yan, Serological survey of bovine herpesvirus type 1 infection in China, veterinary microbiogy, 2007, 127, 136-141). Bovine infectious rhinotracheitis is widely spread worldwide. , seriously endanger the development of the breeding industry and our country has no countermeasures, so it is very meaningful to be able to develop a vaccine suitable for my country's national conditions

Method used

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  • gg and TK gene-deleted recombinant infectious bovine rhinotracheitis virus and application
  • gg and TK gene-deleted recombinant infectious bovine rhinotracheitis virus and application
  • gg and TK gene-deleted recombinant infectious bovine rhinotracheitis virus and application

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Embodiment 1

[0048] 1. Primer Design

[0049] Plasmid pEGFP-C1 (purchased from Becton, Dickison and Company) was double-digested with Bam H I and Bgl II, and T4 ligase was used as a template to amplify the marker gene EGFP, and the restriction sites were introduced as Kpn I, Bam H I , the fragment size is about 1900bp; according to the full-length nucleotide sequence of the bovine infectious rhinotracheitis virus genome published on GenBank (GenBank accession number: AJ004801), two pairs of primers were designed, and IBRV precipitation DNA was used as a template to amplify respectively The upstream and downstream homology arms of the gG gene, the designed enzyme cutting sites are HindIII, KpnI, the fragment size is about 800bp, 1000 and intermediate product PCR identification primers of the present invention (primers are synthesized by Shanghai Sangong Bioengineering Technology Co., Ltd.) Respectively as follows (the following primers can be identified in the sequence of SEQ ID NO: 1-3 des...

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Abstract

The invention belongs to the fields of animal virology and animal genetically engineered technology, and in particular relates to a gG and TK gene-deleted recombinant infectious bovine rhinotracheitis virus, a genetic engineering vaccine and application. The recombinant infectious bovine rhinotracheitis virus strain IBRV delta gG / delta TK prepared by the invention is preserved in China Center for Type Culture Collection, with the preservation number of CCTCC No: V200915; and gG and TK genes are deleted in the strain, and the strain does not comprise foreign genes. An intermediate strain, namely recombinant infectious bovine rhinotracheitis virus GAZH-2009-01 is preserved in China Center for Type Culture Collection, with the preservation number of CCTCC No: V200910; and gG and TK genes are deleted in the strain and the strain comprises reporter gene EGFP. The invention also discloses application of the recombinant infectious bovine rhinotracheitis virus IBRV delta gG / delta TK in preparing the genetic engineering vaccine for the infectious bovine rhinotracheitis and preventing and controlling infectious bovine rhinotracheitis virus diseases.

Description

technical field [0001] The invention belongs to the technical field of animal genetic engineering, and specifically relates to a recombinant bovine infectious rhinotracheitis virus with double gene deletion. The virus lacks gG and TK genes, and the virulence of the virus strain is reduced but the immunogenicity is not affected. The present invention The invention also includes using the recombinant virus to prepare bovine infectious rhinotracheitis virus vaccine and the application of the vaccine. Background technique [0002] Infectious Bovine Rhinotracheitis (IBR), also known as "necrotizing rhinitis" or "red nose", is caused by Infectious Bovine Rhinotracheitis viruses (IBRV) in cattle. An acute, febrile, contact infectious disease. The danger of this disease is that after the virus invades the body of cattle, it can hide in a certain part, resulting in persistent infection. The sick cattle are infected with the virus for a long time or even for life. When the body's resi...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/265A61P31/20C12N7/01C12R1/93
Inventor 郭爱珍张敏敏刘正飞付书林陈焕春
Owner HUAZHONG AGRI UNIV
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