Infectious bovine rhinotracheitis virus recombinant strain, construction method and use thereof

A rhinotracheitis virus, infectious technology, applied in the field of genetic engineering, can solve problems such as vaccines not being put into use

Active Publication Date: 2009-01-28
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
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Problems solved by technology

But none of these vaccines have been used

Method used

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Embodiment Construction

[0029] The present invention is further described by the following examples. It should be understood that these examples are only for the purpose of illustration, and in no way limit the protection scope of the present invention.

[0030] The present invention uses the IBRV Bartha-Nu / 67 attenuated strain introduced from Hungary to construct the IBRV gE gene deletion strain IBRV46, and the 1134 bp of the gE gene encoding nucleotide including the initiation codon ATG of the strain has been successfully deleted (The full length of gE gene is 1728bp). The construction process of IBRV gE gene deletion strain IBRV46 is as follows:

[0031] 1. Preparation of IBRV Genomic DNA Template

[0032] IBRV genomic DNA was extracted with reference to the method reported by Pignatti et al., with slight improvements. Inoculate bovine fetal turbinate cell monolayer with IBRVBartha-Nu / 67. After 80% of the cells have lesions, wash the cell monolayer twice with pre-cooled 0.01M pH7.4PBS, scrape th...

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Abstract

The invention discloses an IBRV recombinant transfer vector and a constructing method thereof, and also discloses a recombinant strain IBRV46 obtained by the recombinant transfer vector and an IBRV genome after being co-transfect. A gE gene in the recombinant transfer vector, including the 1134bp comprising an initiation codon ATG is deleted, the position where the gE gene is deleted is inserted with a LacZ expression cassette. The recombinant strain of the invention can be used for preparing a vaccine for preventing and curing IBRV and can also be used for preparing diagnostic reagent. According to the conventional molecular biological method or serological method, the strain-deleted infection of the gE gene is distinguished from the wild strain infection. Additionally, the strain can be further remodeled or the nonessential genes of the strain are deleted, therefore, a mutated strain is obtained, and the vaccine, etc., can be further prepared.

Description

technical field [0001] The invention relates to a recombinant virus strain, in particular to a bovine infectious rhinotracheitis virus recombinant strain and a method for constructing the recombinant virus strain. The invention also relates to the application of the recombinant virus strain as a bovine infectious rhinotracheitis virus vaccine, a diagnostic reagent or as a virus vector for exogenous gene expression, which belongs to the field of genetic engineering. Background technique [0002] Infectious bovine rhinotracheitis virus (IBRV), also known as bovine herpesvirus type 1, is an acute contact infectious disease characterized by upper respiratory tract inflammation in cattle. Clinical manifestations of various disease types, such as upper respiratory tract mucosal inflammation, pustular vulvovaginitis, balanitis, conjunctivitis, young cattle meningoencephalitis, mastitis, abortion and so on. IBRV grows well in a variety of bovine cell cultures such as turbinate, tra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/86C12N15/38C12N7/00A61K39/245G01N33/571
Inventor 薛飞朱远茂童光志王延辉
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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