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Recombinant baculovirus strain of porcine circovirus type 2 Cap protein expression, construction method and application thereof

A technology of recombinant baculovirus and porcine circovirus, which can be applied in application, antiviral agent, virus antigen component, etc., can solve the problems of difficult purification, low toxicity of virus culture and difficult research work.

Inactive Publication Date: 2009-02-04
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Because the virus-infected cells do not show cytopathic changes, the virus culture has low toxicity and is not easy to purify, which brings certain difficulties to the research work.

Method used

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  • Recombinant baculovirus strain of porcine circovirus type 2 Cap protein expression, construction method and application thereof
  • Recombinant baculovirus strain of porcine circovirus type 2 Cap protein expression, construction method and application thereof
  • Recombinant baculovirus strain of porcine circovirus type 2 Cap protein expression, construction method and application thereof

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Embodiment 1

[0015] Embodiment 1: Construction of recombinant baculovirus strain (rBac / PCV2Cap) of the present invention and expression of recombinant protein

[0016] 1 Materials and methods

[0017] 1.1 Viruses, cells and antiserum

[0018]Porcine circovirus type 2 strain (PCV2 / LG strain): It is obtained from the organs of sick pigs with clinical symptoms of PMWS in a local pig farm, isolated from a pig kidney cell line that does not contain PCV1, and continuously passed for 29 generations . RPMI 1640 culture medium (Gibco) was used for cell culture, supplemented with 10% fetal bovine serum, 100 U / ml penicillin and 100 μg / ml streptomycin. Carry out cell culture according to routine method, adopt synchronous method to inoculate virus, when reaching 80% monolayer, according to literature (TischerI, Mields W, Wolff D, et al.Studies on epidemiology and pathogenicity of porcine circovirus [J]. 91:271-276.) method for D-glucosamine treatment. Preparation of antiserum: A 30-day-old virus-an...

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Abstract

The present invention discloses a recombinant baculovirus strain rBac / PCV2Cap (microorganism preservation number: CGMCC NO.2083) efficiently expressing Porcine circovirus type 2 Cap protein and applications thereof. The recombinant baculovirus strain rBac / PCV2Cap constructed by the present invention can efficiently express recombinant PCV2-Cap protein in insect cells, and the expressed recombinant Cap protein, which has good immunocompetence and antigenicity, can serve as a subunit vaccine used to prevent the related plague caused by Porcine circovirus type 2 infection as well as a detecting and diagnostic antigen for Porcine circovirus type 2 serum antibody.

Description

technical field [0001] The invention relates to a method for constructing a recombinant baculovirus strain, in particular to the expression and application of a porcine circovirus type 2 recombinant Cap protein; the invention also relates to the recombinant expression used to construct the porcine circovirus type 2 recombinant strain The carrier and the application of the recombinant virus strain in the diagnosis and prevention of porcine circovirus belong to the field of veterinary biotechnology. Background technique [0002] Porcine circovirus type 2 (PCV2) is the causative agent of Postweaning multsystemic wasting syndrome (PMWS), which causes progressive emaciation, cough, dyspnea, diarrhea, and pale skin in weaned piglets Or yellow staining, lymphadenopathy, kidney gray-white edema and other symptoms (Allan G M, Ellis J A. Porcine circoviruses: A review [J]. J Vet Diagn Invest, 2000, 12: 3-14). The disease was discovered in Canada for the first time in 1991, and it has...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/866A61K39/12A61P31/12G01N33/53
CPCY02A50/30
Inventor 刘长明陆月华张超范危艳武张朝霞袁婧童光志
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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