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Porcine circovirus (PCV) type 2 Cap protein and preparation method thereof

A porcine circovirus and protein technology, applied in the field of genetic engineering, can solve the problems of long test period, large workload and low success rate, and achieve the effects of shortening the test period, low cost and simple operation.

Inactive Publication Date: 2013-02-06
PU LIKE BIO ENG
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  • Abstract
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  • Claims
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Problems solved by technology

However, this method requires co-transfection of BmNPV and AcNPV and plaque screening to obtain recombinant silkworm nuclear polyhedrosis virus containing PCV2 Cap protein, and the probability of homologous recombination between BmNPV and AcNPV is low, and there is work on plaque screening. The disadvantages of large amount and low success rate; in addition, plaque screening needs to cultivate silkworm cells, requires TC-100 medium containing fetal bovine serum (FBS), and the culture period is long (generally 7-8 days)
Therefore, this plaque screening method has the disadvantages of heavy workload, high test cost, and long test period.

Method used

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  • Porcine circovirus (PCV) type 2 Cap protein and preparation method thereof
  • Porcine circovirus (PCV) type 2 Cap protein and preparation method thereof
  • Porcine circovirus (PCV) type 2 Cap protein and preparation method thereof

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Embodiment Construction

[0031] The insect cells used in the embodiment of the present invention are Sf9 cells (Spodoptera frugiperda (Spodoptera frugiperda) ovary cells), purchased from Invitrogen. Other types of insect cells, such as silkworm embryo cells, silkworm ovary cells (BmN), etc., can use the method of the present invention to produce porcine circovirus type 2 Cap protein as long as they can infect the transfection virus of the present invention.

[0032] The silkworm variety used in the embodiment of the present invention is Dazao, and other silkworm varieties consistent with it, as described silkworm varieties include European species: Luobendihong, Baohuang, Hungary; Chinese species: Qiansanmian, Datuanyuan, Ankang, Lan5; Japanese species: Ri110, Dongfei, Su16, Su14, YingHXZ, Haoyue, etc., can also use the method of the present invention to produce porcine circovirus type 2 Cap protein.

[0033] The porcine circovirus type 2 Cap protein of the present invention can use pharmaceutical car...

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Abstract

The invention provides a method for producing PCV2 Cap protein by using a silkworm bioreactor, and a Cap protein product. The method comprises the steps that: a coding gene sequence of the PCV2 Cap protein is cloned to a recombinant baculovirus; the recombinant insect baculovirus is used in direct injection infection of silkworm larva or silkworm pupa; and PCV2 Cap protein is recovered, separated, and purified. The method provided by the invention has the characteristics of simple operation and short period. A difficult and low-success-rate step which is virus homologous recombination plaque screening is avoided, such that an experiment period is greatly shortened. The operation is simple, the cost is low, and a post-extraction method is simple. The method provided by the invention is suitable for industrialized productions.

Description

technical field [0001] The invention relates to a porcine circovirus type 2 Cap protein and a preparation method thereof, in particular to a porcine circovirus type 2 Cap protein produced and prepared in silkworms by means of genetic engineering and a method thereof, belonging to the field of genetic engineering. Background technique [0002] Porcine circovirus (Porcine circovirus, PCV) belongs to the genus Circovirus, is a icosymetric, covalently closed, circular, single-stranded DNA virus. According to the pathogenicity, antigenicity and nucleotide sequence of PCV, PCV can be divided into porcine circovirus type 1 (PCV1) and porcine circovirus type 2 (PCV2). Among them, PCV1 is not pathogenic to pigs, and PCV2 is very harmful to the clinic. It can cause postweaning multisystemic wasting syndrome (Postweaning multisystemic wasting syndrome, PMWS), porcine dermatitis nephrotic syndrome (PDNS), and porcine respiratory disease syndrome. The outbreak of various diseases such a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/34C12N15/866C12N7/01C07K14/01
Inventor 张许科孙进忠白朝勇
Owner PU LIKE BIO ENG
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