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Gene encoding recombinant porcine circovirus type 2 Cap protein and application thereof

A porcine circovirus, protein technology, applied in applications, viral peptides, antiviral agents, etc., can solve problems such as no fusion of CPPs and Cap proteins

Active Publication Date: 2015-11-25
NANJING AGRICULTURAL UNIVERSITY +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no report on the fusion of CPPs and Cap protein

Method used

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  • Gene encoding recombinant porcine circovirus type 2 Cap protein and application thereof
  • Gene encoding recombinant porcine circovirus type 2 Cap protein and application thereof
  • Gene encoding recombinant porcine circovirus type 2 Cap protein and application thereof

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Embodiment Construction

[0047] The present invention is further described below to demonstrate the advantages of the present invention.

[0048] 1. Research methods and results

[0049] 1.1 Main reagents

[0050] PCV2SH strain virus liquid was isolated, identified and preserved in our laboratory; pET28a(+) plasmid, Escherichia coli BL21(DE3) and DH5α were purchased from Shanghai Yingjun Biotechnology Co., Ltd.; PCV2 monoclonal antibody 3E5 (preservation number CGMCC8169), rabbit anti-PCV2 Serum antibodies were prepared and stored in our laboratory. For specific methods, please refer to Zhai Shuyan, a graduate of our laboratory in 2013, master's degree thesis "Development of Porcine Circovirus Type 2 Monoclonal Antibody and Establishment of Sandwich ELISA Antigen Detection Method".

[0051] The CPH adjuvant is prepared by our laboratory. The configuration method is: weigh 50g of carbomer, add it to 9800ml of distilled water, after fully swelling, mix well, autoclave at 115°C for 30 minutes, it become...

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Abstract

The present invention provides a gene encoding a recombinant porcine circovirus type 2 Cap protein and application thereof. The present invention aims to obtain a soluble recombinant PCV2 Cap protein, and adopts a PCR method for fusion of MPG and NLS partially deleted Cap protein N-terminal gene to construct a new gene; and then the new gene is cloned into an E. coli expression vector pET28a to obtain a recombinant plasmid; the recombinant plasmid is transformed into E. coli BL21 to obtain recombinant engineering bacteria; the recombinant engineering bacteria is subjected to induced expression by IPTG to obtain the soluble recombinant PCV2 Cap protein. SDS-PAGE and Western-blot identification shows that most of the recombinant protein MrCap is present in the bacterial lysis supernatant and is soluble; and the recombinant protein MrCap has high immunogenicity and immunoreactivity, and lays foundation for the development of a PCV2 antibody detection kit and subunit vaccines.

Description

technical field [0001] The invention relates to a new gene encoding recombinant porcine circovirus type 2 (PCV2) Cap protein and the recombinant porcine circovirus type 2 Cap protein encoded by the gene, and also relates to a preparation method and application of the recombinant protein. Background technique [0002] Porcine circovirus type 2 (PCV2), considered an important pathogen associated with many swine diseases, was first discovered in western Canada in 1996 and was later reported as a new syndrome named piglet multisystemic wasting syndrome (PMWS). PCV2 belongs to the genus Circovirus of the Circoviridae family, and is one of the smallest animal viruses. Its genome is about 1.7 kb in length and is a single-stranded circular DNA virus. There are two relatively large open reading frames ORF1 and ORF2, which encode Rep and Cap proteins respectively. Rep is mainly involved in the replication of the virus, and Cap is the structural protein of the virus, which can aggrega...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/34C12N15/11C12N15/70C12N1/21C07K14/01G01N33/68A61K39/12A61P31/20
Inventor 姜平周雪晨白娟曹晶晶
Owner NANJING AGRICULTURAL UNIVERSITY
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