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Swine vesicular disease virus viral protein 1 (VP1) protein secretion expression recombinant plasmid and application thereof

A porcine vesicular disease virus and recombinant plasmid technology, applied in the field of protein expression, can solve the problems such as the inability of the recombinant protein to be secreted out of the cell, unfavorable for the purification of the recombinant protein, difficulty in protein purification, etc., so as to facilitate the extraction and purification, reduce the production cost, and improve the purification efficiency. Effect

Inactive Publication Date: 2013-03-20
SHENZHEN AUDAQUE DATA TECH
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AI Technical Summary

Problems solved by technology

[0006] At present, the use of baculovirus white expression system to express foreign proteins is basically intracellular expression, and the expression vector does not contain protein signal peptide secretion signals, so that the expressed recombinant protein cannot be secreted outside the cell. Complicated, which brings difficulties to protein purification, increases the cost, and is not conducive to the purification of recombinant proteins

Method used

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  • Swine vesicular disease virus viral protein 1 (VP1) protein secretion expression recombinant plasmid and application thereof
  • Swine vesicular disease virus viral protein 1 (VP1) protein secretion expression recombinant plasmid and application thereof
  • Swine vesicular disease virus viral protein 1 (VP1) protein secretion expression recombinant plasmid and application thereof

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Experimental program
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Embodiment

[0038] 1. Materials and methods

[0039] 1.1 Strains and plasmids

[0040] Escherichia coli (E.coli) DH5α, DH10Bac (including Bacmid and Helper plasmids), and pFastBac1 plasmids are all products of Invitrogen; pMD18-T plasmid is a product of Dalian Bao Biological Engineering Co., Ltd.; porcine vesicular disease virus nucleic acid, insect cell Sf9 Passage cells were provided by the Animal and Plant Inspection and Quarantine Technology Center of Shenzhen Entry-Exit Inspection and Quarantine Bureau.

[0041] 1.2 Main reagents

[0042] Porcine vesicular disease virus-positive and negative sera were provided by the Institute of Animal Health (IAH); the biotin-labeled anti-pig secondary antibody was produced by Abcom; SF-900 II medium and liposome II were both produced by Invitrogen.

[0043] 1.3 Construction of recombinant transposable vector

[0044] 1.3.1 Adding the vector construction of bee venom signal peptide sequence

[0045] The BamH Ⅰ restriction site sequence is added...

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Abstract

The invention relates to the protein expression field and discloses swine vesicular disease virus viral protein 1 (VP1) protein secretion expression recombinant plasmid. The recombinant plasmid is composed of a gene which expresses the swine vesicular disease virus VP1 protein and a melittin signal peptide gene sequence. The recombinant plasmid has the advantages of being capable of achieving secretion and expression of the swine vesicular disease virus VP1 protein outside an insect cell, thereby directly extracting the swine vesicular disease virus VP1 protein from cell culture fluid, effectively improving protein purification efficiency, reducing preparation cost of the swine vesicular disease virus VP1 protein and laying a solid foundation for a research based on the swine vesicular disease virus VP1 protein.

Description

technical field [0001] The present application relates to the field of protein expression, in particular to a recombinant plasmid for secretion and expression of porcine vesicular disease virus VP1 protein, construction of the recombinant plasmid, and secretion and expression of porcine vesicular disease virus VP1 protein using the recombinant plasmid. Background technique [0002] Swine vesicular disease (SVD) is an acute, febrile, contact infectious disease caused by swine vesicular disease virus (SVDV), characterized by vesicular lesions on the mouth and hooves, and its clinical Symptoms are similar to foot-and-mouth disease, vesicular stomatitis and porcine vesicular rash, and are listed as Class A infectious diseases by the World Organization for Animal Health (OIE). SVDV is a small RNA virus (Picornaviridae) enterovirus (Enterovirus). It has very similar physical and chemical characteristics, biological characteristics and serological relationship with human coxsackie...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N15/40C12N15/66C07K14/08
Inventor 曹琛福李中圣杨俊兴花群义
Owner SHENZHEN AUDAQUE DATA TECH
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