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Recombinant protein and application thereof to preparation of porcine circovirus as well as porcine reproductive and respiratory syndrome virus vaccine

A porcine circovirus and respiratory syndrome technology, applied in the field of veterinary medicine, can solve the problems of low transformation efficiency, lack of recombinant plasmids and identification methods of recombinant host bacteria, etc., to improve the efficiency of electrotransformation, improve the level of soluble expression, and achieve accurate results Effect

Active Publication Date: 2018-10-02
CHONGQING ACAD OF ANIMAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the transformation efficiency of the cloning host strain MC1061 and the expression host strain NZ9000 provided by this system is not high, lacks a convenient identification method for recombinant plasmids and recombinant host strains, and does not contain recombinant protein purification and identification tags

Method used

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  • Recombinant protein and application thereof to preparation of porcine circovirus as well as porcine reproductive and respiratory syndrome virus vaccine
  • Recombinant protein and application thereof to preparation of porcine circovirus as well as porcine reproductive and respiratory syndrome virus vaccine
  • Recombinant protein and application thereof to preparation of porcine circovirus as well as porcine reproductive and respiratory syndrome virus vaccine

Examples

Experimental program
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Effect test

Embodiment

[0053] 1. Universal primers for exogenous gene detection and sequencing in recombinant plasmid pNZ8148

[0054] 1. Design of general primers for exogenous gene detection and sequencing in recombinant plasmid pNZ8148

[0055] The specific primer pair PNZF / PNZR was designed according to the sequence of plasmid pNZ8148. The primers were located on both sides of the multiple cloning site of the plasmid, and the size of the amplified fragment was 556bp. The plasmid contains the inserted gene, and the size of the amplified fragment is about the sum of the length of the target gene and 556bp. The primer information is shown in Table 1.

[0056] Table 1 The primer pair PNZF / PNZR for detecting the inserted gene of the recombinant pNZ8148 plasmid

[0057]

[0058] 2. DNA template

[0059] Escherichia coli MC1061, TOP10 and Lactococcus lactis NZ9000 containing pNZ8148.

[0060] 3. PCR reaction and sequence analysis

[0061] Add each PCR reaction system according to Table 2, and the...

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Abstract

The invention provides a recombinant protein comprising porcine circovirus type 2 and porcine reproductive and respiratory syndrome virus epitopes, and is characterized in that the recombinant proteinhas an amino acid sequence as shown in SEQ ID NO.1. According the invention, the soluble and high-efficiency expression of the protein is realized, the recombinant protein is convenient to purify, and the purified protein is high in purity and can be applied to vaccines with better performance.

Description

technical field [0001] The invention belongs to the field of veterinary medicine, and relates to a recombinant protein and a preparation method thereof, especially for preparing porcine circovirus and / or porcine reproductive and respiratory syndrome virus vaccines. Background technique [0002] Porcine circovirus (Porcinecircovirus, PCV) was first isolated in 1974 by German scholar Tischer I et al. from multiple strains of continuously passaged pig kidney cell lines (PK15), and was named in 1982. PCV is one of the smallest animal viruses found so far. It is known that PCV has two serotypes, namely PCV1 and PCV2. PCV1 is a non-pathogenic virus and PCV2 is a pathogenic virus. The harm of porcine circovirus type 2 to the pig industry has been paid more and more attention. [0003] PCV2 is the main pathogen that causes multisystemic wasting syndrome in weaned piglets, causing progressive emaciation, cough, and dyspnea in weaned piglets. At present, PCV2 has been widespread an...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/70A61K39/12A61P31/20A61P31/14
CPCA61K39/12A61K2039/552A61K2039/70A61P31/14A61P31/20C07K14/005C07K2319/21C12N15/70C12N2750/10022C12N2750/10034C12N2770/10022C12N2770/10034
Inventor 沈克飞杨柳付利芝杨睿周雪张素辉
Owner CHONGQING ACAD OF ANIMAL SCI
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