Method for purifying recombinant porcine circovirus 2 type Cap protein

A porcine circovirus, protein technology, applied in chemical instruments and methods, microorganism-based methods, biochemical equipment and methods, etc., can solve problems such as the ineffective expression of Cap genes

Pending Publication Date: 2017-03-29
ZHONGCHONG XINNUO BIOTECH TAIZHOU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no report on the market to express and produce PCV-2 subunit vaccines with Escherichia coli (Escherichia coli) (hereinafter referred to as Escherichia coli in the present invention). Positioning signals or optimized codons can express

Method used

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  • Method for purifying recombinant porcine circovirus 2 type Cap protein
  • Method for purifying recombinant porcine circovirus 2 type Cap protein
  • Method for purifying recombinant porcine circovirus 2 type Cap protein

Examples

Experimental program
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Effect test

Embodiment 1

[0047] ——Preparation of recombinant porcine circovirus type 2 Cap protein

[0048] Source of biological material:

[0049] 1. pET-30a vector: purchased from Novagen, USA, and stored in our laboratory.

[0050] 2. Escherichia coli DH5a: purchased from BD Biosciences Clontech in the United States, and kept in our laboratory.

[0051] 3. BLR Escherichia coli: purchased from Novagen in the United States and stored in our laboratory.

[0052] The specific operation steps are as follows:

[0053] 1. Construction of pET-ELP vector

[0054] (1) Synthesize the ELP elastin-like polypeptide coding sequence (SEQ ID No.1), introduce the Nde I restriction site at the 5' end, introduce the Sac I restriction site at the 3' end, and send the sequence to Nanjing Jinruisi Biotechnology Co., Ltd. Co., Ltd., cloned in the pUC57 plasmid vector, SEQ ID No.1 is shown as follows:

[0055]

[0056]

[0057] (2) Digest the pUC57-ELP vector with restriction enzymes Nde I and Sac I, and after s...

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Abstract

The invention relates to a method for purifying recombinant porcine circovirus 2 type Cap protein. The recombinant porcine circovirus 2 type Cap protein is composed of ELP type elastin polypeptide genes and PCV-ORF2 genes. A preparation method of the purifying recombinant porcine circovirus 2 type Cap protein comprises the steps of expression vector construction and fusion protein expressing and purifying. Compared with an existing porcine circovirus vaccine purifying method, the method for purifying the recombinant porcine circovirus 2 type Cap protein has the advantages that preparation is easy and convenient, purifying is easy, and the price is low, and is suitable for large-scale production operation, and a foundation is provided for preparation, purification and application of subsequent recombinant porcine circovirus subunit vaccines.

Description

technical field [0001] The invention relates to a method for purifying recombinant porcine circovirus type 2 Cap protein. It belongs to the field of veterinary biological products. Background technique [0002] Porcine circovirus (porcine circovirus, PCV) is a member of the genus Circovirus in the family Porcine Circoviridae. PCV is divided into two genotypes, PCV1 and PCV2. PCV1 is non-pathogenic, while PVC2 is the pathogenic factor of porcine post-weaning multisystemic wasting syndrome (PMWS). The virus has a high infection rate in pig farms in my country, and the average positive rate of pig herds is generally greater than 20%, and some even exceed 50%. Considerable economic losses have been caused to large-scale pig farms. The prevention and control of this disease mainly depends on vaccines. Currently commercialized PCV2 vaccines include PCV2 whole virus inactivated vaccine, PCV1-PCV2 chimeric virus inactivated vaccine and baculovirus expressed Cap protein inactivate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/70C12N1/21C12R1/19
CPCC07K14/005C07K14/78C07K2319/00C12N15/70C12N2750/10022C12N2800/101C12N2800/22
Inventor 张传明汤眀元李光亚
Owner ZHONGCHONG XINNUO BIOTECH TAIZHOU CO LTD
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