Method for preparing porcine circovirus 2 open reading frame 2 (ORF2) protein

A circovirus and ORF2 technology, which is applied in the field of preparation of porcine type II circovirus ORF2 protein, can solve the problems of high price, high expression cost, lack of production inspection methods and standards, etc., so as to reduce production costs and avoid protein denaturation. Effect

Inactive Publication Date: 2013-03-27
SHANDONG SINDER TECH
View PDF2 Cites 11 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Porcine circovirus type II (PCV2) cell culture inactivated vaccine yield is low and the country lacks a unified production and inspection method and standard for PCV2 vaccine. These problems have become bottlenecks that limit the development and application of this type of product; new vaccines such as DNA vaccine , the immune efficiency has yet to be confirmed, and subunit vaccines have been marketed a

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for preparing porcine circovirus 2 open reading frame 2 (ORF2) protein
  • Method for preparing porcine circovirus 2 open reading frame 2 (ORF2) protein
  • Method for preparing porcine circovirus 2 open reading frame 2 (ORF2) protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0022] see figure 1 and figure 2 Shown, the preparation method of a kind of porcine type II circovirus ORF2 protein of the present invention comprises the following steps:

[0023] 1. Sequence Synthesis

[0024] Through sequence alignment, the nucleotide sequence is changed under the premise of ensuring that the encoded amino acids are the same, the rare codons in the sequence are replaced with E. coli preferred codons, and EcoR I and Not I restrictions are added to the two ends of the sequence respectively Restriction site, synthesize cap gene, connect to PES vector, digest according to the set restriction endonuclease, clone the target fragment into pMD18-T plasmid vector, transform competent DH5a cells, pick positive colonies , A small amount of plasmid was extracted with OMEGA kit, and the positive plasmid was identified by enzyme digestion.

[0025] 2. Construction and identification of prokaryotic expression vector

[0026] The plasmid pES-Cap was digested by EcoR ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method for preparing a porcine circovirus 2 open reading frame 2 (ORF2) protein. A Cap gene of an encoding porcine circovirus 2 (PCV2) nucleocapsid protein is connected to a pET-28a(+) plasmid to build an expression plasmid pET-28-Cap, and a BL21(DE3) competent cell is converted to obtain a genetic engineering strain pET-28-Cap/BL21. Induction expression and detection prove that the recombinant protein has good reactionogenicity; furthermore, the protein can be used for diagnosing antigen or preparing subunit vaccines; and the product can be expressed in Escherichia coli in a soluble manner, the expression quantity is large and a complicated process of albuminous degeneration repeatability is avoided, and the loss and production cost of the protein are lowered.

Description

technical field [0001] The invention relates to a method for preparing porcine type II circovirus ORF2 protein. Background technique [0002] Porcine circovirus type II (PCV2) is one of the newly discovered animal viruses in recent years. It belongs to the Circoviridae family. , is the smallest animal virus found so far. The disease was first discovered in western Canada in 1991, and is divided into PCV-1 and PCV-2 according to the pathogenicity. Among them, PCV-2 has strong infectivity to pigs, and can infect pigs of different ages through the oral and respiratory routes, causing multisystemic wasting syndrome (PMWS) in weaned piglets. Infected pigs have progressive emaciation, sluggishness, dyspnea, jaundice, pale skin, and even death. Etiological and serological investigations have shown that the disease is widespread in many countries and has caused great losses to the world's pig industry. [0003] The PCV-2 genome contains ORF1 and ORF2. ORF1 encodes the Rep protein...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/34C12N15/70C07K14/01C07K1/22
Inventor 李朝阳李明义石乔
Owner SHANDONG SINDER TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap