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Method for improving prokaryotic expression of antiviral activity of porcine interferon

A technology of antiviral activity and porcine interferon, applied in the direction of interferon, antiviral agent, microorganism-based method, etc.

Inactive Publication Date: 2012-01-25
SHANGHAI JIAO TONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in practical applications, it is often only a single application of a certain type of interferon

Method used

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  • Method for improving prokaryotic expression of antiviral activity of porcine interferon
  • Method for improving prokaryotic expression of antiviral activity of porcine interferon
  • Method for improving prokaryotic expression of antiviral activity of porcine interferon

Examples

Experimental program
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Embodiment 1

[0033] 1. Construction of prokaryotic recombinant expression plasmids of porcine IFN-α and IFN-γ genes

[0034] Extract porcine leucocyte DNA and porcine spleen RNA, and amplify porcine IFN-α and IFN-γ genes with specific primers. The results are as follows: figure 1shown. The IFN-α and IFN-γ genes were inserted into the prokaryotic expression vector pET-30 to construct the recombinant expression plasmids pET-30-IFN-α and pET-30-IFN-γ, which were transformed into Escherichia coli BL21 to express porcine IFN under the induction of IPTG -α and IFN-γ, SDS-PAGE analysis of recombinant expressed proteins. The result is as figure 2 shown.

[0035] 2. Induced expression of recombinant plasmids

[0036] Take 100 μl of the recombinant expression plasmid bacterial liquid with correct sequencing results and add it to 5 ml kanamycin-resistant LB culture medium to shake the bacteria overnight, and then insert it into 200 ml kanamycin-containing LB culture medium at a ratio of 1:10037 ...

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Abstract

The invention discloses a method for improving the prokaryotic expression of the antiviral activity of a porcine interferon, and the method comprises the following steps: extracting DNA (deoxyribonucleic acid) of porcine white cells and RNA (ribonucleic acid) of porcine spleen, constructing a recombinant expression plasmid, transforming Escherichia coli and inducing the expression of porcine IFN (immunoreactive fibronectin)-alpha and IFN-gamma; and when the weight ratio of the recombinant porcine IFN-alpha to the IFN-gamma is 53:1, and the activity of inhibiting the replication of astrovirus is best. The scheme of the invention provides a production and preparation method of the porcine interferon which is high in expression level and great in purity and a using method for improving the antiviral activity of the interferon, thereby laying a foundation for development of novel gene engineering antiviral preparations, as well as prevention and treatment of porcine infectious diseases.

Description

technical field [0001] The invention relates to a method for improving the antiviral activity of prokaryotically expressed porcine interferon, in particular to a method for improving the antiviral activity of prokaryotically expressed porcine IFN-α and IFN-γ, and belongs to the field of biotechnology. Background technique [0002] my country is a large pig raising country, and various diseases of pigs, especially viral diseases, have seriously endangered the development of the pig industry. To this end, it is very important to develop antiviral and immune enhancing drugs with high efficiency and low cost. Interferon is a secreted protein secreted by the animal body under the action of inducing factors, which has various biological activities such as immune regulation function, broad-spectrum anti-virus, and anti-tumor. It is an important part of the body's defense system. Its anti-virus, anti-tumor and anti-parasitic effects have been extensively researched and affirmed. P...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C07K14/56A61P31/14C07K14/57C12R1/19
Inventor 朱建国叶承荣俞向前赵伟鸽林源
Owner SHANGHAI JIAO TONG UNIV
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