Colibacillus plasmid vector and its application method

Abstract

The present invention relates to plasmid vector capable of being used in gene recombinant technology, and the method of utilizing the plasmid vector in expressing target gene. The plasmid vector of the present invention features that it has colibacillus sigma factor sigma32 distinguished and regulated promoter. The method of utilizing the plasmid vector in expressing target gene includes inserting the expression vector gene into one of the plasmid vector of the present invention, transforming colibacillus host cell with the obtained recombinant plasmid and inducing the expression of the target gene in heat shock inducing method during the culture. The bioactive protein gene expression of the present invention has the advantages of low background expression, high recombinant protein yield, no use of chemical inducing agent, easy molecular operation, and no need of special gene reformation of host strain.

Description

technical field

[0001] The present invention relates to the field of genetic engineering, specifically, to a plasmid vector that can be used in gene recombination technology and its application method, more specifically, to a plasmid vector regulated by the Escherichia coli heat shock system, and to using the plasmid vector to express A method for producing a target protein from an exogenous gene. Background technique

[0002] Escherichia coli expression system consists of two parts: expression vector and corresponding host cells. It is the earliest, most widely used and easiest-to-operate high-efficiency gene expression system. In the E. coli expression system, expression vectors usually use regulatable promoters to control the expression of foreign genes; the types of these promoters are important for controlling the inhibition of cell growth by the expressed target protein and reducing the degradation of the target protein by cellular proteases , and the yield and solubi...

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