Soluble expression and purification method for recombinant murine leukemia virus reverse transcriptase (MMLV-RT) in colon bacillus

A murine leukemia virus, MMLV-RT technology, applied in the field of biochemistry, can solve the problems of easy formation of inclusion bodies, low yield, complex natural reverse transcriptase extraction technology, etc., and achieve the effect of increased solubility and simplified purification
CN101684474AInactive Publication Date: 2010-03-31孙启明 +1
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
孙启明
Publication Date
2010-03-31
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to an expression and purification of murine leukemia virus reverse transcriptase (MMLV-RT), in particular to a soluble expression and a purification method for recombinant murineleukemia virus reverse transcriptase (MMLV-RT), which belongs to the field of biochemistry. MMLV-RT genes and enterokinase restriction enzyme cutting sites are cloned in a pET28a vector through vector construction; the MMLV-RT and molecular chaperones are co-expressed at the low temperature; the expression amount of protein is improved to 15 percent; and the solubility of the protein is 10 timesthat of the protein expressed by a conventional method.
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Description

technical field

[0001] The invention relates to the expression and purification of murine leukemia virus reverse transcriptase (MMLV-RT), in particular to a soluble expression and purification method of murine leukemia virus reverse transcriptase (MMLV-RT). Belongs to the field of biochemistry. Background technique

[0002] As an important molecular biology research method, RT-PCR is a technology that combines reverse transcription (RT) of RNA and polymerase chain amplification (PCR) of cDNA. First, cDNA is synthesized from RNA by the action of reverse transcriptase, and then cDNA is used as a template to amplify and synthesize the target fragment. RT-PCR technology is sensitive and versatile, and can be used to detect the level of gene expression in cells, the content of RNA viruses in cells and directly clone the cDNA sequence of a specific gene. The process from mRNA to cDNA is called reverse transcription, which is catalyzed by reverse transcriptase. Reverse transcrip...

Claims

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