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Method for cloning aldo-keto reductase gene of unknown bacterium

A technique of reducing enzymes and bacteria, applied in the field of molecular biology, to achieve the effect of enriching candidate gene resources

Inactive Publication Date: 2019-02-26
HUBEI UNIV OF TECH
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Problems solved by technology

At present, the method of cloning the Aldoketone reductase gene is based on searching for the Aldoketone reductase gene sequence of a specific bacterium in the GeneBank database, and then designing primers to obtain the Aldoketone reductase gene through PCR amplification. The premise of this method It is necessary to determine the cloning of the aldoketone reductase gene. For unknown strains, this method cannot be used to clone the aldoketone reductase gene

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  • Method for cloning aldo-keto reductase gene of unknown bacterium
  • Method for cloning aldo-keto reductase gene of unknown bacterium
  • Method for cloning aldo-keto reductase gene of unknown bacterium

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[0018] In order to better understand the present invention, the invention will be described in detail below in conjunction with the accompanying drawings and specific examples.

[0019] In order to solve the problem of being unable to clone the aldoketone reductase gene of unknown bacteria, the present invention provides a method for cloning the aldoketone reductase gene of unknown bacteria. The conserved sequence of the Aldoketone reductase gene, the first degenerate primer was designed by saving the sequence, and the part of the Aldoketone reductase gene of the unknown bacterium was cloned by PCR technology, and the bacterial species closest to the unknown bacterium was determined by BLAST comparison , and then design the second degenerate primer according to the Alduoketone reductase gene of the closest bacterial species, and then clone the Alduoketone reductase gene of the unknown bacterium. The following specific examples will be used to clone the unknown bacterium of the ...

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Abstract

The invention relates to the field of molecular biology, in particular to a method for cloning an aldo-keto reductase gene of an unknown bacterium. The method for cloning the aldo-keto reductase geneof the unknown bacterium comprises the following steps: 1), amplifying by a PCR technology, wherein a primer adopts a first degenerate primer and a template is a genomic DNA of the unknown bacterium;2), performing BLAST comparison on a part of the amplified aldo-keto reductase gene to obtain a culture with the closest genetic relationship and the sequence of the intact aldo-keto reductase gene ofthe culture with the closest genetic relationship; 3), according to the sequence of the intact aldo-keto reductase gene of the culture with the closest genetic relationship in the step 2), designinga second degenerate primer, and amplifying the intact aldo-keto reductase gene of the unknown bacterium by the PCR technology, wherein in the PCR amplification technology in the step, the primer is the second degenerate primer and a template is a genomic DNA of the unknown bacterium.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a method for cloning the aldoketone reductase gene of unknown bacteria. Background technique [0002] Aldoketone reductase gene sequences exist in the vast majority of bacteria, and there are differences in the gene sequences of aldoketone reductase in different bacterial species. At present, the method of cloning the Aldoketone reductase gene is based on searching for the Aldoketone reductase gene sequence of a specific bacterium in the GeneBank database, and then designing primers to obtain the Aldoketone reductase gene through PCR amplification. The premise of this method It is necessary to determine the strain of cloning the aldoketone reductase gene. For unknown strains, this method cannot be used to clone the aldoketone reductase gene. Contents of the invention [0003] In order to solve the above problems, the object of the present invention is to provide a method for c...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/10
CPCC12Q1/686C12Q2565/125
Inventor 宫春杰王崇鞠
Owner HUBEI UNIV OF TECH
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