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Method for cloning farnesyl transferase gene family member group

A technology of farnesyltransferase and gene family, applied in the field of molecular biology, can solve the problem of limited farnesyltransferase gene, inability to fully obtain farnesyltransferase gene family members, and inability to obtain farnesyltransferase gene and other issues to achieve the effect of increasing types and enriching candidate gene resources

Pending Publication Date: 2020-02-21
HUBEI UNIV OF TECH
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  • Claims
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Problems solved by technology

At present, the method of cloning the farnesyltransferase gene family members is to first determine the farnesyltransferase gene sequence in the gene bank, and then design primers to clone the farnesyltransferase of known strains according to the farnesyltransferase gene sequence Gene; the farnesyltransferase gene obtained by this method is very limited, only limited to the disclosed farnesyltransferase gene in the gene bank, and cannot obtain the undisclosed farnesyltransferase gene in the gene bank. Not comprehensive access to farnesyltransferase gene family members

Method used

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  • Method for cloning farnesyl transferase gene family member group
  • Method for cloning farnesyl transferase gene family member group
  • Method for cloning farnesyl transferase gene family member group

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Embodiment Construction

[0015] 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.

[0016] A method for cloning a member of the farnesyltransferase gene family, comprising the steps of:

[0017] (1) select bacteria one: bacteria Cryobacterium sp.GCJ06;

[0018] (2) Utilize PCR technology to amplify the part farnesyltransferase gene of Cryobacterium sp.GCJ06 bacterium, primer adopts the first degenerate primer in the PCR amplification technology, the upstream primer of the first degenerate primer is shown in SEQ ID NO.1 Shown, the downstream primer is shown in SEQ ID NO.2, and the template is the genomic DNA of Bacteria 1;

[0019] 2.1) The design process of the base sequence of the first degenerate primer:

[0020] Using the NCBI (https: / / www.ncbi.nlm.nih.gov / ) database, using "farnesyltransferase" as the search term, searched for "Nucleotide" and "Protein", and obtained multiple f...

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Abstract

The invention relates to the field of molecular biology and particularly relates to a method for cloning a farnesyl transferase gene family member group. The method comprises the following steps: (1)selecting a bacterium 1; (2) amplifying partial farnesyl transferase gene primer of the bacterium 1 by adopting a first degenerate primer as a primer and a genome of the bacterium 1 as a template; (3)comparing the partial farnesyl transferase obtained in amplification to obtain strains having the closest genetic relationship and gene sequences of complete farnesyl transferase of the strains; (4)designing a second degenerate primer according to the gene sequences of the complete farnesyl transferase of the strains having the closest genetic relationship, amplifying the complete farnesyl transferase gene of the bacterium 1 by adopting the genome of the bacterium 1 as the template; and (5) constructing a farnesyl transferase gene family member group by complete farnesyl transferase genes ofa plurality of bacteria according to the method. The method can be used for acquiring a great amount of farnesyl transferase gene family members.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a method for cloning farnesyl transferase gene family members. Background technique [0002] Farnesyltransferase is an important catalyst for the production of statins. Obtaining members of the farnesyltransferase gene family can provide abundant candidate gene resources for farnesyltransferase gene research and production of statins. At present, the method of cloning the farnesyltransferase gene family members is to first determine the farnesyltransferase gene sequence in the gene bank, and then design primers to clone the farnesyltransferase of known strains according to the farnesyltransferase gene sequence Gene; the farnesyltransferase gene obtained by this method is very limited, only limited to the disclosed farnesyltransferase gene in the gene bank, and cannot obtain the undisclosed farnesyltransferase gene in the gene bank. Comprehensive access to farnesyltransferase gen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N15/10C12N15/11
CPCC12N9/1085C12Q1/686C12Q2565/125
Inventor 宫春杰王崇鞠
Owner HUBEI UNIV OF TECH
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