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Method for cloning member group of isopentenyl transferase gene family

A gene family and transferase technology, applied in the field of molecular biology, can solve the problems of not being able to fully obtain the members of the isopentenyl transferase gene family, the limited number of prenyl transferase genes, and the inability to obtain the isopentenyl transferase gene. Achieve the effect of enriching candidate gene resources and increasing types

Pending Publication Date: 2020-03-13
HUBEI UNIV OF TECH
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Problems solved by technology

At present, the method for cloning prenyltransferase gene family members is first to search for prenyltransferase gene sequence in the gene bank, and then design primers to clone prenyltransferase gene according to the prenyltransferase gene sequence; this method The obtained prenyl transferase genes are very limited, only limited to the published prenyl transferase genes in the gene bank, and the undisclosed prenyl transferase genes in the gene bank cannot be obtained, and the prenyl transferase genes cannot be fully obtained gene family members

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  • Method for cloning member group of isopentenyl transferase gene family
  • Method for cloning member group of isopentenyl transferase gene family
  • Method for cloning member group of isopentenyl transferase gene family

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Embodiment Construction

[0016] 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.

[0017] A method for cloning a member of the prenyltransferase gene family, comprising the steps of:

[0018] (1) select bacteria one: bacteria Cryobacterium sp.GCJ06;

[0019] (2) Utilize PCR technology to amplify part of the isopentenyl transferase 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;

[0020] 2.1) The design process of the base sequence of the first degenerate primer:

[0021] Using the NCBI (https: / / www.ncbi.nlm.nih.gov / ) database, using "Al / ketoreductase" as the search term, searched for "Nucleotide" and "Protein", and obtained multiple...

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Abstract

The invention relates to the field of molecular biology, in particular to a method for cloning a member group of an isopentenyl transferase gene family. The method for cloning a member group of an isopentenyl transferase gene family comprises the following steps: 1) selecting a bacterium 1; 2) amplifying part of the isopentenyl transferase genes of the bacterium 1 by adopting a first degenerate primer as the primer and the genome of the bacterium 1 as the template; 3) comparing part of the amplified isopentenyl transferase genes to obtain a strain with the closest genetic relationship and obtain a complete isopentenyl transferase gene sequence thereof; 4) designing a second degenerate primer according to the complete isopentenyl transferase gene sequence of the most closely related strain,and amplifying the complete isopentenyltransferase gene of the bacterium 1, wherein the template is the genome of the bacterium 1; and 5) obtaining a complete isopentenyltransferase gene of a plurality of bacteria sequentially according to the method, and constructing the member group of the isopentenyl transferase gene family. The method of the invention can be adopted to obtain a large number of isopentenyl transferase gene family members.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a method for cloning prenyltransferase gene family members. Background technique [0002] Prenyltransferase is an important catalyst for the production of statins. Obtaining the members of the prenyltransferase gene family can provide abundant candidate gene resources for the study of prenyltransferase genes and the production of statins. At present, the method for cloning prenyltransferase gene family members is first to search for prenyltransferase gene sequence in the gene bank, and then design primers to clone prenyltransferase gene according to the prenyltransferase gene sequence; this method The obtained prenyl transferase genes are very limited, only limited to the published prenyl transferase genes in the gene bank, and the undisclosed prenyl transferase genes in the gene bank cannot be obtained, and the prenyl transferase genes cannot be fully obtained gene family membe...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/54
CPCC12N15/11C12N9/1085C12Y205/01
Inventor 宫春杰王崇鞠
Owner HUBEI UNIV OF TECH
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