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A transporter-encoding gene involved in the regulation of act production in Streptomyces

A technology for encoding genes and transporters, applied in the field of microbial fermentation engineering, which can solve problems such as uncertainty, the complexity of gene regulation functions, and the need for further in-depth research and discussion.

Active Publication Date: 2021-09-17
HENAN BUSINESS SCI RES INST
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  • Claims
  • Application Information

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Problems solved by technology

[0004] In the prior art, the inventors used transposition mutagenesis technology to study the functional genome of Streptomyces coelicolor A3(2), and initially constructed a mutant library of Streptomyces coelicolor A(3)2 (for details, please refer to : Xu Zhong, using transposition mutagenesis to study the functional genome of Streptomyces coelicolor A3 (2), Master thesis of Huazhong Agricultural University in 2011), but due to the complexity and uncertainty of the regulatory functions of related genes, which specific genes are related to The regulation of secondary metabolites still needs further research and discussion

Method used

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  • A transporter-encoding gene involved in the regulation of act production in Streptomyces

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Embodiment 1

[0060] In this example, the mutagenesis process of Streptomyces coelicolor M145 by using the mini-Tn5 transposition system is briefly introduced as follows. It should be noted that if the process is not described in detail, please refer to "Use of transposition mutagenesis to study the functional genome of Streptomyces coelicolor A3 (2)" (Xu Zhong, Huazhong Agricultural University, master's thesis in 2011).

[0061] (1) Transposition mutagenesis of Streptomyces coelicolor by transposon

[0062] First, referring to the prior art, the transposon plasmid pHL734 was introduced into Streptomyces through conjugative transfer of Escherichia coli strain ET12567 / pUZ8002 and Streptomyces coelicolor, and the ratio of recipient bacteria to donor bacteria was 1:1;

[0063] Theoretically, after entering Streptomyces, the transposase Tnp(5) begins to express, and the mini-Tn5 fragment is cut and randomly inserted into the Streptomyces genome. After the seat occurs, it disappears;

[0064] ...

Embodiment 2

[0077] In this embodiment, a brief introduction to the relevant screening results of Embodiment 1 is as follows.

[0078] On the basis of the screening method in Example 1, 988 strains with changes in actinhodine production were screened out from 50,000 mutants, and the transposition insertion site positioning of these strains showed that these mutant strains involved 551 genes; some of them The specific explanation of the genes that have a regulatory effect on the actin rhodopsin production is as follows.

[0079] The cell is synthesized to regulate the class:

[0080] 7 in total, including genes SCO1525 , SCO2097 , SCO2132 , SCO3150 , SCO4440 , SCO4878 , SCO5174 ;

[0081]

[0082] Note (the meanings of the data in the following tables are the same and will not be explained): The relative production of ACT in the table indicates the ratio of ACT production between the mutant strain and the wild-type strain. Less than 1 indicates that the ACT production of t...

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Abstract

The application belongs to the technical field of microbial fermentation engineering, and specifically relates to genes related to the production of polyketide actin (ACT) in streptomyces. This application specifically relates to the application of the transporter gene in the regulation of Streptomyces ACT production, and there are 6 genes encoding the transporter; including: SCO2254 , SCO2534 , SCO3765 , SCO6160 , SCO2519 , SCO3185 ; After the mutation of this type of gene, it has a regulation effect on the production of Streptomyces ACT. The inventor carried out mutagenesis on Streptomyces coelicolor M145, and screened the strains with changed actin rhodopsin production. The position of the transposon on the genome was located by enzyme digestion, self-ligation and sequencing of the screened positive clones, and the principle of multiple screening to locate the same gene was used to exclude the spontaneously mutated gene, and finally determine the factor affecting the production of actinhodine. Gene.

Description

technical field [0001] The application belongs to the technical field of microbial fermentation engineering, and specifically relates to genes related to the production of polyketide actin (ACT) in streptomyces. Background technique [0002] Existing studies generally believe that small molecular compounds produced by microbial secondary metabolism are of great significance to human health. Statistics show that more than 60% of FDA-approved anti-infection and anti-tumor drugs are derived from microbial secondary metabolites. With the sequencing of more and more microbial genomes, genome sequence analysis shows that microorganisms have great potential to produce various secondary metabolites, but some studies have shown that many microbial secondary metabolites cannot be expressed under laboratory culture conditions. , and most environmental microorganisms cannot be effectively cultured under laboratory conditions, thus seriously hindering the development of microbial second...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/31
CPCC07K14/36
Inventor 徐钟李远远王永王法云
Owner HENAN BUSINESS SCI RES INST
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