Stress, signaling and transcriptional regulatory genes involved in regulating act production in Streptomyces

A technology of transcriptional regulation and Streptomyces, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve problems such as uncertainty, further research and discussion, and complexity of gene regulation functions

Active Publication Date: 2022-06-21
HENAN BUSINESS SCI RES INST
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  • Abstract
  • Description
  • 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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  • Stress, signaling and transcriptional regulatory genes involved in regulating act production in Streptomyces

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Experimental program
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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 for the unreported process, please refer to "Using Transposition Mutagenesis to Study the Functional Genome of Streptomyces coelicolor A3(2)" (Xu Zhong, Master Thesis of Huazhong Agricultural University 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 sp. through the conjugation of Escherichia coli strain ET12567 / pUZ8002 and Streptomyces coelicolor, and the ratio of recipient bacterium to donor bacterium was 1:1;

[0063] Theoretically, after entering Streptomyces, the transposase Tnp(5) begins to be expressed, and the mini-Tn5 fragment is sheared and randomly inserted into the genome of Streptomyces. Since the plasmid cannot replicate and integrate in S...

Embodiment 2

[0077] In this embodiment, the relevant screening results of Embodiment 1 are briefly introduced and described as follows.

[0078] On the basis of the screening method in Example 1, 988 strains with variable actiniodopsin production were screened out of 50,000 mutants. The transposition insertion site mapping of these strains showed that these mutant strains involved 551 genes; some of them involved 551 genes. The specific explanations of genes that have regulatory effects on actiniodopsin production are as follows.

[0079] Cells are synthetically regulated by:

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

[0081]

[0082] Note (the data in the following table have the same meaning and will not be explained again): The relative production of ACT in the table represents the ratio of ACT production of the mutant strain to that of the wild-type strain. Less than 1 indicates that the ACT production o...

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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 stress, signal and transcription regulation genes in the regulation of Streptomyces ACT production. There are 14 stress, signal and transcription regulation genes in total; mutations of such genes have regulatory effects on the ACT production of Streptomyces. 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 present application belongs to the technical field of microbial fermentation engineering, and specifically relates to a gene related to the production of polyketide ACT (ACT) in Streptomyces sp. 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-infective and anti-tumor drugs are derived from microbial secondary metabolites. With more and more microbial genome sequencing work, 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...

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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