Streptomyces antibiotic related regulatory gene screening method

Abstract

The invention discloses a streptomyces antibiotic related regulatory gene screening method. The method comprises the following steps: evaluating the antibiotic synthesis difference between a global regulatory gene and an original strain after the global regulatory gene is knocked out, analyzing and screening genes with abnormal and obvious up-regulation and down-regulation by utilizing multiomics, cloning the screened genes to a high-expression vector pSN7, performing high expression in streptomyces, and detecting the yield of antibiotics by using HPLC (High Performance Liquid Chromatography). The strains with obviously increased yield are recorded, mutation of modification sites is carried out on corresponding genes, and then high-yield strains can be obtained after high expression, yield detection and screening are repeated. The method is different from a method which is only limited to synthesis of gene cluster fishing protein through experiments, can be used for screening genes beneficial to increase of the yield of antibiotics in a wider range, and provides a direction for mutation of the genes in a multi-omics manner. The method can be used for detecting and screening any medicinal microorganisms, is wide in applicability and relatively high in extensibility, and can be used for comparing various knockout strains according to changes of different degrees.

Description

technical field [0001] The invention relates to the fields of biochemistry and molecular biology, in particular to a method for screening Streptomyces antibiotic-related regulatory genes. Background technique [0002] Thousands of genes are transcribed and translated in microorganisms, and they form complex regulatory networks that manage cellular metabolism. In addition, there are a variety of post-translational modifications that occur on functional proteins, such as phosphate sulfonylation, SUMOylation, formylation, acetylation, propionylation, butyrylation, succinylation, malonylation, crotonylation, glutarylation, etc. to adjust protein activity to meet the needs of different growth stages Or respond to changes in the external environment. [0003] Streptomyces belong to the genus Actinomyces and are Gram-positive bacteria. Streptomyces synthesizes a wide variety of secondary metabolites, providing a precious treasure trove of antibiotics for clinical treatment. Its ...

AI Technical Summary

Problems solved by technology

[0005] Taking the daptomycin-producing bacteria Streptomyces minor as an example, the whole genome sequencing shows that its gene size is 7.6Mbp, a total of 7084 genes, including a large number of regulatory genes, various regulatory genes in the growth and metabolism of Streptomyces minor A complex regulatory network has been formed in the process. It can be seen that the antibiotic-regulated genes screened so far can only be the tip of the iceberg. Therefore, various efficient regulatory gene screening, positioning and other research methods are urgently needed.

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