Method for inducing silent gene expression of marine actinomycetes to generate new substances

Abstract

The invention belongs to the technical field of gene induction, and discloses a method for inducing a silent gene expression of marine actinomycetes to generate new substances. The method comprises the following steps: flushing a anthocidaris crassispina with sterile aged seawater, washing the anthocidaris crassispina with sterile water for 3 times, then placing the intestinal tissue thereof in the sterile water, diluting, then sucking 200 [mu]L and coating on a Gorodkowa medium plate containing 50 g / mL of potassium dichromate and 20 g / mL of kanamycin to obtain actinomycete strains; performinga small-scale fermentation on the strains, screening the ethyl acetate crude extract of fermentation liquids of 2 strains by adopting a plate filter paper method, and performing an activity screening; and detecting a new secondary metabolite generated by the induced strains. According to the method, a physical and chemical method is firstly adopted to induce the silent gene expression of marine actinomycetes to generate new substances, and a method for inducing and identifying the new substances generated by the marine actinomycetes is established through the combination of a spectral chemistry testing and an activity evaluation.

Description

technical field [0001] The invention belongs to the technical field of gene induction, and in particular relates to a method for inducing marine actinomycetes to express silent genes and produce new substances. Background technique [0002] At present, the existing technology commonly used in the industry is as follows: more than 50% of the active substances produced by microorganisms come from actinomycetes. Due to the outstanding ability of actinomycetes in producing new active metabolites, scientists have We have been working on the isolation of novel actinomycetes from terrestrial microbial sources. However, the number of novel compounds isolated from terrestrial actinomycetes has continued to decrease in recent years, so there is a need to find novel actinomycetes from unexplored actinomycete habitats as a source for the development of novel active secondary metabolites . Due to the particularity of the marine environment, marine actinomycetes differ from terrestrial ...

AI Technical Summary

Problems solved by technology

[0007] (1) The yield of active ingredients produced by fermentation of wild actinomycete strains is often low, and sometimes even new compounds can be isolated, but the amount is too small to complete the work on their activity identification and other aspects

[0008] (2) Many secondary metabolic pathways in actinomycetes are silent under conventional culture conditions, so it is difficult to discover their ability to produce new active substances, thus limiting the discovery of original new drug lead compounds at the source

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