Genetic manipulation method for manually controlling spontaneous mutation rate of bacillus subtilis and application thereof

Abstract

The invention belongs to the technical field of biology, and particularly relates to a genetic manipulation method for manually controlling a spontaneous mutation rate of bacillus subtilis and application thereof. The genetic manipulation method comprises the following steps: regulating and controlling an operon MutSL which is responsible for a mismatch repair system of the bacillus subtilis by axylose induced promoter and repressor protein, and establishing recombinant plasmids in corresponding inducible expression; integrating the recombinant plasmids to a bacterial strain genome by utilizing Spizizen double exchange, and carrying out positive-negative selection, thus obtaining objective engineering strains; regulating different expression levels of the operon MutSL of the mismatch repair system through different xylose addition concentration, thus realizing manual control on the spontaneous mutation rate of the bacillus subtilis.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a genetic manipulation method for artificially controlling the spontaneous mutation rate of Bacillus subtilis and its application. Background technique [0002] Bacillus subtilis ( Bacillus subtilis ) is a dominant microbial species that exists in soil and plant micro-ecosystems. It is non-pathogenic, harmless to humans and animals, does not pollute the environment, and has strong stress resistance and antibacterial and disease-preventing effects. It is widely used as a molecular biology Research model organisms and industrial production of high-yield strain hosts. In recent years, with the widespread application of second-generation and third-generation high-throughput sequencing technologies, studies such as adaptive laboratory evolution have increasingly become research hotspots. Adaptive laboratory evolution is a method of artificially simulating the variation and selection pro...

AI Technical Summary

Problems solved by technology

For adaptive laboratory evolution, gene mutation is the main driving force for its evolution, but in the evolution process, in order to maintain the integrity of its own genome, there is a strict mismatch repair mechanism in the cell to eliminate replication and recombination events Mismatched regions generated in , thereby reducing the natural mutation rate of the cell (-6 ), resulting in a long evolutionary cycle

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