Method for constructing bacillus licheniformis by knocking out lrpC gene, strain and application of strain

A technology of Bacillus licheniformis and Bacillus licheniformis, applied in the direction of microorganism-based methods, other methods of inserting foreign genetic materials, applications, etc., can solve problems that cannot be deduced and have not yet resolved the regulation mechanism of LrpC

Active Publication Date: 2018-08-24
LIFECOME BIOCHEM
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current research has not resolved the specific regulatory mechanism of LrpC, therefore, it is impossible to deduce the relationship between LrpC and leucine synthesis and bacitracin production

Method used

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  • Method for constructing bacillus licheniformis by knocking out lrpC gene, strain and application of strain
  • Method for constructing bacillus licheniformis by knocking out lrpC gene, strain and application of strain
  • Method for constructing bacillus licheniformis by knocking out lrpC gene, strain and application of strain

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Experimental program
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Embodiment Construction

[0025] by knockout wxya The method for genetically constructing bacillus licheniformis comprises the following steps:

[0026] (1) Using the genomic DNA of Bacillus licheniformis DW2 as a template, PCR amplified wxya The upstream homology arm of the gene and wxya The downstream homology arm of the gene; and then use the overlap extension PCR method to wxya The upstream homology arm of the gene and wxya The downstream homology arms of the gene are spliced ​​together to obtain the fusion gene sequence A;

[0027] (2) Using restriction endonuclease Xba I and Sac I Carrying out double digestion on the fusion gene sequence A obtained in step (1) to obtain the restriction gene sequence A;

[0028] (3) Prepare plasmid T2(2)-ori, and use restriction endonuclease Xba I and Sac I Carry out double digestion of the plasmid T2(2)-ori to obtain the restriction plasmid T2(2)-ori;

[0029] (4) Link the enzyme-cut gene sequence A obtained in step (2) to the enzyme-cut plasmid T2(2...

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Abstract

The invention provides a method for constructing bacillus licheniformis by knocking out an lrpC gene, a strain and an application of the strain. The bacillus licheniformis DW2 delta lrpC is obtained by knocking out the lrpC gene in a bacillus licheniformis genome with a genetic engineering method, and the yield of bacitracin of the strain in a fermentation liquid in the bacitracin fermentation process is increased by 10% or above.

Description

technical field [0001] The invention relates to the field of Bacillus licheniformis strain transformation, especially a wxya The method and strain of gene constructing Bacillus licheniformis and its application. Background technique [0002] Bacillus licheniformis belongs to Gram-positive bacteria, which is recognized as an important industrial microbial strain with biosafety (GRAS). Because of its clear genetic background and high industrial application value, Bacillus licheniformis has been widely used for research in recent years. [0003] At present, Bacillus licheniformis is mainly used to ferment and produce biochemical products such as poly-γ-glutamic acid, bacitracin, acetoin, 2,3-butanediol, and lichenin. Bacitracin, also known as subtilisin, can inhibit or kill certain pathogenic bacteria, strongly inhibit the growth of Gram-negative bacteria, and have synergistic enhancement effects with other antibiotics (such as penicillin, gentamicin, etc.); and , which is h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/75C12N15/90C12N15/66C12N15/31C12N1/21C12P21/04C12R1/10
CPCC07K7/58C07K14/32C12N15/66C12N15/75C12N15/902C12P21/02
Inventor 陈守文蔡冬波朱江李阳朱杉李俊辉楼丽君陈晓斌
Owner LIFECOME BIOCHEM
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