Application of hydrolytic enzyme CwlC in metrocyte lysis of bacillus

A technology of Bacillus cereus and Bacillus cereus, applied in the dissolution of hydrolase, bacteria and microorganisms, etc., can solve the problem of unknown key hydrolase

Active Publication Date: 2017-06-09
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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However, the key hydrolases involved in mothe...

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  • Application of hydrolytic enzyme CwlC in metrocyte lysis of bacillus
  • Application of hydrolytic enzyme CwlC in metrocyte lysis of bacillus
  • Application of hydrolytic enzyme CwlC in metrocyte lysis of bacillus

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[0045] The present invention will be further described in detail below in conjunction with the examples.

[0046] Materials and methods:

[0047] Bacterial strains, plasmids and culture conditions: The bacterial strains and plasmids used in this study are listed in Table 1. Escherichia coli (Escherichia coli, referred to as E.coli) TG1 and BL21 were used as host strains for molecular cloning and protein expression, respectively, and E.coli ET12567 was used for demethylation of plasmid DNA (Hoffmann F, Schmidt M, Rinas U.2000 .Simple technique for simultaneous on-line estimation of biomass and acetate from base consumption and conductivity measurements in high-cell density cultures of Escherichia coli.Biotechnol Bioeng70:358–361.,Wang G,Zhang J,Song F,Wu J,Feng S,Huang D. 2006. Engineered Bacillus thuringiensis GO33A with broad insecticidal activity against lepidopteran and coleopteran pests. Appl Microbiol Biotechnol 72:924–930.). All strains of E.coli were cultured in Luria...

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Abstract

The invention relates to 'application of a hydrolytic enzyme CwlC in metrocyte lysis of bacillus'. The invention identifies a novel cwlC gene which joins in metrocyte lysis in B.thuringiensis, cwlC gene deletion can completely block metrocyte lysis, and the yield of spore formation and Cry1Ac protein are not affected. Meanwhile, a CwlC protein marked by a GFP (Green Fluorescent Protein) can be tightly combined with cell walls. More importantly, walls of cells of B.thuringiensis and Bacillus cereus with activities in logarithmic phases can be hydrolyzed by CwlC. Tests show that CwlC is not only a key point of metrocyte lysis of B.thuringiensis in a spore period and has great significances in establishing B.thuringiensis engineering bacteria for efficient agricultural production, but also has great application prospects in biological medicines and food production.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of hydrolase CwlC in lysing bacillus mother cells. Background technique [0002] Bacillus spp. is a spore-forming Gram-positive bacterium that produces dormant spores in the mother cell when stressed by the external environment. In the late spore stage, mother cell autolysis accompanied by the release of mature spores is a sign of programmed cell death (PCD) (LewisK.2000.Programmed death in bacteria.Microbiol Mol Biol Rev64:503–514 .). Therefore, it is very important to understand PCD controlled by signal transduction pathways in mother cells, and cell wall hydrolases play a key role in this process (Lewis K.2000. Programmed death in bacteria. Microbiol Mol Biol Rev64:503–514 .). Bacterial peptidoglycan hydrolases comprise a large and highly diverse group of enzymes that cleave chemical bonds in high-molecular peptidoglycan or soluble fragments (Shockman GD, Daneo-...

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IPC IPC(8): C12N1/06C12N1/21C12R1/085C12R1/07
CPCC12N1/063C12N9/24
Inventor 宋福平彭琦高坦坦陈晓敏耿丽丽张杰
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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