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Subtilisin-like protease gene recombination disinsection engineering bacteria and method for preparing inocula

A Bacillus subtilis, gene recombination technology, applied in genetic engineering, botanical equipment and methods, microorganism-based methods, etc., can solve the problems of short residual period, weaker virulence than chemical pesticides, and narrow insecticidal spectrum. Achieve the effect of increasing insecticidal virulence, improving biosafety, and expanding the scope of insecticidal spectrum

Inactive Publication Date: 2008-07-30
HUNAN NORMAL UNIVERSITY
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Problems solved by technology

[0006] The purpose of the present invention is to overcome defects such as narrower insecticidal spectrum of existing Bt insecticides, weaker toxicity than chemical insecticides, short residual effect period, etc. Longer subtilisin-like protease gene recombination insecticidal engineering bacteria and its preparation method

Method used

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  • Subtilisin-like protease gene recombination disinsection engineering bacteria and method for preparing inocula
  • Subtilisin-like protease gene recombination disinsection engineering bacteria and method for preparing inocula
  • Subtilisin-like protease gene recombination disinsection engineering bacteria and method for preparing inocula

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Embodiment Construction

[0024] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.

[0025] The subtilisin-like gene recombination insecticidal engineering bacterium in this example is a double-effect engineering bacterium constructed by fusing the cry1Ac gene of the Bacillus thuringiensis 4.0718 strain and the exogenous toxin gene Beauveria bassiana subtilisin-like CDEP2 gene.

[0026] The Beauveria bassiana-like subtilisin-like CDEP2 gene is a new subtilisin-like protease gene with independent intellectual property rights discovered by the inventor in Beauveria bassiana, and its registration on GenBank (GenBank) The number is EF195164.

[0027] In this example, Bacillus thuringiensis highly virulent insecticidal strain 4.0718 (preserved by China Center for Type Culture Collection, preservation number CCTCC No.M200016, see Chinese Patent No. ZL01114592.7) and Bt crystal-free mutant strain Cry-B were used as host bacteria , t...

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Abstract

The invention provides a subtilisin-like gene recombination insect killing engineering bacteria and the microbial inoculums preparing method thereof. The insect killing engineering bacteria is a double-effect engineering bacteria which is fused and constructed by the crylAc gene of bacillus thuringiensis and a foreign toxin gene, and expresses fused insecticidal protein crystals in the bacillus thuringiensis. The bacillus thuringiensis is the 4.0718 strain of bacillus thuringiensis. The foreign toxin gene is the CDEP2 gene of subtilisin-like of spherical spore white muscardine fungi. The titers of subtilisin-like gene recombination insect killing engineering bacteria microbial inoculums prepared by fermenting production are all above 6000 international units / microgram, the insect killing rates on the lepidoptera, dipteral and coleopteran insects all reach 90 percent. Under same consistence, the insect killing effect of the invention is 3-5 times higher than that of the monovalent microbial inoculums. The invention has no toxic residue, is safe to human being and animal, and is conducive to protecting entironment.

Description

technical field [0001] The invention relates to a subtilisin-like protease gene recombined insecticidal engineering bacterium and a preparation method thereof, in particular to a Bacillus thuringiensis engineering bacterium containing exogenous subtilisin-like protease gene recombination and a preparation method for the bacterial preparation thereof. Background technique [0002] Human beings have used a large amount of highly toxic and high-residue chemical pesticides for a long time, and these chemical pesticides have brought well-known negative effects and impacts on the ecological environment, food safety and human health. Therefore, it is imperative to reduce the use of highly toxic and high-residue chemical pesticides. Biological control-based pest control strategies are receiving widespread attention, and the development of efficient and stable microbial pesticides has become a top priority. Microbial pesticides have the characteristics of no pollution, no residue, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/62C12N15/57C12N15/75C12R1/07
Inventor 夏立秋丁学知曾智孙运军张友明
Owner HUNAN NORMAL UNIVERSITY
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