Nematocide crystallin gene cry003-148 and application thereof

A crystal protein, nematicide technology, applied in nematicide, application, genetic engineering and other directions, can solve problems such as inability to effectively control agricultural pests, and achieve the effect of wide application value

Active Publication Date: 2013-12-04
HUAZHONG AGRI UNIV
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Problems solved by technology

However, with the continuous promotion of the use of Bt insecticides, a variety of insects have developed resistance. In addition, the existing insecticidal crystal proteins are still unable to effectively control certain agricultural pests. Therefore, more new insecticides are sought and cloned. Insect crystal protein gene has become a key way to prevent and control the resistance of target pests to Bt insecticides, and is the core content of various control strategies

Method used

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  • Nematocide crystallin gene cry003-148 and application thereof
  • Nematocide crystallin gene cry003-148 and application thereof
  • Nematocide crystallin gene cry003-148 and application thereof

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Embodiment 1

[0021] Cloning of insecticidal crystal protein gene cry003-148 in Bacillus thuringiensis Sbt003 in embodiment 1

[0022]The present invention uses wild-type Bacillus thuringiensis Sbt003 (the wild-type Bacillus thuringiensis (Bacillus thuringiensis) Sbt003 was sent to China. Wuhan. Wuhan University China Type Culture Collection Center for preservation on May 15, 2012, and its preservation number is CCTCC NO :M2012168) as a nematocidal crystal protein gene The source strain of this strain can produce spherical, rhombic and long grain-shaped parasporal crystals during spore formation. Bioassay experiments show that its crystal protein has high activity against Caenorhabditis elegans and Meloidogyne incognita.

[0023] 1. Extraction of Total DNA from Bacillus thuringiensis Sbt003

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Abstract

The invention belongs to the technical field of microbe genetic engineering, and discloses separation and cloning of nematicidal crystallin gene originated from thuringiensis and biological activity of the nematicidal crystal protein gene on nematode. In the invention, a new insecticidal crystallin gene cry003-148 is obtained by being separated from thuringiensis Sbt003 (the preservation number in China Center for Type Culture Collection is CCTCC NO: M2012168); an encoded product of the gene is a new insecticidal crystallin Cry003-148; and the crystalline has high insecticidal activity on root-knot nematodes in south of China. The invention also comprises preparation of thuringiensis recombinant bacteria BMB1361 which expresses the insecticidal crystallin Cry003-148, the recombinant bacteria is preserved in China Center for Type Culture Collection (CCTCC) and the preservation number is CCTCC NO: M2012057.

Description

technical field [0001] The invention belongs to the fields of agricultural biological control and agricultural microorganism genetic engineering. It specifically relates to the isolation and cloning of an insecticidal crystal protein gene with nematicide activity and the application of the encoded protein in microbial insecticides and transgenic crops. Background technique [0002] Bacillus thuringiensis (Bt for short) is a kind of Gram-positive bacteria widely distributed in the environment. It can form endogenous spores in the middle and late stages of its growth and development, and can produce a type of spore along with the formation of spores. Insecticidal Crystal Proteins (ICPs) that have deliberate poisonous activity against various insects are different from Bacillus cereus (Bc) and Bacillus anthracis (Ba). Bt was first isolated from silkworm larvae by Japanese scholars in 1901, and was officially named Bacillus thuringiensis in 1915 (Yu Ziniu, 1990). Currently, mor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/325C12N15/32C12N1/21C12N1/15C12N1/19A01H5/00A01N63/02A01P5/00C12R1/07
Inventor 孙明刘盈盈叶伟星彭东海石建伟阮丽芳
Owner HUAZHONG AGRI UNIV
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