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Bacillus thuringiensis nematocide crystal protein gene cry1518-35 and uses thereof

A technology of crystal protein and bacillus, applied in the direction of nematocide, application, genetic engineering, etc., can solve unclear problems and achieve the effect of wide application value

Inactive Publication Date: 2009-07-29
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In further gene cloning, two crystal protein genes cry5B and cry6Aa2 with nematicidal activity were found in this strain, but whether there are other genes with insecticidal activity in this strain is not clear from previous studies

Method used

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  • Bacillus thuringiensis nematocide crystal protein gene cry1518-35 and uses thereof
  • Bacillus thuringiensis nematocide crystal protein gene cry1518-35 and uses thereof
  • Bacillus thuringiensis nematocide crystal protein gene cry1518-35 and uses thereof

Examples

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Embodiment 1

[0021] Example 1 Cloning of nematocidal crystal protein gene cry1518-35 in Bacillus thuringiensis YBT-1518

[0022] The present invention uses Bacillus thuringiensis YBT-1518 as the source strain of the nematode crystal protein gene cry1518-35, and the source of the strain can be found in the literature: Yu, Z., P. Bai, W. Ye, F. Zhang, L. Ruan, Z. Yu, and M. Sun. A Novel Negative Regulatory Factor for Nematicidal Cry Protein Gene Expression in Bacillus thuringiensis. J. Microbiol. Biotechnol. 18(6): 1033-1039. The strain has no flagella, can form typical long rice-like crystals, and is highly virulent to root-knot nematodes and Caenorhabditis elegans.

[0023] 1. Extraction of the total plasmid of Bacillus thuringiensis YBT-1518

[0024] Bacillus thuringiensis strain YBT-1518 was activated overnight, and transferred to 5 mL of fresh LB medium at an inoculum size of 1 / 100 (v / v) (LB medium formula: tryptone 1%, yeast extract 0.5%, Sodium chloride 0.5%, pH 7.0), 28 ℃, 200rpm c...

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Abstract

The invention belongs to the technical field of microorganism genetic engineering and discloses separation and cloning of nematicidal crystal protein gene originated from thuringiensis and biological activity of the nematicidal crystal protein gene on nematode. In the invention, a new insecticidal crystal protein gene cry1518-35 is obtained by being separated from bacterial strain of activated thuringiensis YBT-1518 of root-knot nematodes in north of China; an encoded product of the gene is a new insecticidal crystal protein Cry1518-35; the protein Cry1518-35 features high insecticidal activity on root-knot nematodes in north of China. The invention also includes the preparation of colon bacillus recombinant bacteria EMB0225 which expresses the insecticidal crystal protein Cry1518-35, the recombinant colon bacillus is preserved in CCTCC, the preservation number is CCTCC NO: M209009.

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 its encoded protein in microbial insecticides. Background technique [0002] Bacillus thuringiensis (Bt) is a kind of Gram-positive bacteria that widely exists in soil. During its growth and development, it can form endophytic spores, and it is accompanied by the formation of spores to produce specific toxicity to insects. Insecticidal Crystal Proteins (ICPs) are different from Bacillus cereus which secretes enterotoxin and Bacillus anthracis which causes anthrax. Studies have shown that the crystal protein produced by Bacillus thuringiensis can treat more than 500 kinds of insects in 10 orders of Insecta, such as Lepidoptera, Diptera, Coleoptera, Hymenoptera, and Homoptera, as well as protoz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/32C12N1/21C12N1/15C12N1/19A01H5/00C07K14/325A01N63/02A01P5/00C12R1/19
CPCA01N63/02C07K14/325A01N63/50A01N63/23
Inventor 孙明郭素霞彭东海王鹏霞阮丽芳喻子牛
Owner HUAZHONG AGRI UNIV
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