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Trichoderma chitinase and gene for expressing it

A technology of chitinase and gene, applied in genetic engineering, plant genetic improvement, hydrolytic enzyme, etc.

Inactive Publication Date: 2002-09-18
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The discovery and utilization of the antifungal properties of Trichoderma has a history of nearly 70 years, but the research on the cloning of genes encoding its cell wall degrading enzymes and the application of related genes in antifungal plant genetic engineering has just started, especially in our country, no research reports have been seen

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  • Trichoderma chitinase and gene for expressing it
  • Trichoderma chitinase and gene for expressing it
  • Trichoderma chitinase and gene for expressing it

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Embodiment 1

[0015] Embodiment 1, the construction of the Escherichia coli expression vector of Trichoderma chitinase cDNA

[0016] The plant expression vector pART27 used in the construction of the Trichoderma chitinase gene Tch42 cDNA is used in conjunction with the expression intermediate vector pART7. The pART7 plasmid contains the CaMV 35S promoter and OCS 3' terminator. First, the 1.3 kb Trichoderma chitinase cDNA (Tch42) was excised from the cloning vector pMD18-TTch with EcoRI / BglII, and inserted into the Escherichia coli expression vector pBV221 digested with EcoRI / BamHI (BglII and BamHI are homologous enzymes) The multiple cloning site between the promotor and the terminator, promptly obtains Escherichia coli expression vector pBVTch ( figure 1 ). Embodiment 2, the construction of Trichoderma chitinase cDNA plant expression vector

Embodiment 2

Embodiment 3

[0017] The plant expression vector pART27 used in the construction of the Trichoderma chitinase cDNA is used in conjunction with the expression intermediate vector pART7. The pART7 plasmid contains the CaMV 35S promoter and OCS 3' terminator. First, the 1.3 kb Trichoderma chitinase cDNA (Tch42) was excised from the cloning vector pMD18-TTch with BglII / ClaI, and inserted into the promoter of the intermediate vector pART7 digested by BamHI / ClaI (BglII and BamHI are homologous enzymes) The multiple cloning site between the promoter and the terminator was used to obtain the expression intermediate vector pART7Tch; then the exogenous gene with the promoter and terminator was excised with NotI, and the corresponding NotI single cloning site in the T-DNA region of the pART27 plasmid was used Inserted into the pART27 expression vector to construct the monovalent plant expression vector pART27Tch( figure 2 ). Example 3. Cotton was genetically transformed by laser micro-beam puncture...

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Abstract

A chitinase with higher anti-fungus activity, broad-spectrum resistance and no toxic damage to plant and the gene for expressing it care disclosed. The said gene comes from Trichoderma viride. Through separation, complete gene (42 KD) and cDNA are obtained, and used to congifure colibacillus expression carrier and plant expression carrier, and then obtain anti-fungus transgenic plant.

Description

technical field [0001] The invention relates to a trichoderma chitinase, a gene expressing the enzyme and a method for using the gene to breed transgenic plants resistant to verticillium wilt and fusarium wilt. technical background [0002] Some progress has been made in obtaining antifungal transgenic plants by using chitinase gene and glucanase gene. However, the exogenous genes currently used are mainly derived from plants or bacteria, and the level and range of resistance obtained can rarely meet the requirements of commercial agricultural production. The chitinase and other fungal cell wall degrading enzymes produced by the biocontrol fungus Trichoderma have the advantages of higher antifungal activity, wider resistance spectrum, and no toxicity to plants at high concentrations. The chitinase gene can produce high-level broad-spectrum resistance, which overcomes the limitations of chitinase genes derived from plants and bacteria. transgenic plants. [0003] The disco...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/06A01H5/00C12N9/24C12N15/56C12N15/82
Inventor 陈正华张海燕刘桂珍欧阳藩张铁汉
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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