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Method for rapid amplifying nemato phagous fungi extracellular serine proteinase gene and its use

A technology of serine protease and nematophagous fungi, which is applied in the fields of molecular biology and applied microbiology, and can solve the problems of undiscovered public reports and the like

Inactive Publication Date: 2005-12-21
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Through document search, do not find the public report of the same document as the content of the present invention

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Amplification of the Conserved Sequence of the Extracellular Serine Protease Gene of Verticillium ciferium

[0047]Inoculate the bacterium of Verticillium ciferium (L.psalliotae, syn.V.psalliotae) on a PDA plate, culture at 26°C for 6-8 days, collect the hyphae and extract genomic DNA, and use the genomic DNA of Verticillium cannibalum As a template, primer 1 was used to amplify to obtain the conserved sequence of the gene encoding extracellular serine protease.

Embodiment 2

[0048] Example 2: Amplification of the Conserved Sequence of the Extracellular Serine Protease Gene of Monocremonium cystospora

[0049] Inoculate M.cystosporium cells on a PDA plate, culture at 26°C for 6-8 days, collect mycelium and extract genomic DNA, and use primer 2 to amplify to obtain the conservation of the extracellular serine protease coding gene sequence.

Embodiment 3

[0050] Example 3: Amplification of the extracellular serine protease gene of Separation sp. YMF1.00118

[0051] Basically the same as in Example 2, using the genomic DNA of Dactylella spp. YMF1.00118 as a template, amplified with primer 3 to obtain the full sequence of the extracellular serine protease mature peptide and a partial sequence of the propeptide.

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PUM

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Abstract

The invention relates to the method and application of fast expandation and increasing outside of the food line insect fungus butcher serine the protein enzyme gene, which belongs to the molecular biology and the application of microbiology domain. This invention design 5 pair of degenerations directs outside of the butcher which the thing (altogether 6) were used in fast expandation and increasing and preys on the line insect fungus serine protein enzyme gene. This invention design 5 pair of degenerations direct sequence is: Directs thing 1, AARTAYATYGTCAAGYWSAAG / AYRTGDGGRSWRGCCATSGA; Directs thing 2, AARTAYATYGTCGTCYWSAAG / AYRTGDGGRSWHGCCATSGA; Directs thing 3, AARTAYATYGTCGTCYWSAAG / TTAAGYRKHKCCRTTGWAG; Directs thing 4, MAWGMTKWYGAACSKYCT / GTRTCRATGACGTAGACKG; Directs thing 5, MAWGMTKWYGAACSKYCT / TTAAGYRKHKCCRTTGWAG.

Description

Technical field: [0001] The invention relates to a method and application for rapidly amplifying the extracellular serine protease gene of nematode fungus, belonging to the fields of molecular biology and applied microbiology. Background technique: [0002] Plant parasitic nematodes are a large class of important plant diseases, causing huge losses every year. In the biological control of nematodes, nematode fungi are a very important type of nematode biocontrol resources. At present, commercial nematode biocontrol agents, including nematode Bike, have been put on the market. [0003] During the nematode infestation process, the body wall or egg shell covering the surface is an important protective structure to prevent infestation, no matter for the nematode body or eggs. The body wall of nematodes is mainly composed of proteins and fibers including keratin and collagen, and the egg shell is mainly composed of chitin layers. The body wall and egg shell together form the ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/57
Inventor 张克勤杨金奎
Owner YUNNAN UNIV