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Anti-aphides gene PPA, preparation thereof and protein encoded thereby

A kind of anti-aphid, gene technology, applied in the direction of genetic engineering, plant gene improvement, application, etc., can solve the problems of incomplete inhibition of development and reproduction, unsatisfactory resistance of aphids and other Homopteran insects, etc.

Inactive Publication Date: 2011-01-19
河北省农林科学院经济作物研究所
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Problems solved by technology

[0005] So far, although many lectin genes with insect resistance activity have been discovered, the resistance to aphids and other Homopteran insects is still not satisfactory
Although lectins have an inhibitory effect on homopteran pests such as aphids, the existing lectins can only inhibit the development of pests to a certain extent, but cannot completely inhibit their development and reproduction. Therefore, how to further improve the anti-insect activity of lectins has become a serious fact before people

Method used

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  • Anti-aphides gene PPA, preparation thereof and protein encoded thereby
  • Anti-aphides gene PPA, preparation thereof and protein encoded thereby
  • Anti-aphides gene PPA, preparation thereof and protein encoded thereby

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Embodiment Construction

[0083] Embodiments of the present invention will be further described below in conjunction with the accompanying drawings, but they are not intended to limit the present invention.

[0084] The anti-aphid gene sequence and encoded protein sequence in this example are as described above, and its specific preparation and anti-aphid function identification include:

[0085] A, 3' RACE amplification of PPA gene;

[0086] B, PPA gene 5' RACE amplification;

[0087] C. Amplification and cloning of the full sequence of the PPA gene.

[0088] Step A is to select the 3'RACE kit provided by TaKaRa Company and follow its instructions; the conventional RNA extraction method extracts the total RNA of the leaves and tubers of Pinellia palmatum, and mixes them in equal amounts for subsequent cDNA synthesis; the specific method As follows: According to the sequence alignment and analysis of the monocotyledon lectin gene sequences of Araceae plants on the International Gene Bank (GenBank), t...

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Abstract

The invention belongs to the preparation of a novel gene, and more particularly relates to an aphid-resistant gene (PPA) and a preparation thereof and a protein using the aphid-resistant gene for coding. The 5' end of TCTAGAATGGCCTCCAAGCTCCTCCTC and the 3' end of CCCGGGC TACGCGGCAATTGGGCGCTT are taken as specific primers, a pinellia pedatisecta schott cDNA is taken as a template and a PCR amplification method is adopted so as to obtain the aphid-resistant gene (PPA); and the aphid-deadly protein is coded by the aphid-resistant gene (PPA). The invention solves the problem that the existing agglutinin gene with insect-resistant activity can not thoroughly inhibit the development and reproduction of insects, has aphid-deadly effect, obtains a full-length PPA gene and a plant expressed carrier constructed therefrom and lays foundation for further transforming the full-length PPA gene and the plant expressed carrier constructed therefrom into important crops such as cotton, wheat and rice to culture homoptera-insect-resistant transgenic plants; and can greatly reduce the use level of agricultural chemicals, achieve the aim of environmental protection and have great economic benefit andapplication value.

Description

technical field [0001] The invention belongs to the preparation of new genes, in particular to the anti-aphid gene PPA, its preparation and the protein coded by it. Background technique [0002] Insect pests are one of the important reasons for the reduction of agricultural production. The global annual cost of managing and using chemical pesticides is as high as 10 billion U.S. dollars. Even so, the losses caused by pests account for 20%-30% of the total crop output every year. Compared with chemical agents, the use of insect-resistant gene resources to carry out crop insect-resistant genetic breeding is the most economical, effective and safe measure. However, traditional breeding methods are difficult to breed insect-resistant varieties, and the cycle is long, making it difficult to obtain breakthrough progress. The emergence of transgenic insect-resistant plants has opened up new ideas for the control of plant diseases and insect pests. The use of Bt insect-resistant tr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12P19/34C07K14/42A01P7/04
Inventor 吴志明谢晓亮陈劲枫王玉海温春秀刘铭田伟周巧梅董文琦
Owner 河北省农林科学院经济作物研究所
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