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Expression carrier of double-valence anti-disease gene in plant

A plant expression vector and disease-resistant gene technology, applied in the field of plant genetic engineering, to achieve the effect of improving sustained disease resistance

Inactive Publication Date: 2005-03-23
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the genes commonly used in plant disease resistance genetic engineering often have strong disease resistance specificity, and most of them are single gene transformations, so they can only control a certain pathogen or physiological race and strain, and pathogenic bacteria are more likely to develop resistance

Method used

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  • Expression carrier of double-valence anti-disease gene in plant

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0013] Construction of plant expression vector pBin35SGAFP-NPI for bivalent disease resistance gene

[0014] 1. The 35S-NP1-nos fragment obtained by digesting pBin35SNP1 with EcoRI and HindIII was connected to the pDM-302-FU vector fragment digested with EcoRI and HindIII to obtain pDM302-FU-NP1. A band of about 1.3kb was identified by EcoRI and HindIII digestion, which was 35S-NP1-nos.

[0015] 2. The 35S-NP1-nos fragment obtained by digesting pDM302-FU-NP1 with EcoRI and XhoI was connected to the pGEM-7Zf vector fragment digested with EcoRI and XhoI to obtain pGEM-NP1b. The positive clone obtained by blue-white screening was passed through EcoRI , XhoI double enzyme digestion identification, a band of about 1.3kb can be seen, namely 35S-NP1-nos.

[0016] 3. The 35S-NP1-nos fragment obtained by digesting pGEM-NP1b with HindIII was connected to the pBin35SGAFP vector fragment digested with HindIII to obtain pBin35SGAFP-NP1, and the 1.3kb band was cut by HindIII, indicating 35...

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Abstract

The invention discloses a divalent disease-resistant gene plant expression carrier, containing starter, rabbit alexin gene, Gastrodiae anti-fungal protein gene, terminator and screening label gene, all expressed in composition form in plant cell. And its characteristic: it recombines rabbit alexin gene and Gastrodiae anti-fungal protein gene with plant cell expression carrier. Its transgenic plant cell will contain rabbit alexin protein and Gastrodiae anti-fungal protein. It contains two wider antibacterial spectrums of disease-resistant gene, but their antibacterial mechanisms are different.

Description

1. Technical field [0001] The invention relates to a plant genetic engineering, in particular to a plant expression carrier of a bivalent disease resistance gene (GAFP+NP-1). 2. Background technology [0002] Rabbit defensins belong to α-defensins, and the α-defensins have inhibitory effects on Gram-negative bacteria, positive bacteria, mycobacteria, fungi, enveloped viruses, and HIV viruses to varying degrees. Among them, NP-1 has a wide antibacterial spectrum and strong antibacterial activity, and because of its unique resistance mechanism, microorganisms are not easy to develop resistance to it. Therefore, rabbit defensin genes have extremely wide application prospects and high potential in plant disease resistance breeding. Value. Gastrodia elata antifungal protein is isolated and purified from Gastrodia elata tubers, and the antibacterial results in vitro show that it has certain resistance to more than 20 kinds of fungi. The results of prokaryotic expression and tran...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/29C12N15/65C12N15/82
Inventor 陈英王义琴黄敏仁孙勇如王光萍张新叶王明庥诸葛强徐立安
Owner NANJING FORESTRY UNIV
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