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Method for cloning promoter of specific expression gene EXP18D of paddy rice root

A technology for expressing genes and cloning methods, applied in the field of cloning of rice root-specific expression gene EXP18D promoter

Inactive Publication Date: 2013-06-12
CHINA AGRI UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

Most of the plant functional genes such as hormone synthesis, metabolism-related genes, plant growth and development regulation genes are expressed in the above-ground and underground parts of plants, and the simple transgenic or gene silencing methods can only regulate the genes in plants as a whole. expression, cannot specifically regulate the function and development of plant roots

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  • Method for cloning promoter of specific expression gene EXP18D of paddy rice root
  • Method for cloning promoter of specific expression gene EXP18D of paddy rice root
  • Method for cloning promoter of specific expression gene EXP18D of paddy rice root

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Embodiment 1

[0038] A method for cloning a rice root-specific expression gene EXP18D promoter, the method comprising the following steps:

[0039] Ⅰ. Obtaining Arabidopsis root-specific genes:

[0040] A. Extract total RNA from Arabidopsis thaliana root and total RNA from Arabidopsis thaliana leaves from Arabidopsis thaliana plants, and then carry out reverse transcription PCR reaction respectively, and amplify the cNDA sequence of Arabidopsis thaliana root and Arabidopsis thaliana leaf respectively ;

[0041] The specific operation is:

[0042] The roots and leaves of Arabidopsis thaliana plants grown for 20 days were taken as materials, and the total RNA of Arabidopsis roots and leaves were extracted by Trizol method, and reverse transcription PCR reactions were carried out to obtain Arabidopsis root cDNA and Arabidopsis thaliana root cDNA respectively. leaf cDNA;

[0043] The system of the reverse transcription PCR reaction is: RNA: 1.2 μg, Oligo(dT)15: 2 μl,

[0044] M-MLV buffer: ...

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Abstract

The invention which belongs to the plant genetic engineering field provides a method for cloning a promoter of a specific expression gene EXP18D of a paddy rice root. The method is characterized in that: a specific gene EXP18 of Arabidopis thaliana roots is firstly obtained, and then the promoter of the specific expression gene EXP18D of the paddy rice root is obtained. The method allows the promoter of the specific expression gene EXP18D of the paddy rice root to be cloned in the invention, and the promoter which is a promoter for the specific expression of plant roots can drive genes with relevant functions to express, so the promoter has potentials for specifically changing the form of the plant roots and obtaining corresponding drought resistant plants.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to a method for cloning a rice root-specific expression gene EXP18D promoter. Background technique [0002] The promoter is the most important factor in the regulation of gene expression, which basically determines whether, when and where a gene is expressed. According to the mode of action and function, promoters can be roughly divided into three categories: constitutive promoters, specific promoters and inducible promoters. At present, the regulatory mechanism of tissue-specific promoters has entered the stage of in-depth research. In addition to the basic structure, such promoters usually have some sequences that regulate specific expression. These sequences provide binding sites for transcription factors. interaction to regulate gene expression. Driven by tissue-specific promoters, gene expression usually only occurs in specific tissues or organs, and often exhibits de...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 贾文锁梁莉艳赵艳侠李自超李金杰李冰冰
Owner CHINA AGRI UNIV