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Method for cloning arabidopsis root specificity expressed gene ARSK1 promoter

An Arabidopsis thaliana root and gene expression technology, applied in the field of plant genetic engineering

Inactive Publication Date: 2012-04-18
BEIJING AGRO BIOTECH RES CENT
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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 arabidopsis root specificity expressed gene ARSK1 promoter
  • Method for cloning arabidopsis root specificity expressed gene ARSK1 promoter
  • Method for cloning arabidopsis root specificity expressed gene ARSK1 promoter

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Embodiment 1

[0033] A method for cloning the Arabidopsis root-specific expression gene ARSK1 promoter, the method comprising the following steps:

[0034] Ⅰ. Obtain the Arabidopsis root-specific gene ARSK1:

[0035] 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 ;

[0036] The specific operation is:

[0037] 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;

[0038] The system of the reverse transcription PCR reaction is: RNA: 1.2 μg, Oligo(dT)15: 2 μl,

[0039] ...

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Abstract

The invention relates to the field of plant genetic engineering, in particular to a method for cloning an arabidopsis root specificity expressed gene ARSK1 promoter. The method comprises the following steps of: obtaining an arabidopsis root specificity expressed gene ARSK1 and obtaining the arabidopsis root specificity expressed gene ARSK1 promoter. The method provided by the invention can be used for cloning the arabidopsis root specificity expressed gene ARSK1 promoter, the promoter, which is expressed by plant root specificity, can be used for driving the expression of related function gene, so that the promoter has the capacity of specifically changing root form of the plant and obtaining corresponding drought-resisting plant; and the livability of the promoter with the plant being transferred in is 60 percent to 70 percent higher than that of a wild type plant of a control group at same drought conditions.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to a method for cloning the ARSK1 promoter of the root-specific expression gene of Arabidopsis thaliana. 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 or...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/113
Inventor 吴忠义黄丛林张秀海罗昌程曦
Owner BEIJING AGRO BIOTECH RES CENT