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Plant DREB transcription factor and its coding gene and use

A technology of transcription factors and coding genes, applied in the field of plant biology, can solve problems such as the comprehensive improvement of stress resistance of difficult plants, and achieve the effects of extensive practical significance and far-reaching theoretical significance

Active Publication Date: 2007-05-02
北京元熙投资有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the stress tolerance of plants is a complex trait regulated by multiple genes, it is difficult to ac

Method used

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  • Plant DREB transcription factor and its coding gene and use
  • Plant DREB transcription factor and its coding gene and use
  • Plant DREB transcription factor and its coding gene and use

Examples

Experimental program
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Embodiment 1

[0031] Embodiment 1, the acquisition of DMY-DREB and its coding gene

[0032] Rice was extracted by CTAB method (Steiner JJ, Poklemba CJ, Fjellstrom RG, Elliott LF., A rapidone-tube genomic DNA extraction process for PCR and RAPD analyses. Nucleic Acids Res. 1995 Jul 11; 23(13): 2569-70.) Grass genome DNA, the specific method is as follows: take Spartina leaves, grind with liquid nitrogen, add 5 ml of CTAB solution preheated at 68°C (add 10mM mercaptoethanol before use), and heat at 60°C for 30 minutes per 1 gram of plant material . Then add an equal volume of chloroform:isoamyl alcohol, centrifuge at 15000g for 15 minutes, and take the supernatant. Add 2 / 3 times the volume of isopropanol to the supernatant, mix well, use the needle tip of the injection needle to stir out the DNA filament and place it in a new tube, then add 75% ethanol to wash the precipitate and the wall of the centrifuge tube, and then add ethanol Blot dry, air dry remaining ethanol, add TE to dissolve DN...

Embodiment 2

[0036] Embodiment 2, the construction of DMY-DREB plant expression vector and the stress tolerance experiment of transgenic tobacco

[0037] The Arabidopsis rd29A gene promoter sequence was amplified by Rd29A-S and Rd29A-A PCR methods using CTAB method to extract Arabidopsis genomic DNA as a template. Among them, Rd29A-S: 5'-TCTAGACGCATGATTTGATGGAGGAG-3', Rd29A-A: 5'-TCCAAAGATTTTTTTCTTTCCAA-3'. 25ul PCR reaction system contains 100ng genomic DNA, 1.5mM MgCl 2 , 20 mM Tris-HCl (pH8.4), 50 mM KCl, 0.8 mM dNTP mixture, 1 μM Rd29A-S and 1 μM Rd29A-A, 1 U of Pfu enzyme (Shanghai Shenyou Biotechnology Company). Carry out PCR cycling in a PCR-thermal cycler (Eppendorf, Germany) according to the following scheme: first 94°C for 5 minutes; then 94°C for 1 minute, 54°C for 1 minute, 72°C for 1 minute, a total of 30 cycles; finally 72°C 10 minutes. Use the gel recovery kit (Beijing Tianwei Times Technology Co., Ltd.) to recycle the amplified DNA fragment of about 944bp according to th...

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Abstract

The invention discloses a plant DREB transcriptional factor, the coding gene and the application. It is the albumen that contains one of the following amino acid residue gene sequences: 1) SEQ ID: No:1 in the sequence table; 2)the albumen relative to the plant anti-reverse ability and is taken placed, absented or added by one or several amino acid residues in SEQ ID No:1. The invention could enhance the endurance of the plants.

Description

technical field [0001] The invention relates to a plant DREB transcription factor and its coding gene and application in the field of plant biotechnology, in particular to a DREB transcription factor related to stress tolerance derived from Spartina anglica, its coding gene and its application. Background technique [0002] Under adversity stress, plants will produce a series of responses, accompanied by many physiological, biochemical and developmental changes. Elucidating the response mechanism of plants to stress will provide a scientific theoretical basis for molecular breeding of plant stress resistance. [0003] Many genes in plants have been shown to be induced by abiotic stresses. Stress-related gene products can be divided into two categories: the first category of gene-encoded products include ion channel proteins, aquaporins, osmotic regulators (sucrose, proline and betaine, etc.) Responsive gene products; the second type of gene-encoded products include protein...

Claims

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Application Information

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IPC IPC(8): C07K14/415C07H21/04C12N15/29C12N15/63C12N15/82
Inventor 陈蕾徐建勇
Owner 北京元熙投资有限公司