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MYB transcription factor PtrMYB01 in Populus tomentosa Carr and cloning method of cDNA of PtrMYB01and application thereof

A technique of transcription factor and cloning method, which is applied in the field of MYB transcription factor and its cDNA cloning, and can solve the problem of low expression

Inactive Publication Date: 2012-04-04
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, BcMYB1 in Heliconia pachyrhiza is strongly induced by drought, and can respond to PEG, high-salt, low temperature and other stresses to a certain extent, but BcMYB1 in exogenous abscisic acid (ABA), methyl jasmonate (MeJa), Under the treatment of salicylic acid (SA), its expression level was very low, indicating that it may participate in the regulation of gene expression through an ABA-independent pathway to respond to drought

Method used

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  • MYB transcription factor PtrMYB01 in Populus tomentosa Carr and cloning method of cDNA of PtrMYB01and application thereof
  • MYB transcription factor PtrMYB01 in Populus tomentosa Carr and cloning method of cDNA of PtrMYB01and application thereof
  • MYB transcription factor PtrMYB01 in Populus tomentosa Carr and cloning method of cDNA of PtrMYB01and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0117] Example 1: Populus tomentosa Carr, existing materials in the laboratory.

[0118] 1. Total RNA extraction:

[0119] ①. Take 5g of plant tissue and add liquid nitrogen to fully grind it, transfer it to a pre-cooled 50mL centrifuge tube, add in order: 15mL guanidine isothiocyanate solution, 1.5mL 2M NaAc (pH4.0), 15mL water-saturated phenol and 3 mL chloroform / isoamyl alcohol. After mixing, place on ice for 15 min.

[0120] ②, 4°C, centrifuge at 15000g for 30min, transfer the supernatant to another tube, add an equal volume of isopropanol, mix well and precipitate at -20°C for 1hr.

[0121] ③. Centrifuge at 15,000 g for 25 min at 4°C, discard the supernatant, add 5 mL of guanidine isothiocyanate solution to dissolve the precipitate, and then add isopropanol at -20°C for 1 hr. The RNA precipitate was collected by centrifugation, washed once with 70% ethanol and dried, and 500 μl of DEPC-treated water-soluble RNA was added. Take 5 μl of electrophoresis to check the inte...

Embodiment 2

[0169] The Agrobacterium DHA105 obtained in Example 1 is used to infect plants to obtain transgenic plants, which may increase their stress resistance (such as cold resistance and drought resistance), and can be used in fields or forests.

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Abstract

The invention provides a novel MYB transcription factor PtrMYB01 in Populus tomentosa Carr, a gene coding the transcription factor, and a cloning method of cDNA, wherein, the transcription factor is polypeptide which is obtained by the separation of Populus tomentosa Carr. By analyzing a highly homologous transcription factor ATMYB60 which belongs the same family with the transcription factor PtrMYB01, the transcription factor PtrMYB01 has possible functions of regulating and controlling plant stress resistance (drought resistance) and regulating anabolism of plant flavonoids. Accordingly, if the recombinant plasmid of the PtrMYB01 gene is transferred into the plant, because the gene is overexpressed in the plant, a drought-resistant cold-resistance plant can be produced, so that the plant yield is increased. The cloning method comprises the following steps: extracting total RNA from Populus tomentosa Carr, and carrying out inverse transcription on the total RNA to synthesize cDNA; designing primers according to the conserved sequence of the transcription factor, and amplifying into a DNA fragment through chain polymerization enzyme reaction; purifying the amplified products, cloning the purified products to a pCXSN vector, transferring a DH5 alfa cell, and culturing overnight in plate. The transferred escherichia coli plasmid is extracted and transferred into Agrobacterium EHA105 for subsequent research.

Description

technical field [0001] The invention belongs to the technical field of biological gene engineering, relates to the cloning technology in the technical field of biological gene, and specifically relates to the cloning of a new MYB type transcription factor and its cDNA in Populus tomentosa. Background technique [0002] Populus tomentosa Carr is a plant of the genus Populus in the family Salicaceae, a deciduous tree with a height of 30 meters and a diameter at breast height of 2 meters. It is a unique native tree species in my country. Populus tomentosa is distributed in a wide range, from southern Liaoning and Inner Mongolia in the north to the Yangtze River Basin in the south, with the middle and lower reaches of the Yellow River as the suitable habitat. Poplar tomentosa has a tall trunk, strong adaptability, and long fibers. It is an important civil and industrial wood. In addition, Populus tomentosa has the medicinal effect of clearing heat and dampness, relieving cough ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/10A01H5/00
Inventor 王晴岚秦智超刘松领罗克明
Owner SOUTHWEST UNIVERSITY
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