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Aquaporin gene promoter for chorispora bungeana and cloning method thereof

A technology of alpine ion mustard and channel protein, applied in the field of alpine ion mustard aquaporin gene promoter and its cloning, which can solve the problems of no reports

Inactive Publication Date: 2015-02-04
李鸣
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the research on the promoter of the alpine ion mustard aquaporin gene

Method used

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  • Aquaporin gene promoter for chorispora bungeana and cloning method thereof
  • Aquaporin gene promoter for chorispora bungeana and cloning method thereof
  • Aquaporin gene promoter for chorispora bungeana and cloning method thereof

Examples

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Embodiment 1

[0021] Take 10 μg of total genomic DNA from mustard root, digest with restriction endonuclease Tail (MaeII*) at 65°C overnight, and stop the digestion reaction at 80°C for 20min. Dilute DNA concentration to 2ng / L, use T 4 DNA ligase performs self-cyclization reaction, using the cyclization product as a template, and performing nested PCR amplification with specific primers N2 / N3 and N1 / N4, respectively, to obtain a single sequence of about 800bp, T-cloning, and sequencing.

[0022] Referring to the operation manual of the Genome Walking Kit (TaKaRa) kit, 3 primers SP1, SP2, and SP3 were designed according to the full-length gene sequence of ChlRB7. Three rounds of nested PCR amplification were carried out sequentially with the total genomic DNA of Alpine mustard as a template, and a single fragment of 1282bp was obtained, which was sequenced by T-cloning and sequenced. The obtained fragment was highly homologous to the previous 800bp fragment sequence, and further analysis de...

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Abstract

The invention relates to an aquaporin gene promoter for chorispora bungeana and a cloning method thereof. The promoter is an important cis element for regulation of gene expression and is further an important element for a gene engineering expression vector. Therefore, the cloning method of the aquaporin gene promoter for chorispora bungeana provided by the invention plays an important role in expression regulation of chorispora bungeana.

Description

technical field [0001] The invention relates to an alpine mustard aquaporin gene promoter and a cloning method thereof, belonging to the field of plant stress resistance gene research. Background technique [0002] A promoter is a DNA sequence used for the positioning of RNA polymerase, usually located upstream of a gene. Once RNA polymerase locates and binds to the promoter, the transcription process can be initiated, so the promoter is an important cis-element for the regulation of gene expression, and its interaction with trans-elements such as RNA polymerase and other protein cofactors is the promoter. The essence of the regulatory model. [0003] The final products of gene expression are RNA and protein, these two types of substances and their secondary products maintain the orderly activities of the whole life. Any defect or disorder in the regulation of gene expression will cause serious consequences to the organism. Therefore, the mechanism of gene expression regu...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/10
Inventor 李鸣高天鹏张勇晋玲刘光琇安黎哲
Owner 李鸣
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