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Plant promoter and application thereof

A promoter and plant technology, applied in the field of plant promoters, can solve problems such as affecting gene expression level and affecting affinity, and achieve the effect of promoting development and growth, broad application space and market prospects

Active Publication Date: 2013-09-25
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The initiation of transcription is a key stage of gene expression, and an important issue at this stage is the interaction between RNA polymerase and promoter: the structure of the promoter affects its affinity with RNA polymerase, thereby affecting the level of gene expression

Method used

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  • Plant promoter and application thereof
  • Plant promoter and application thereof
  • Plant promoter and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1. Obtaining of bHLH14pro sequence

[0021] Using the genomic DNA of leaves of Arabidopsis thaliana ecotype Columbia as a template, a DNA molecule with a promoter function was found through PCR amplification and sequencing, as shown in sequence 1 of the sequence listing. The DNA molecule shown in sequence 1 of the sequence listing is named bHLH14 promoter (also known as bHLH14pro).

Embodiment 2

[0022] Embodiment 2, the acquisition and identification of transgenic plants

[0023] 1. Construction of recombinant expression vector

[0024] 1. Using the genomic DNA of the leaves of the Columbia ecotype Arabidopsis thaliana as a template, a primer pair composed of F1 and R1 is used for PCR amplification, and the PCR amplification product is recovered.

[0025] F1: 5'-aa ctgcag cctatgtcgatatgcgccgagaaggac-3';

[0026] R1: 5'-cgc ggatcc ctttccagtgatcccgaccaaagaga-3'.

[0027] 2. Double-digest the PCR amplification product of step 1 with restriction endonucleases PstI and BamHI, and recover the digested product.

[0028] 3. The pCAMBIA1391z vector was double-digested with restriction enzymes PstI and BamHI, and the vector backbone of about 14 kb was recovered.

[0029] 4. Ligate the digested product of step 2 with the vector backbone of step 3 to obtain the recombinant plasmid pCAMBIA1391z-bHLH14pro-GUS. According to the sequencing results, the structure of the recomb...

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Abstract

The invention discloses a plant promoter and application thereof. The invention provides a DNA (deoxyribonucleic acid) molecule which is as shown in (1) or (2) or (3), (1): a DNA molecule shown in a sequence 1 in a sequence table; (2): a DNA molecule which is hybridized with the DNA sequence limited in (1) under the strict condition and has a promoter function; (3): a DNA molecule which has homology of above 90 percent with the DNA sequence limited in (1) or (2) and has a promoter function. The invention provides a promoter, which is capable of driving a gene to be expressed in a plant. The invention provides an economic, rapid and effective approach for genetic modification of plants, especially vegetables and flowers of cruciferae. The plant promoter is wide in application space and market prospect in the agricultural field.

Description

technical field [0001] The present invention relates to a plant promoter and its application. Background technique [0002] Arabidopsis thaliana (Arabidopsis thaliana) is a cruciferous plant widely used in the research of plant genetics, developmental biology and molecular biology. A typical model plant. [0003] The promoter is a DNA sequence located in the upstream region of the 5' end of the structural gene, which can activate RNA polymerase to accurately combine with the template DNA and have the specificity of transcription initiation. The initiation of transcription is a key stage of gene expression, and an important issue at this stage is the interaction between RNA polymerase and promoter: the structure of the promoter affects its affinity with RNA polymerase, thereby affecting the level of gene expression . Contents of the invention [0004] The purpose of the present invention is to provide a plant promoter and its application. [0005] The present invention ...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/63C12N5/10C12N1/21C12N15/82C12N15/84A01H5/00C12N15/11
Inventor 谢道昕樊萌齐天从宋素胜
Owner TSINGHUA UNIV
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