Constitutive promoters and their uses

A promoter and constitutive technology, applied in the direction of application, recombinant DNA technology, using vectors to introduce foreign genetic material, etc., can solve the problem of low efficiency

Active Publication Date: 2015-12-30
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cauliflower mosaic virus (CaMV) 35S promoter is widely used in both dicots and monocots, but its efficiency in monocots is significantly lower than that in dicots

Method used

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  • Constitutive promoters and their uses
  • Constitutive promoters and their uses
  • Constitutive promoters and their uses

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0059] The first embodiment, the synthesis of prBrTsf1 promoter sequence

[0060] Obtain the constitutive promoter prBrTsf1 of the present invention, as shown in SEQ ID NO: 1 in the sequence listing; the prBrTsf1 promoter sequence (shown in SEQ ID NO: 1 in the sequence listing) was synthesized by Nanjing GenScript Biotechnology Co., Ltd.

no. 2 example

[0061] The second embodiment, the construction of the recombinant expression vector DBN100040 containing the prBrTsf1 promoter sequence and the transformation of the recombinant expression vector into Agrobacterium

[0062] 1. Construction of the recombinant cloning vector pT-prBrTsf1 containing the prBrTsf1 promoter sequence

[0063] The prBrTsf1 promoter sequence was connected to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector produced by Promega Company to obtain the recombinant cloning vector pT-prBrTsf1.

[0064] Then, the recombinant cloning vector pT-prBrTsf1 was transformed into Escherichia coli T1 competent cells (Transgen, Beijing, China, CAT: CD501) by heat shock method. The heat shock conditions were: 50 μl E. coli T1 competent cells, 10 μl plasmid DNA (recombinant Cloning vector pT-prBrTsf1), water bath at 42°C for 30 seconds; shake culture at 37°C for 1 hour...

no. 3 example

[0074] The third embodiment, the acquisition of soybean plants transferred to the prBrTsf1 promoter sequence

[0075] According to the commonly used Agrobacterium infection method, the cotyledon node tissue of the aseptically cultured soybean variety Zhonghuang 13 was co-cultured with the Agrobacterium described in 4 in the second embodiment to construct 2 and 3 in the second embodiment The T-DNA in the recombinant expression vector DBN100040 and DBN100036 (including eFMV: prBrTsf1 nucleotide sequence, eFMV: prAtTsf1 nucleotide sequence, CTP2 nucleotide sequence, EPSPS gene, E9 terminator sequence, prCaMV35S promoter sequence, PAT Gene and tCaMV35S terminator sequence) were transferred into the soybean genome, and soybean plants transferred to the prBrTsf1 promoter sequence and soybean plants transferred to the prAtTsf1 promoter sequence were obtained; at the same time, wild-type soybean plants were used as controls.

[0076] For Agrobacterium-mediated soybean transformation, ...

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Abstract

The invention relates to a constitutive promoter and application thereof. The nucleotide sequence of the constitutive promoter comprises the following sequence (a) or (b): (a) nucleotide sequence disclosed as SEQ ID NO:1; or (b) nucleotide sequence crossbred with the nucleotide sequence defined in (a) under strict conditions. The constitutive promoter generally has activity in most tissues and many types of cells of plants, and obviously enhances the expression level of the target heterologous nucleotide sequence in plant tissues especially in leaves and stems.

Description

technical field [0001] The invention relates to a constitutive promoter and its application, in particular to a constitutive rapeseed Tsf1 gene promoter and its application. Background technique [0002] Recent advances in plant genetic engineering have opened new doors to the engineering of improved plant traits such as plant disease resistance, insect resistance, herbicide tolerance, increased yield, improved nutritional quality of edible parts of plants and enhanced plant growth. The shelf life or stability of the final consumer product obtained. Therefore, target genes with molecular functions can be properly integrated into plant genomes to impart different or improved traits or quality. The coding sequence of the newly integrated gene is expressed in plant cells to display the novel trait of interest. It is important that appropriate regulatory signals be present in the proper structure to obtain expression of the coding sequence of the newly inserted gene in plant c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/29C12N15/82
CPCY02A40/146
Inventor 张成伟庞洁王登元刘海利康越景陶青
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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