Constitutive promoters and their uses

A constitutive promoter, plant tissue technology, used in applications, recombinant DNA technology, biochemical equipment and methods, etc.

Active Publication Date: 2015-11-18
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the ubiquitin gene promoters from plants are mainly isolated from maize, sunflower, tobacco, potato, sugarcane, Arabidopsis and soybean, but the ubiquitin gene promoters in rice are rarely reported

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

[0060] The first embodiment, the synthesis of prOsRUBQ1 promoter sequence

[0061] Obtain the constitutive promoter prOsRUBQ1 of the present invention, as shown in SEQIDNO: 1 in the sequence listing; the prOsRUBQ1 promoter sequence (as shown in SEQIDNO: 1 in the sequence listing) was synthesized by Nanjing GenScript Biotechnology Co., Ltd.; the synthesized The 5' end of the prOsRUBQ1 promoter sequence (SEQ ID NO: 1) is also connected with a KpnII restriction site, and the 3' end of the prOsRUBQ1 promoter sequence (SEQ ID NO: 1) is also connected with a HindIII restriction site.

no. 2 example

[0062] The second embodiment, construction of recombinant expression vector p100024 containing prOsRUBQ1 promoter sequence and transformation of recombinant expression vector into Agrobacterium

[0063] 1. Construction of the recombinant cloning vector pT-prOsRUBQ1 containing the prOsRUBQ1 promoter sequence

[0064] The prOsRUBQ1 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-prOsRUBQ1. The construction process is as follows: figure 1 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of phage f1; LacZ is the LacZ start codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; prOsRUBQ1 is the rice variety Nipponbare Ubiqutin (ubiquitin) 1 gene promoter (SEQ ID NO: 1); MCS is a multiple clon...

no. 3 example

[0075] The third embodiment, the acquisition of maize plants transferred to the prOsRUBQ1 promoter sequence

[0076] According to the routinely used Agrobacterium infection method, the immature embryos of the aseptically cultivated corn variety Zong 31 (Z31) were co-cultured with the Agrobacterium described in 4 in the second example, so as to infect 2 and 3 in the second example. The T-DNA (including the promoter sequence of maize Ubiquitin gene, prOsRUBQ1 promoter sequence, control sequence, PAT gene, PMI gene and Nos terminator sequence) in the constructed recombinant expression vector p100024 and p100079 (control sequence) was transferred into maize In the genome, maize plants transformed with the prOsRUBQ1 promoter sequence and maize plants transformed with the control sequence (positive control) were obtained; meanwhile, wild-type maize plants were used as negative controls.

[0077] For Agrobacterium-mediated maize transformation, briefly, immature immature embryos are ...

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Abstract

The invention relates to a constitutive promoter and application thereof. The nucleotide sequence of the constitutive promoter comprises the sequence of (a) or (b): (a) the nucleotide sequence as shown in SEQ (sequence) ID (identity) NO: 1; (b) the nucleotide sequence which is hybridized with the nucleotide sequence limited by (a) under strict conditions. The constitutive promoter provided by the invention almost generally has activity in most of tissues and many types of cells of plants, and can significantly improve the expression level of a target heterologous nucleotide sequence in the tissues of the plants, in particular in leaf blades and stems.

Description

technical field [0001] The invention relates to a constitutive promoter and its use, in particular to a constitutive rice ubiquitin gene promoter and its use. Background technique [0002] Major advances in cell biology and transgenic technology have allowed the incorporation of foreign genes into many crops. Foreign genes are transferred into plants mainly to express proteins, which are called "transgenic plants", and the proteins will impart beneficial traits, such as resistance to pathogenic microorganisms or insects, resistance to herbicides, tolerance to drought or other unfavorable surroundings. Usually, the DNA coding region of the foreign gene is linked to a strong and constitutive DNA promoter region to ensure efficient expression of the foreign gene in transgenic cells. [0003] A large number of common promoters are used to drive the expression of foreign genes in transgenic plants. The cauliflower mosaic virus (CaMV) 35S promoter is widely used in both dicots ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/84
Inventor 张成伟丁德荣王登元吴业春庞洁
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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