Promoter capable of driving expression of gene in tissue of flower

A promoter and expression vector technology, applied in the field of promoters, can solve the problems of normal elongation of filaments, hindered stamen development, male sterility, etc., achieve broad application space and market prospects, and promote development and growth.

Active Publication Date: 2012-07-25
TSINGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It was previously reported that Arabidopsis thaliana MYB21 and MYB24 are related to the development of flowers. After the deletion of MYB21 and MYB324 genes in Arabidopsis, the development of stamens is blocked, and filaments cannot be elongated normally, resulting in male sterility.

Method used

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  • Promoter capable of driving expression of gene in tissue of flower
  • Promoter capable of driving expression of gene in tissue of flower
  • Promoter capable of driving expression of gene in tissue of flower

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Example 1, the acquisition of MYB21pro sequence

[0026] Using Arabidopsis thaliana (Columbia-0) leaf DNA as a template, using a primer pair composed of F1 and R1, PCR amplification was performed under the action of high-fidelity PFU enzyme to obtain PCR amplification products.

[0027] F1:5'-agaAAGCTTataattactccacctt-3':

[0028] R1: 5'-tttTCTAGAtttagagagaggaatat-3'.

[0029] The PCR amplification product was sequenced, as shown in sequence 1 of the sequence listing. The DNA shown in Sequence 1 of the Sequence Listing was named MYB21 promoter (MYB21pro).

Embodiment 2

[0030] Embodiment 2, the acquisition and identification of transgenic plants

[0031] 1. Construction of recombinant expression vector (pBI121-MYB21pro-GUS)

[0032] 1. The PCR amplification product obtained in Example 1 was double-digested with restriction endonucleases HindIII and XbaI to obtain the digested product.

[0033] 2. The pBI121 vector (GenBank: AF485783.1) was double digested with restriction endonucleases HindIII and XbaI, and the vector backbone of about 14 kb was recovered.

[0034] 3. Ligate the digested product of step 1 and the vector backbone of step 2 under the action of T4 DNA ligase to obtain the recombinant plasmid pBI121-MYB21pro-GUS. The schematic diagram of the structure of the recombinant plasmid pBI121-MYB21pro-GUS is shown in figure 1 . According to the sequencing results, the structure of the recombinant plasmid pBI121-MYB21pro-GUS is described as follows: the recombinant plasmid obtained by inserting the DNA shown in sequence 1 in the sequen...

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Abstract

The invention discloses a promoter capable of driving the expression of a gene in a tissue of flower. The promoter provided by the invention is a DNA (Deoxyribonucleic Acid) molecule as described in 1), 2) or 3): 1) the DNA molecule as shown in a sequence 1 of a sequence list; 2) the DNA molecule being hybridized with a DNA sequence defined in 1) under strict conditions and having the functions of the promoter; and 3) the DNA molecule having over 90% homology with the DNA sequence defined in 1) or 2) and having the functions of the promoter. The promoter provided by the invention can be used for driving the expression of an exogenous gene in the tissue of the flower. The promoter is helpful for promoting study of people on the development and growth of Arabidopsis, provides an economic, quick and effective approach for the genetic alteration of plants (in particular cruciferous vegetable and flower), and has broad application space and market prospect in the agricultural field.

Description

technical field [0001] The present invention relates to a promoter capable of driving gene expression in flower tissue. 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] MYB factors are a family of proteins that contain a conserved MYB DNA-binding domain. Typical MYB factors are related to c-Myb, and play a regulatory role in the cell cycle of animals, plants and some higher eukaryotes. There are at least 100 R2R3-MYB genes in Arabidopsis. These genes play an important role in regulating plant secondary metabolism, maintaining cell shape, resisting diseases, and responding to hormones. It has been previously reported that Arabidopsis MYB21 and MYB24 are related to flower development. After the deletion of MYB21 and MYB324 genes in Arabidopsis, the development of stamens is blocked, and filaments cannot b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113
Inventor 谢道昕黄煌宋素胜齐天从吴德伟彭文樊萌
Owner TSINGHUA UNIV
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