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Specific promotor in rice embryo and application thereof

A promoter and rice technology, applied in the direction of using vectors to introduce foreign genetic material, cells modified by introducing foreign genetic material, DNA/RNA fragments, etc., can solve the problem of few miRNA promoters, difficult to find promoters, and miRNA matching The problem of multiple regions

Inactive Publication Date: 2015-03-18
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some genes, such as non-structural genes, will undergo gene transfer, so it is difficult to find the corresponding promoters that are specifically expressed in specific tissues. However, due to the fact that miRNAs are too short and have too many matching regions, there are very few tissue-specific miRNA promoters found on this basis.

Method used

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  • Specific promotor in rice embryo and application thereof
  • Specific promotor in rice embryo and application thereof
  • Specific promotor in rice embryo and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1. Cloning of rice mesoembryo-specific promoter

[0068] Using the rice genomic DNA extracted by conventional methods as a template, the promoter P1 was amplified by conventional PCR using the following primer pair (SEQ ID NO.: 2 and 3):

[0069] Forward primer: GGCTGCAGCGGCTGCTGTTCTTATGGC (SEQ ID NO: 2)

[0070] Reverse primer: GGGATCCTGCACTGCAAATCCGGTTAG (SEQ ID NO: 3).

[0071] The inventors also used the same method to amplify two other promoters: P2 and P3, using the following different reverse primers:

[0072] Reverse primer: GGGATCCGCACTGCAAATCCGGTTAG (SEQ ID NO:4)

[0073] Reverse primer: GGGATCCACTGCAAATCCGGTTAG (SEQ ID NO: 5).

Embodiment 2

[0074] Example 2. Construction of GUS expression vector

[0075] The promoter sequences obtained in Example 1 were respectively connected to the commercially available vector pMD19-T vector (Takara Company), and verified by sequencing. Then use HindIII and BamHI to double digest the pMD19-T vector, reclaim the promoter fragment of about 2kb, and use the restriction endonuclease HindIII to this fragment and the binary vector pBI101.1 (Clontech Company, U.S.) containing the GUS reporter gene respectively Double enzyme digestion with BamHI, the digested product was recovered by gel, ligated with T4-DNA ligase, and transformed into Escherichia coli DH5α strain for preservation.

[0076] The final vectors obtained were named pBI101.1-P1-GUS, pBI101.1-P2-GUS and pBI101.1-P3-GUS. The recombinant vectors pBI101.1-P1-GUS, pBI101.1-P2-GUS and pBI101.1-P3-GUS were transformed into Agrobacterium strain GV3101 by heat shock transformation method, and verified by PCR amplification.

Embodiment 3

[0077] Embodiment 3. Obtaining of transgenic plants

[0078] The constructed vectors pBI101.1-P1-GUS, pBI101.1-P2-GUS and pBI101.1-P3-GUS were respectively transformed into commercially available Agrobacterium GV3101 strains, and then the wild-type rice was treated by flower dipping method. 11 for transformation.

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Abstract

The invention relates to a specific promotor in rice embryo and an application thereof. The promotor is particularly suitable for driving exogenous genes to be specifically expressed in the rice embryo and not to be expressed in other tissues of rice. The promotor is high in expression specificity. The invention also provides a structure, an expression cassette and a carrier containing the promotor. The transgenic rice containing the promotor is capable of specifically improving the agronomic trait of the rice, effectively avoiding the exogenous genes from being introduced to influence the other tissues of the rice and specifically expressing the exogenous genes in the embryo so as to obtain various related products.

Description

technical field [0001] The invention belongs to the field of biotechnology. Specifically, the present invention relates to a rice embryo-specific promoter and its application. Background technique [0002] Rice is one of the most important food crops in the world. Half of the world's population takes rice as a staple food. my country's annual rice consumption and output account for one-third of the world's total. my country is a populous country, increasing food production has always been a major social issue. The reduction of arable land area and the increase of population have intensified the requirements for rice yield and quality, and high yield and high quality have increasingly become the focus of rice research. Rice has a relatively small genome and is a model plant for functional genome research of grass crops. [0003] Embryo is formed during the sexual reproduction of angiosperms and has important biological significance in plant development. Embryo development...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/63C12N15/82C12N5/10
Inventor 时振英戴争妍
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI