Recombinant vector for rapidly obtaining non-GMO genome editing plants and method for using same

A technology of recombinant vectors and gene editing, which is applied in botany equipment and methods, genetic engineering, recombinant DNA technology, etc., can solve the problems of laborious, expensive, false negative results, time-consuming, etc., to shorten flowering time, shorten time, improve efficiency effect

Active Publication Date: 2019-01-18
YUNNAN ACAD OF TOBACCO AGRI SCI
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Problems solved by technology

For the screening of non-GMO ingredients in the T1 generation, various derivative methods based on PCR technology are generally used, but DNA extraction, specifi

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  • Recombinant vector for rapidly obtaining non-GMO genome editing plants and method for using same
  • Recombinant vector for rapidly obtaining non-GMO genome editing plants and method for using same
  • Recombinant vector for rapidly obtaining non-GMO genome editing plants and method for using same

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Embodiment Construction

[0021] The present invention will be further described in detail below in conjunction with the examples.

[0022] Quickly obtain the recombinant vector of non-transgenic gene editing plants, the recombinant vector carries NtFT and PAP1 expression elements; the recombinant vector is inserted into the original vector containing PAP1 and NtFT expression elements, the expression element is named PF Cassete, its core The nucleotide sequence is as SEQ ID No.1; the original vector is a CRISPR / Cas9 carrier for plant gene editing containing sgRNA gene, Cas9 gene and screening tag gene, and all the above-mentioned original vectors based on PF Cassete Transformations all belong to the protection scope of the present invention. The NtFT expression element of the present invention produces a protein that promotes early flowering of plants, and the PAP1 expression element produces a protein that promotes plant anthocyanin biosynthesis.

[0023] The nucleotide sequence list is as follows: ...

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Abstract

Provided are a recombinant vector for rapidly obtaining non-GMO genome editing plants and a method for using the same. The recombinant vector comprises an expression element containing PAP1 and NtFT;The original vector is a vector of CRISPR/Cas9 for plant gene editing containing sgRNA gene, Cas9 gene and screening tag gene; The NtFT expression element produces proteins that promote early flowering and the PAP1 expression element produces proteins that promote anthocyanin biosynthesis in plants. The invention can greatly shorten the time for obtaining the gene editing mutation offspring, and screens the offspring plants by plant color, thus not only retaining the offspring of the target gene mutation, but also ensuring that the mutation offspring does not contain the transgenic fragment, and simultaneously can significantly improve the gene editing efficiency. The invention can be used for preparing the gene editing mutation offspring, and can greatly shorten the time for obtaining thegene editing mutation offspring. The method of the invention is simple, fast, cheap and efficient.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant vector for obtaining non-transgenic gene editing plants and a method for using it. Background technique [0002] In 2012, clustered regularly interspaced short palindromic repeats (Clustered regularly interspaced short palindromic repeats / CRISPR-associated protein 9, CRISPR-associated protein 9, CRISPR / Cas9) and related nuclease Cas9 were reported to be used for gene editing. CRISPR / Cas9 is a new gene editing system developed after Zinc finger nuclease (ZFN) and transcription activator like effector nuclease (TALEN). After just a few years of development, it has quickly become the most successful and widely used genome editing technology because of its simple operation, high editing efficiency, and low off-target efficiency. The CRISPR / Cas9 system has been successfully applied to major food crops such as rice, wheat, corn, soybeans, and economic crops such as cotton, t...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00A01H6/82
CPCC12N15/8213C12N15/8261
Inventor 黄昌军刘勇于海芹曾建敏李永平
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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