Efficient marking method for heavy ion radiation mouseearcress genome target spots

A technology of heavy ion irradiation and Arabidopsis thaliana, which is applied in the field of efficient labeling of Arabidopsis genome targets by heavy ion irradiation, can solve the problems of cumbersome experimental process, low labeling efficiency, and complexity, and achieve simple experimental operation and high labeling efficiency Effect

Inactive Publication Date: 2019-05-21
HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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Problems solved by technology

However, the shortcomings of the above two experimental systems are: 1) the experimental process is cumbersome and complicated; 2) the labeling efficiency is low; 3) non-DSB-specific induction

Method used

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  • Efficient marking method for heavy ion radiation mouseearcress genome target spots
  • Efficient marking method for heavy ion radiation mouseearcress genome target spots
  • Efficient marking method for heavy ion radiation mouseearcress genome target spots

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Experimental program
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Effect test

Embodiment 1

[0037] High-efficiency labeling method for genomic targets in Arabidopsis thaliana irradiated by heavy ions

[0038] (1) Experimental materials: seeds of the Arabidopsis homologous recombination reporter gene line irradiated with 30Gy of heavy ions;

[0039] The Arabidopsis homologous recombination reporter line R3L66 introduces completely homologous and inverted repeat intron fragments into the reporter gene GUS to form a substrate sequence for homologous recombination to inactivate the reporter gene, and the recombination substrate sequence of the reporter gene After genetic damage, restore the activity of the reporter gene through homologous recombination repair, the specific principle is as follows figure 1 shown;

[0040] The size of the intron fragment is 589bp, and the sequence of the intron is shown in SEQ NO.1.

[0041] (2) Experimental steps:

[0042] (1) The irradiated seeds were sown in MS medium after 2 days of vernalization, the MS medium contained 1% sucrose,...

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Abstract

The invention discloses an efficient marking method for heavy ion radiation mouseearcress genome target spots, and relates to the field of heavy ion plant radiation induction breeding mechanism research. In order to solve the problem that an existing reporter gene has defects for marking heavy ion radiation plant genome target spots, the method comprises the steps that a mouseearcress homologous recombination report system is adopted as an experimental material, the heavy ion radiation plant genome target spots are efficiently marked in the mode that the activity of the reporter gene is recovered after the reporter gene is damaged by DNA dual-chain breakage and subjected to homologous recombination repair, red recombination fragment and a flanking genome sequence thereof are amplified through PCR high fidelity, the sequencing detection amplification sequence basic group mutation condition is cloned, and therefore the heavy ion radiation plant genome target spots are marked. The methodhas the advantages that experiment operations are easy, the marking efficiency is high, and specific induction of DNA dual-chain breakage is achieved.

Description

technical field [0001] The invention relates to the field of heavy ion plant radiation mutation breeding mechanism research, in particular to a high-efficiency labeling method for heavy ion radiation Arabidopsis genome targets. Background technique [0002] Mutation breeding is a breeding method that uses physical and chemical mutagens to induce plant genetic variation and obtain excellent genetic variation resources in a short period of time. Mutation breeding can not only significantly increase the frequency of gene mutation, but also effectively broaden the type of variation (generate new genes), enriching the "gene pool" of crops. Radiation mutation breeding started in the 1920s. In 1927, Muller used X-rays to induce a large number of mutations in fruit flies. The next year, Stadler proved that X-rays can induce mutations in corn and barley. In the following 90 years, more than 60 countries have bred more than 3,200 new crop varieties using radiation mutation breeding t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895
Inventor 卞坡王婷吴京京
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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