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Analysis method for activity of mismatch repair of zebra fish embryo DNA and application thereof

A technology of zebrafish embryos and analysis methods, applied in the analysis of DNA mismatch repair function activity of zebrafish embryos, and the application field of environmental compound DNA mismatch repair function toxicity evaluation, achieving less dosage, easy feeding, and small size Effect

Inactive Publication Date: 2012-03-28
WENZHOU MEDICAL UNIV
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Problems solved by technology

[0006] In view of this, the purpose of the present invention is to provide a simple, intuitive and effective measurement of the zebrafish embryo DNA mismatch repair functional activity analysis method and the application of this method in the evaluation of environmental compounds DNA mismatch repair functional toxicity, to overcome Traditional DNA mismatch repair functional activity analysis methods are limited to cells and cannot truly and accurately reflect the lack of DNA mismatch repair functional activity at the overall level

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  • Analysis method for activity of mismatch repair of zebra fish embryo DNA and application thereof
  • Analysis method for activity of mismatch repair of zebra fish embryo DNA and application thereof
  • Analysis method for activity of mismatch repair of zebra fish embryo DNA and application thereof

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[0036] It should be pointed out that the following specific descriptions are all exemplary, and are intended to provide further description of the present invention. Unless defined otherwise, all scientific and technical terms used herein have the same meaning as commonly understood by one skilled in the art to which this invention belongs.

[0037] The present invention will be further described below in conjunction with specific examples.

[0038] Below in conjunction with accompanying drawing for further explanation:

[0039] figure 1 Shown is the analysis method of zebrafish embryo DNA MMR activity, and the specific implementation steps are as follows.

[0040] 1. Obtaining of single-stranded DNA molecules (see flow chart figure 2 )

[0041] A. Use the helper phage M13ko7 to infect UltraMax DH5α / p111 engineering bacteria to prepare ssDNA (M13KO7 was purchased from Beijing NEB Company), where pGEM-EGFP is p111, and pGEM-(mt)EGFP is p189:

[0042] 1) One day in advance...

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Abstract

The present invention relates to an analysis method for the activity of mismatch repair of zebra fish embryo DNA, and the application of the method in toxicity evaluation of mismatch repair of environmental compound DNA. The method includes the following steps: preparing single-stranded DNA molecules with plasmids pGEM-EGFP; preparing homoduplex and heteroduplex with the single-stranded DNA molecules and linearized plasmids pGEM-EGFP and plasmids pGEM- (mt) EGFP respectively; microinjecting the homoduplex and heteroduplex to a zebra fish embryo with 1-cell stage; and quantifying the activity of mismatch repair of embryo DNA. The present invention establishes the analysis method for the activity of mismatch repair of DNA on the zebra fish embryo level for the first time, and successfully applies the method to the toxicity evaluation of mismatch repair of environmental compound DNA, which is significant for the evaluation of health risk and cancerization risk for environmental compound people. In addition, by applying the zebra fish to the evaluation of the toxicity of mismatch repair of compound DNA, the method has the advantages of simple operation, direct viewing, fastness, simple administration for tested compounds, small dosage, and the like.

Description

technical field [0001] The invention belongs to the technical field of biology, and relates to an analysis method for DNA mismatch repair function activity of zebrafish embryos, and the application of the analysis method to the toxicity evaluation of DNA mismatch repair function of environmental compounds. Background technique [0002] DNA mismatch repair (mismatch repair, MMR) system widely exists in organisms, it is a repair method after DNA replication. Its function is to repair the mismatch, insertion or deletion mutations introduced during DNA replication, and restore the DNA sequence to normal. Thus, it functions to enhance DNA replication fidelity, repair DNA mismatches, reduce genetic variation and maintain genome stability. Once the DNA mismatch repair function is weakened or lost, the body is very prone to mutations, which can induce mutation phenotypes and cause cancer. Therefore, quantitative analysis of DNA MMR functional activity is an important means for can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 董巧香黄长江陈元红陈将飞刘佳明廖军华
Owner WENZHOU MEDICAL UNIV
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