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

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...

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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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