Studies On Exon 8 Mutation Of P53 Gene Of Rats' Spermary Caused By ZEN By PCR-SSCP

Zearlenone, i.e.F-2 toxin, is an epiphyte toxin which has estrogen action produced by Gibberlla zeae, F. graminearum, Fudarium tricinctum and so on. It mainly empoison feed and cropper, such as wheat, oat, corn and so on. It can arouse uterine portliness and abortion of female livestock, female symptom of boar which have an enormously impact on the breed aquatics industry. Therefore, it is very necessary to probe into the influence of procreation caused by mutation of gene, and how to reduce the effect of ZEN on breed and growth of animals.Firstly, by take different dosage of ZEN into rats, utilize reagent of Zhongkekairui Co., and distill the DNA of different rates, the result shows that the pure of DNA is high, the content is about 170ug/ml, which is fit for the template of PCR.Secondly, by use PCR premier and PCR pre-reaction reagent box, carry through the PCR reaction, identify the pure of outcome by 0.8% gelatin electrophoresis, the result shows that the outcome of PCR is exclusive and about 200bp Mol. wt. ,which is very close to full size, therefore ,is the Exon 8 Mutation of p53 Gene.Thirdly, by SSCP analysis of the PCR outcome, the result shows that there are discrepancies in DNA strip of different groups. It means that ZEN had an effect on framework of the Exon 8 Mutation of P53 Gene. And then, the mutation of gene was discovered by measuring the sequence of gene. The mutation occurs in 5'end ,but the sequence of the next 20 bp is similar. There is only 1 bp between comparison of solvent group and testing group of 100μg/kg wt ZEN. The frequency of mutation is about 3% between other testing group and comparison of group.