【摘要】 将斑马鱼经50μg/L五氯酚(PCP)体内暴露染毒10d,提取肝脏基因组DNA,通过巢式PCR(nestedPCR)扩增包含斑马鱼p53基因外显子2,3,4及其间内含子2,3的片段,扩增产物分别用限制性内切酶NcoI、SacI、ScaI、BclI、NsiI、RsaI进行限制性片段长度多态性(RFLP)分析,以检测目标片段在酶切位点上点突变的发生.检测了20条斑马鱼上述6个酶切位点上的共680个碱基,但均未检测到点突变.结果表明,如果PCP能够诱导斑马鱼p53基因点突变,则其碱基突变率小于1/680.更多还原

【Abstract】 The hepatic genomic DNA of zebrafish was extracted after contamination exposure to 50μg/L pentachlo-rophenol (PCP) for 10d in vivo. A fragment containing exon2, 3, 4 and intron2, 3 of zebrafish p53 gene was amplified through nested PCR-RFLP analysis of the amplified products was carried out using restriction enzymes Nco I, Sac I, Sca I, Bcl I, Nsi I and Rsa I respectively, to detect the occurrence of point mutation of the objective fragment on the enzyme site. 680bp of the above 6 enzyme sites of 20 zebrafishes were detected, but no point mutation was detected. If p53 gene point mutation could be induced by PCP in zebrafish, the mutation rate of its bp was less than 1/680.更多还原