A study on the cloning and sequencing of mutation of p53 gene induced by sodium pentachlorophenate in somatic cell of zebrafish / 预防医学

Objective To explore the mutagenic effect of sodium pentachlorophenate (NaPCP) on zebrafish p53 gene coding sequence(CDS) in somatic cell.Methods The experiment was carried out using tuebingen strain of zebrafish, according to the results of acute toxicity test to determine the exposure levels in zebrafish.Zebrafish were randomly divided into blank control group and exposed groups, each containing 10 zebrafish.After exposing for 45d of NaPCP, the RNA was extracted from liver of zebra fish, and the p53 gene including a complete coding sequence of was obtained by RT-PCR.Results LC50 of NaPCP was 18.4 μg/L.Sequence analysis showed that the p53 gene CDS length of 1125bp, encoding 374 amino acids.The percent identity between the published zebrafish sequence of p53 (GI:425876786)and ours was 99.2%,with the other biological sequence of p53 existing some differences.After 45d exposure, zebrafish p53 gene of NaPCP exposure group had mutated at the concentration of 1.8 μg /L.The base substitution of GAG→AAG at codon 8,CAT→CAG at codon 148 and CAG→CAA at codon 229 were detected by PCR-directed sequencing.This may result in the Glu→Lys and His→Gln of expressed p53 protein.Conclusion NaPCP is a kind of gene mutation, which can induce the mutation of p53 gene in zebrafish somatic cells, that has the potential mutagenic risk for humans.

A study on the development toxicity and genetoxicity induced by sodium pentachlorophenate / 预防医学

Objective To investigate teratogenicity on the zebrafish embryo and genetoxicity induced by Sodium Pentachlorophenate (NaPCP),and to provide suggestions for protecting the environmental safety and human health. Methods Zebrafish embryonic development test,p53 gene in zebrafish eggs mutagenicity test,Vicia Faba Micronucleus Test and Ames test were used in the studies.Results The results of zebrafish embryonic development test showed that NaPCP increased embryonic 120 hpf teratogeny rates and 120 hpf death rates and decreased embryonic 120 hpf hatching rates in concentration of 0.5 -2.5 μL/L,with significant teratogenic effect on the embryo.The rates of embryo abnormity increased with the exposure time and concentration.Sequence analysis showed that NaPCP exposure group zebrafish p53 gene had mutated at the concentration of 0.4 μL /L.The base substitution of AAT→AAG at codon 49,ACA→ACG at codon 87 and GCG→GCA at codon 158 were detected by PCR -directed sequencing.This may result in the Asn→Lys of expressed p53 protein.As compared with control group,0.2 mg/L,1.0 mg/L dosage of NaPCP could induce an extremely significant increase in micronucleus frequency of vicia faba root tip cells and had a certain dose -effect relationship.Ames test was negative.Conclusion NaPCP could delay embryonic development and even cause embryonic lethality and induce abnormality in zebrafish.NaPCP had certain genetoxicity.