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双酚A对斑马鱼急性毒性及P53基因诱变性影响

P53 gene mutation and acute toxicity in zebrafish exposed to BPA

  • 摘要: 目的 研究双酚A(bisphenol A,BPA)对斑马鱼P53基因诱变性.方法 根据斑马鱼急性暴露试验结果,确定斑马鱼暴露浓度.试验设对照组与双酚A暴露组,连续染毒30 d,从斑马鱼肝脏组织中提取RNA,通过逆转录-聚合酶链反应(RT-PCR)方法扩增得到有完整编码区序列的P53基因,并进行测序分析.结果 序列分析表明,斑马鱼P53基因编码区序列长1 122 bp,编码373个氨基酸,与网上已经登陆斑马鱼P53序列有99.9%相似系数,而与其他生物P53基因序列存在一定差异;在0.3 mg/L双酚A染毒30 d后,双酚A暴露组斑马鱼P53基因序列发生了突变;直接测序证明,分别在861、35、311位密码子发生了ACA→ACG、GAG→GGG、AGG→CGG转变,可使其编码P53蛋白氨基酸发生谷氨酸→甘氨酸变异.结论 双酚A对斑马鱼P53基因有诱变性.

     

    Abstract: Objective To study the mutagenic effect of bisphenol A(BPA) on P53 gene in zebrafish.Methods The exposure levels of BPA were determined according to the results of acute toxicity test.Twenty zebrafishes were randomly divided into blank control and exposure group(10 fish in each group).After exposure to BPA for 30 days,the RNA was extracted from liver tissue of the zebrafish and the P53 gene,including a complete coding sequence,was obtained by reverse transcription-PCR(RT-PCR).Results Sequence analyses showed that the length coding sequences(CDS) of the P53 gene is 1 122bp,encoding 373 amino acids and the identical percentage between the published zebrafish sequence and the P53 gene identified is 99.9%,with some differences form P53 of other biotic.After 30 days' exposure,the P53 gene of the exposure group mutated at the concentration of 0.3mg/L.The base substitution of ACA→ACG at codon 86,GAG→GGG at codon 135,AGG→CGG at codon 311 were detected by PCR-directed sequencing.This may result in the Glu→Gly of expressed P53 protein.Conclusion The results show that BPA could induce mutation in P53 gene.Under the experimental conditions,the lowest obsvered effect level(LOFL) of BAA is 0.3mg/L.

     

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