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为研究稀有逗鲫(Gobiocypris rarus)HSP70用于淡水毒理学和风险评估方面的可行性,通过cDNA末端快速扩增的方法首次从稀有逗鲫肝脏中成功克隆出HSP70基因的cDNA全长,命名为GrHSP70。NCBI比对分析结果显示,GrHSP70核苷酸序列与其他鱼类的HSP70基因的核苷酸序列相似性极高,相应的氨基酸序列同源性也高(大于95%)。通过实时定量PCR获得未经污染物暴露的稀有逗鲫幼鱼体内13种组织中GrHSP70的分布以及PCP暴露后肝脏中该基因的表达图谱,结果表明GrHSP70在稀有逗鲫体内的表达呈现出组织依赖性,在性腺组织中的表达最高。此外,在不同时间﹑不同浓度的PCP暴露下稀有逗鲫肝脏中GrHSP70的表达模式呈现出显著的时间-剂量依赖效应。总之,GrHSP70可作为一种非常敏感的生物标志物,适用于中国淡水环境污染的毒理学研究及风险评估。
In order to study the feasibility of Gobiocypris rarus HSP70 for freshwater toxicology and risk assessment, the full-length cDNA of HSP70 gene was successfully cloned from the liver of Ragginous Carassius auratus by rapid amplification of cDNA ends and named as GrHSP70. The results of NCBI alignment showed that the nucleotide sequence of GrHSP70 was highly similar to that of other fish HSP70 genes, and the corresponding amino acid sequence homology was high (more than 95%). The distribution of GrHSP70 in 13 tissues and the expression pattern of this gene in liver after exposure to PCP were obtained by real-time PCR. The results showed that the expression of GrHSP70 in tissues of RAT Sex, the highest expression in gonadal tissue. In addition, the expression pattern of GrHSP70 in the liver of the rare funny carp exposed to different concentrations of PCP at different times showed a significant time-dose-dependent effect. In conclusion, GrHSP70 can be used as a very sensitive biomarker for toxicology research and risk assessment of freshwater environmental pollution in China.