稀有鲫热激蛋白70的分子克隆及表达特征分析（英文）  

MOLECULAR CLONING AND EXPRESSION CHARACTERIZATION OF HEAT SHOCK PROTEIN 70 FROM GOBIOCYPRIS RARUS

作　　者：黄书婷[1] 刘秋萍[1] 邓川[1] 和野 熊力[1] 牛纯青[1] 陈韵[1] 刘堰[1] 

机构地区：[1]西南大学生命科学学院,淡水鱼类资源与生殖发育教育部重点实验室,三峡库区生态环境教育部重点实验室,水产科学重庆市市级重点实验室,重庆400715

出　　处：《水生生物学报》2017年第1期18-25,共8页Acta Hydrobiologica Sinica

基　　金：Supported by the National Program on Key Basic Research Project(973 Program;2012CB723205);the Financial Support Received from the National Natural Science Foundation of China(21147002)

摘　　要：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。Heat shock protein （HSP） 70, as a molecular chaperone and extensively studied protein in environ-mental toxicology, plays a key role in membrane translocation, protein degradation, and regulatory process. However, as toxicology and risk assessment of freshwater, characterizations of HSP70 in rare minnow, Gobio-cypris rarus, remain unknown. In this paper, we described the full-length cDNA of HSP70, designated as GrHSP70, by cloning through rapid-amplification of cDNA ends （RACE） technique from the liver of Gobio-cypris rarus for the first time. The GrHSP70 shares high nucleotide sequence homolog, corresponding to amino acid sequence homology （〉95%） with other fish species by NCBI blast analysis. Through quantitative real-time PCR （RT-PCR）, we obtained the distribution of GrHSP70 in thirteen tissues of the untreated Gobio-cypris rarus juveniles and its expression profile in liver after pentachlorophenol （PCP） exposure. The re-sults indicated that mRNA transcription level of GrHSP70 is tissue-dependent, with the highest expression in the gonad tissue. Moreover, GrHSP70 is significantly up-regulated in liver by a concentration/time-depen-dent manner after PCP exposure. In summary, our results suggest that GrHSP70 may serve as a very sensi-tive biomarker for a toxicology and risk assessment of freshwater pollution in China.

关 键 词：稀有逗鲫 热激蛋白70 CDNA末端快速扩增 五氯酚 生物标志物 HSP70 

分 类 号：Q344.1[生物学—遗传学]