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

Abstract

Heat shock protein (HSP) 70, as a molecular chaperone and extensively studied protein in environmental toxicology, plays a key role in membrane translocation, protein degradation, and regulatory process. However, as toxicology and risk assessment of freshwater, characterizations of HSP 70 in rare minnow, Gobiocypris rarus , remain unknown. In this paper, we described the full-length cDNA of HSP 70, designated as GrHSP 70, by cloning through rapid-amplification of cDNA ends (RACE) technique from the liver of Gobiocypris rarus for the first time. The GrHSP 70 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 GrHSP 70 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 GrHSP 70 is tissue-dependent, with the highest expression in the gonad tissue. Moreover, GrHSP 70 is significantly up-regulated in liver by a concentration/time-depen-dent manner after PCP exposure. In summary, our results suggest that GrHSP 70 may serve as a very sensi-tive biomarker for a toxicology and risk assessment of freshwater pollution in China.