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Jing J.,Huazhong Agricultural University | Jing J.,Ministry of Education Key Laboratory of Agricultural Animal Genetics | Liu H.,Huazhong Agricultural University | Liu H.,Ministry of Education Key Laboratory of Agricultural Animal Genetics | And 8 more authors.
Chemosphere | Year: 2013

A full sequence of TaHSP70 (heat shock protein 70 of Tanichthys albonubes) was amplified which was 2398. bp, including an open reading frame (ORF) of 1392. bp encoding a polypeptide of 643 amino acids with all three HSP70 family signatures and cytosolic motif of EEVD. Genomic DNA structure analysis revealed that the TaHSP70 gene contained one intron in 5'UTR. BLAST analysis revealed that the TaHSP70 gene shared high similarity with other known HSP70 genes. The alignment of inferred amino acid sequences also showed high degrees of similarity among the homologues. There was a basal mRNA expression of TaHSP70 in the different tissues from the non-exposed T. albonubes and the highest expression level in the liver. To investigated the time- and dose-dependent relationship of the expression of TaHSP70 following exposure to heavy metals, T. albonubes were exposed to 2-1 96h-LC50 (0.027mgL-1), 4-1 96h-LC50 (0.0135mgL-1) of copper and 2-1 96h-LC50 (2.31mgL-1), 4-1 96h-LC50 (1.15mgL-1) of cadmium for 96h. Hsp70 expression relative to the control was analyzed by Quantitative real-time PCR and Western blotting. The results indicated that there were a dose-dependent expression pattern and an exposure time effect in the liver responded to heavy metal stress. Interestingly, TaHSP70 gene expressions did not show consistent changes between transcription and translation levels. Taken together, the dynamics of TaHSP70 expression observed provided important insight into heavy metal stress, heat shock protein activity, and the potential ways to monitor the chronic stressors in T. albonubes culture environments. © 2013 Elsevier Ltd.


Liu H.,Huazhong Agricultural University | Liu H.,Ministry of Education Key Laboratory of Agricultural Animal Genetics | Chen H.,Huazhong Agricultural University | Chen H.,Ministry of Education Key Laboratory of Agricultural Animal Genetics | And 4 more authors.
Fish Physiology and Biochemistry | Year: 2012

Heat shock protein 90 (HSP90) is a highly conserved molecular chaperone important in the maturation of a broad spectrum of proteins. In order to evaluate the effect of copper (Cu 2+) and cadmium (Cd 2+) on the expression of HSP90 from Tanichthys albonubes (designated TaHSP90), the full-length complementary DNA (cDNA) of TaHSP90 was cloned using reverse transcription PCR and rapid amplification of cDNA ends (RACE) techniques. A 2,687-bp sequence was sequenced and consisted of an open reading frame (ORF) of 2,181 bp encoding a polypeptide of 727 amino acids with five HSP90 family signatures. Homologous analysis revealed that TaHSP90 gene shared high similarity with other known HSP90 genes and belonged to HSP90β subtype. Fluorescent real-time quantitative PCR was used to examine the expression pattern of TaHSP90β mRNA in different tissues (liver, muscle, gill, fin, eye, ovary, intestine and brain), and the result indicated that TaHSP90β was widely expressed in all examined tissues at different levels. Sensitivity of TaHSP90β to copper and cadmium was examined by exposing fish to different concentrations of Cu 2+ (0, 13. 50 and 27. 00 μg/L) and Cd 2+ (0, 1. 15, 2. 31 mg/L) for 24, 48, 72 and 96 h, respectively. The copper treatment induced TaHSP90β expression slight increase only at 24 and 48 h, while cadmium treatment caused slight down-regulation of TaHSP90β only 72 and 96 h. Our data suggest that the cloning and expression analysis of T. albonubes HSP90β gene provided useful molecular information of T. albonubes responses in stress conditions and potential ways to monitor the chronic stressors in T. albonubes culture environments. © 2011 Springer Science+Business Media B.V.

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