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Cloning and characterization of the HSP90 beta gene from Tanichthys albonubes Lin (Cyprinidae): effect of copper and cadmium exposure

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Abstract

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 (Cu2+) and cadmium (Cd2+) 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 Cu2+ (0, 13.50 and 27.00 μg/L) and Cd2+ (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.

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Acknowledgments

This research was supported by National Major Science and Technology Projects on Management and Control of Water Pollution in China (grants 2008ZX07211-007). We are grateful of Dr. Xichang Tan of Pearl River Research Institute, Chinese Academy of Fisheries Science for his aid in collecting experiment fish. We thank Dr. Laura Tiu of Ohio State University, USA, and Dr. Hongmei Cai of College of Resources and Environment, Huazhong Agricultural University, China, for their valuable language helps in preparing the paper.

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  1. College of Fisheries, Huazhong Agricultural University, No.1 Shizishan Street, Hongshan District, Wuhan, 430070, People’s Republic of China

    Haichao Liu, Huihui Chen, Jing Jing & Xufa Ma

  2. Ministry of Education Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of China, Wuhan, 430070, People’s Republic of China

    Haichao Liu, Huihui Chen, Jing Jing & Xufa Ma

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  1. Haichao Liu
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Correspondence to Xufa Ma.

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Liu, H., Chen, H., Jing, J. et al. Cloning and characterization of the HSP90 beta gene from Tanichthys albonubes Lin (Cyprinidae): effect of copper and cadmium exposure. Fish Physiol Biochem 38, 745–756 (2012). https://doi.org/10.1007/s10695-011-9556-2

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