Acute effect of copper and cadmium exposure on the expression of heat shock protein 70 in the Cyprinidae fish Tanichthys albonubes
Acute effect of copper and cadmium exposure on the expression of heat shock protein 70 in the Cyprinidae fish Tanichthys albonubes
复制标题
DOI：
10.1016/j.chemosphere.2013.01.014
复制
发表时间：
2013-05-01
期刊：
影响因子：
8.8
通讯作者：
Ma, Xufa
中科院分区：
文献类型：
--
作者：
Jing, Jing;Liu, Haichao;Ma, Xufa
扩增出Tanichthys albonubs热休克蛋白70(TaHSP70)的全长2398bp，包括一个1392bp的开放阅读框(ORF)，编码643个氨基酸的多肽，具有3个HSP70家族特征和EEVD的胞质基序。基因组DNA结构分析表明，TaHSP70基因在5‘端非编码区含有一个内含子。BLAST分析表明，TaHSP70基因与其他已知的HSP70基因有很高的相似性。推测的氨基酸序列也显示出高度的相似性，TaHSP70在未暴露于白纹伊蚊的不同组织中有基础表达，在肝脏中的表达水平最高。为探讨重金属暴露后TaHSP70表达的时间和剂量依赖关系，将白纹伊蚊暴露于2(-1)96h-LC_(50)(0.027 mg L-1)、4(-1)96 h-LC_(50)(0.0135 mg L-1)和2(-1)96 h-LC_(50)(2.31 mg L-1)、4(-1)96 h-LC_(50)(1.15 mg L-1)96h，用实时定量聚合酶链式反应和Western blotting分析对照细胞热休克蛋白70的表达。结果表明，肝脏对重金属胁迫的反应存在剂量依赖性表达模式和暴露时间效应。有趣的是，TaHSP70基因的表达在转录和翻译水平之间并没有显示出一致的变化。综上所述，观察到的TaHSP70的表达动态为了解重金属胁迫、热休克蛋白活性以及监测白毛虫培养环境中慢性应激源的潜在途径提供了重要的见解。(C)2013爱思唯尔有限公司。保留所有权利。
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) 96 h-LC50 (0.027 mg L-1), 4(-1) 96 h-LC50 (0.0135 mg L-1) of copper and 2(-1) 96 h-LC50 (2.31 mg L-1), 4(-1) 96 h-LC50 (1.15 mg L-1) of cadmium for 96 h. 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. (C) 2013 Elsevier Ltd. All rights reserved.