引用本文:	路雅丽,李成华,苏秀榕,李太武,朱琳,张鹏,李晔.浙江枝吻纽虫(Dendrorhynchus zhejiangensis) 热休克蛋白70基因克隆与表达特征研究.海洋与湖沼,2012,43(6):1156-1162.

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*浙江枝吻纽虫(Dendrorhynchus zhejiangensis) 热休克蛋白70基因克隆与表达特征研究*
路雅丽¹, 李成华¹, 苏秀榕¹, 李太武^1,2, 朱琳¹, 张鹏¹, 李晔¹
1.宁波大学海洋学院;2.宁波城市职业技术学院

摘要:

采用同源克隆和cDNA 末端快速扩增技术(RACE)克隆了浙江枝吻纽虫热休克蛋白70(DzHSP70)基因全长cDNA序列, 并利用荧光定量PCR方法, 分析了该基因在重金属胁迫下的表达特征。结果表明, DzHSP70cDNA 全长2827bp, 包括294bp的5’UTR、628bp的3’UTR和编码634个氨基酸残基的1905bp 的开放阅读框。HSP70家族的3 个签名序列以及细胞质特异性调控基序EEVD等特征元件在DzHSP70中高度保守。重金属离子Fe³⁺、Pb²⁺和Cd²⁺均可上调该基因的表达, 其中Pb²⁺的毒理效应最强。研究结果表明DzHSP70可能参与了机体对于重金属的解毒过程。

关键词: 浙江枝吻纽虫, 热休克蛋白70, 实时荧光定量PCR, 表达

DOI：10.11693/hyhz201206018018

分类号:

基金项目:浙江省自然科学基金资助项目, Y3100480 号; 浙江省教育厅(理), Y200907951号; 国家自然科学基金项目, 31272637 号。

附件

CLONING AND EXPRESSION ANALYSIS OF HEAT SHOCK PROTEIN 70 FROM DENDRORHYNCHUS ZHEJIANGENSIS

LU Ya-Li¹, LI Cheng-Hua¹, SU Xiu-Rong¹, LI Tai-Wu^1,2, ZHU Lin¹, ZHANG Peng¹, LI Ye¹

1.School of Marine Sciences, Ningbo University;2.Ningbo City College of Vocational Technology

Abstract:

With the approaches of homology cloning, RACE, the full-length HSP70 cDNA of Dendrorhynchus zhejiangensis was identified and characterized (denoted as DzHSP70) in the present study. Meanwhile, fluorescent real-time PCR was used to analyze varying degrees of induction of DzHSP70 mRNA expression in D. zhejiangensis exposed to heavy metals (Fe³⁺, Pb²⁺ and Cd²⁺). The results indicated the cDNA of DzHSP70 was of 2827bp, consisting of a 5′UTR of 294bp, a 3′UTR of 628bp, and a complete open reading frame of 1905bp encoding a polypeptide with 566 amino acid residues. The conserved motifs for HSP70 protein family including three signature sequences and a cytoplasm characteristic motif of EEVD were totally found in the deduced amino acid of DzHSP70. Three heavy metals Fe³⁺, Pb²⁺ and Cd²⁺ all could be induced the expression of DzHSP70 at mRNA level, where Pb²⁺ displayed more toxic effect on worm than those of Fe³⁺ and Cd²⁺. All our results indicated that DzHSP70 might be involved into mediating the process of heavy metals detoxification in worm.

Key words: Dendrorhynchus zhejiangensis, Heat shock protein 70 (HSP70), Real-time fluorescent quantitative PCR, Expression