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。
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 5rUTR 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.
出处
《海洋与湖沼》
CAS
CSCD
北大核心
2012年第6期1156-1162,共7页
Oceanologia Et Limnologia Sinica
基金
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
关键词
浙江枝吻纽虫
热休克蛋白70
实时荧光定量PCR
表达
Dendrorhynchus zhejiangensis, Heat shock protein 70 （HSP70）, Real-time fluorescent quantitative PCR, Expression
