条斑紫菜HSP90基因的克隆与表达分析  

Molecular cloning and expression analysis of HSP90 gene from Porphyra yezoensis Ueda (Bangiales,Rhodophyta)

作　　者：周向红[1,2] 李信书[1,2] 王萍[2] 阎斌伦[1] 滕亚娟[2] 易乐飞[1,2] 

机构地区：[1]淮海工学院江苏省海洋生物技术重点实验室,江苏连云港222005 [2]淮海工学院海洋学院,江苏连云港222005

出　　处：《水产学报》2010年第12期1844-1852,共9页Journal of Fisheries of China

基　　金：国家"十一五"科技支撑计划重大项目(2006BAD09A01);江苏省海洋生物技术重点实验室开放课题(2008HS004)

摘　　要：为了研究HSP90在条斑紫菜胁迫耐受中的作用,克隆了条斑紫菜HSP90基因(命名为PyHSP90),采用定量RT-PCR研究了其表达规律。PyHSP90基因包含一个长2274nt的完整开放阅读框,编码757个氨基酸。PyHSP90蛋白定位于细胞质中,具有HSP90蛋白家族的特征序列和保守结构域,与多种生物的HSP90具有较高的序列一致性。在进化树上条斑紫菜等红藻的HSP90与隐藻的亲缘关系最近,与绿藻和陆生植物亲缘关系较远。低温和高温胁迫都能显著性地诱导条斑紫菜叶状体PyHSP90基因的表达,而且表达量与胁迫程度成正相关;低盐胁迫下PyHSP90基因表达上调;而失水胁迫下PyHSP90基因表达下调。The 90-ku heat shock proteins（HSP90s） are a group of evolutionarily highly conservative molecular chaperones and essential for viability in eukaryotes. The basic functions of HSP90s are of assisting protein folding,protein degradation and protein trafficking,and they also play an important role in signal transduction networks,cell cycle control and morphological evolution. Although HSP90s are constitutively expressed in most organisms,their expression is up-regulated in response to stresses such as cold,heat,salt stress,heavy metals,phytohormones,light and dark transitions. Porphyra yezoensis Ueda,one of intertidal macroalgae,periodically experiences immersion and emersion due to the tidal cycles. When P. yezoensis thalli are exposed to air,they experience a variety of abiotic stresses such as nutrient limitation,high light,ultraviolet ray,high and low temperature,desiccation and osmotic stress. In order to investigate the contribution of HSP90 to abiotic stresses tolerance in P. yezoensis thalli,molecular cloning and expression analysis of HSP90 gene（designated as PyHSP90） from P. yezoensis were performed. PyHSP90 gene was obtained with computer assisted cloning,and characterized using multiple bioinformatic programs. The relative mRNA expression levels of PyHSP90 gene under stresses were investigated in P. yezoensis thalli using real-time RT-PCR. The PyHSP90 cDNA contained a 2 274 nt of continuous complete open reading frame encoding a polypeptide of 757 amino acids with a calculated molecular mass of 86 ku. Analytic results of PSORT,TargetP and phylogenetic tree showed that PyHSP90 localized to cytoplasm. PyHSP90 contained conserved signature motifs and domains of HSP90 family. PyHSP90 shared high amino acid sequence identity with HSP90s from other organisms. Sequence comparison of PyHSP90 revealed 26. 0%-39. 4% and 61. 4%-71. 8% identity with prokaryotic and eukaryotic HSP90s respectively. In the phylogenetic tree red algae HSP90s,including PyHSP90,clustered with Cryptophyta HSP90s firstly,and then cl

关 键 词：条斑紫菜 热应激蛋白90 克隆 表达 定量PCR 

分 类 号：S917[农业科学—水产科学] Q781[生物学—分子生物学]