坛紫菜两种Hsp90基因的克隆及表达特征分析  

Cloning and expression pattern analysis of two heat shock protein (Hsp90) genes from Pyropia haitanensis

作　　者：代真真[1] 李兵[1] 徐燕[1] 纪德华[1] 陈昌生[1] 谢潮添[1] 

机构地区：[1]集美大学水产学院,福建厦门361021

出　　处：《水产学报》2014年第3期340-349,共10页Journal of Fisheries of China

基　　金：国家"八六三"高技术研究发展计划(2012AA10A411);国家自然科学基金项目(41176151;41276177);海洋公益性行业科研专项(201105008;201105023);福建省科技重大专项资助项目(2011NZ0001)

摘　　要：以坛紫菜转录组测序获得的unigene序列为基础,采用RACE(rapid amplification of cDNA ends)技术克隆获得了坛紫菜的两条Hsp90基因序列:PhHsp90-1和PhHsp90-2。序列分析结果表明,PhHsp90-1序列全长2 572 bp,包含一个2 427 bp的开放阅读框,所编码的多肽包含809个氨基酸,分子量为90.2 ku,等电点为4.79,属于Hsp90内质网亚家族(登录号:KF732651);PhHsp90-2序列全长2 510 bp,包含一个2 280 bp的开放阅读框,所编码的多肽包含760个氨基酸,分子量为86.2 ku,等电点为4.81,属于Hsp90细胞质亚家族(登录号:KF732652)。基因表达水平的定量分析结果表明,高温和失水胁迫对两条PhHsp90基因的表达水平均有显著影响,但在表达模式上存在明显差异。高温胁迫不同时间水平下,两条PhHsp90基因的表达均表现为先上调后下调的趋势;而在失水胁迫下,当失水率小于60%时,两条基因的表达水平均没有发生显著变化,但当失水率大于60%时,两条基因的表达水平均显著上调,说明两条PhHsp90基因均在应答高温胁迫和高度失水胁迫中发挥着重要作用。Heat shock protein（ Hsp90）, representing an important molecular chaperone in eukaryotic cells, plays particularly important roles in a variety of stress responses, development and signal transduction of plants. In this study,based on unigene sequences which were obtained from whole transcriptome sequencing of P. haitanensis, two full-length PhHsp90 genes were obtained by rapid amplification of cDNA ends （RACE） ,and named PhHsp90-1 and PhHsp90-2. The full-length cDNA of the PhHsp90-1 gene comprised 2 572 nucleotides and contained an open reading frame of 2 427 bp （ GenBank accession： KF732652）, encoding a protein of 809 amino acid residues with the predicted molecular weight of 90. 2 kDa and theoretical isoelectric point of 4.79; and the full-length cDNA of the PhHsp90-2 gene comprised 2 510 nucleotides and contained an open reading frame of 2280bp （ GenBank accession： KF732651 ）, encoding a protein of 760 amino acid residues with the predicted molecular weight of 86. 2 kDa and theoretical isoelectric point of 4.81. On the basis of conserved motifs and phylogenetic tree analysis, PhHsp90-1 belongs to the endoplasmic reticulum subfamily of Hsp90 and PhHsp90-2 belongs to the cytoplasmic subfamily of Hsp90. The expressions of the two PhHsp90 genes, as measured by real-time quantitative PCR, were significantly induced by high-temperature stress and desiccation stress ,but had different expression patterns. Under high-temperature stress, the expression levels of the two PhHsp90 genes all significantly increased first and then decreased. However, during desiccation, the expression levels of PhHsp90-1 and PhHsp90-2 were significantly increased only when the water loss was 〉 60%. These results suggested that the two PhHsp90s play important roles in the response to high-temperature stress and extreme desiccation stress.

关 键 词：坛紫菜 热激蛋白90 RACE 荧光定量PCR 

分 类 号：Q785[生物学—分子生物学] S917[农业科学—水产科学]