Molecular cloning, characterization, and expression analysis of a cytosolic HSP90 gene from Haematococcus pluvialis
文献类型：期刊论文
| 作者 | Zhang, Lei1,2; Fan, Yinmao1,3; Shi, Fei1,2; Qin, Song1 ; Liu, Bing1
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| 刊名 | JOURNAL OF APPLIED PHYCOLOGY
 |
| 出版日期 | 2012-12-01 |
| 卷号 | 24期号:6页码:1601-1612 |
| 关键词 | Haematococcus pluvialis Heat shock protein 90 cDNA cloning qRT-PCR mRNA expression |
| ISSN号 | 0921-8971 |
| 产权排序 | [Zhang, Lei; Fan, Yinmao; Shi, Fei; Qin, Song; Liu, Bing] Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China; [Zhang, Lei; Shi, Fei] Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China; [Fan, Yinmao] Ningbo Univ, Fac Life Sci & Biotechnol, Ningbo 315211, Zhejiang, Peoples R China |
| 通讯作者 | Qin, S (reprint author), Chinese Acad Sci, Yantai Inst Coastal Zone Res, 17 Chunhui Rd, Yantai 264003, Peoples R China. |
| 中文摘要 | Heat shock protein 90 (HSP90) is a highly conserved molecular chaperone that plays key roles in the folding, maintenance of structural integrity, and regulation of a subset of cytosolic proteins. In this study, the cDNA of Haematococcus pluvialis HSP90 (designated HpHSP90) was cloned by the combination of homology cloning and rapid amplification of cDNA ends approaches. The full-length cDNA of HpHSP90 was of 2,606 bp, including an open reading frame of 2,109 bp encoding a polypeptide of 702 amino acids with predicted molecular weight of 80.14 kDa and theoretical isoelectric point of 5.07. BLAST analysis revealed that HpHSP90 shared high similarity with other known HSP90s, and the five conserved amino acid blocks defined as HSP90 protein family signatures were also identified in HpHSP90, which indicated that HpHSP90 should be a cytosolic member of the HSP90 family. Under different stress conditions, messenger RNA (mRNA) expression levels of HpHSP90 were quantified by quantitative RT-PCR. To H. pluvialis kept at different temperatures for 1 h, maximum HpHSP90 expression was observed in the range 5 to 10A degrees C and 35 to 40A degrees C and the expression level of HpHSP90 at 40A degrees C was the highest (threefold compared with that at 25A degrees C). In H. pluvialis kept at 35A degrees C for different times, the mRNA expression level of HpHSP90 reached a maximum level after 7 h and then dropped progressively. The results indicate that HpHSP90 responded to cold and heat stresses with a temperature-dependent expression pattern as well as exposure time effect and could be used as a molecular biomarker in adverse stress environment. |
| 英文摘要 | Heat shock protein 90 (HSP90) is a highly conserved molecular chaperone that plays key roles in the folding, maintenance of structural integrity, and regulation of a subset of cytosolic proteins. In this study, the cDNA of Haematococcus pluvialis HSP90 (designated HpHSP90) was cloned by the combination of homology cloning and rapid amplification of cDNA ends approaches. The full-length cDNA of HpHSP90 was of 2,606 bp, including an open reading frame of 2,109 bp encoding a polypeptide of 702 amino acids with predicted molecular weight of 80.14 kDa and theoretical isoelectric point of 5.07. BLAST analysis revealed that HpHSP90 shared high similarity with other known HSP90s, and the five conserved amino acid blocks defined as HSP90 protein family signatures were also identified in HpHSP90, which indicated that HpHSP90 should be a cytosolic member of the HSP90 family. Under different stress conditions, messenger RNA (mRNA) expression levels of HpHSP90 were quantified by quantitative RT-PCR. To H. pluvialis kept at different temperatures for 1 h, maximum HpHSP90 expression was observed in the range 5 to 10A degrees C and 35 to 40A degrees C and the expression level of HpHSP90 at 40A degrees C was the highest (threefold compared with that at 25A degrees C). In H. pluvialis kept at 35A degrees C for different times, the mRNA expression level of HpHSP90 reached a maximum level after 7 h and then dropped progressively. The results indicate that HpHSP90 responded to cold and heat stresses with a temperature-dependent expression pattern as well as exposure time effect and could be used as a molecular biomarker in adverse stress environment. |
| 学科主题 | Biotechnology & Applied Microbiology; Marine & Freshwater Biology |
| 研究领域[WOS] | Biotechnology & Applied Microbiology ; Marine & Freshwater Biology |
| 关键词[WOS] | HEAT-SHOCK-PROTEIN ; CAROTENOID BIOSYNTHESIS GENES ; PHYLOGENETIC ANALYSIS ; SEQUENCE ALIGNMENT ; ESCHERICHIA-COLI ; THERMAL-STRESS ; CDNA ; INDUCTION ; CADMIUM ; CELLS |
| 收录类别 | SCI |
| 资助信息 | CAS; Outstanding young scholar fellowship of Shandong Province [JQ200914]; National Natural Science Foundation of China [31000037]; Knowledge Innovation Program of the Chinese Academy of Sciences [KSCX2-YW-G-073, KZCX2-YW-216, KZCX2-YW-209] |
| 原文出处 | http://dx.doi.org/s10811-012-9821-5 |
| 语种 | 英语 |
| WOS记录号 | WOS:000310232900027 |
| 公开日期 | 2013-03-08 |
| 源URL | [http://ir.yic.ac.cn/handle/133337/6123]  |
| 专题 | 烟台海岸带研究所_海岸带生物学与生物资源利用所重点实验室 |
| 作者单位 | 1.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China 3.Ningbo Univ, Fac Life Sci & Biotechnol, Ningbo 315211, Zhejiang, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, Lei,Fan, Yinmao,Shi, Fei,et al. Molecular cloning, characterization, and expression analysis of a cytosolic HSP90 gene from Haematococcus pluvialis[J]. JOURNAL OF APPLIED PHYCOLOGY,2012,24(6):1601-1612. |
| APA | Zhang, Lei,Fan, Yinmao,Shi, Fei,Qin, Song,&Liu, Bing.(2012).Molecular cloning, characterization, and expression analysis of a cytosolic HSP90 gene from Haematococcus pluvialis.JOURNAL OF APPLIED PHYCOLOGY,24(6),1601-1612. |
| MLA | Zhang, Lei,et al."Molecular cloning, characterization, and expression analysis of a cytosolic HSP90 gene from Haematococcus pluvialis".JOURNAL OF APPLIED PHYCOLOGY 24.6(2012):1601-1612. |
入库方式： OAI收割
来源：烟台海岸带研究所
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