Molecular cloning and expression analysis of a cytosolic Hsp70 gene from Ulva pertusa (Ulvophyceae, Chlorophyta)

doi:10.1007/s10811-010-9561-3

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	Molecular cloning and expression analysis of a cytosolic Hsp70 gene from Ulva pertusa (Ulvophyceae, Chlorophyta)
	Fu, Wandong 1,2; Shuai, Li 3; Yao, Jianting 1; Zheng, Bin 2; Zhong, Mingjie 2; Duan, Delin 1
	2011-08-01
发表期刊	JOURNAL OF APPLIED PHYCOLOGY
ISSN	0921-8971
卷号	23 期号:4 页码:681-690
文章类型	Article
摘要	In this study, homologous cloning coupled with the rapid amplification of cDNA ends was used to clone a full-length cytosolic heat shock protein 70 of Ulva pertusa (designated as UPHsp70). Bioinformatics was used to analyze structural features, homologous relationship, and phylogenetic position of UPHsp70. The full length of UPHsp70 cDNA was 2,283 bp, with a 5' untranslated region of 65 bp, a 3' untranslated region of 247 bp, and an open reading frame of 1,971 bp encoding a polypeptide of 656 amino acids with an estimated molecular weight of 71.13 kDa and an estimated isoelectric point of 5.04. The UPHsp70 had four degenerate repeats of tetrapeptide GGMP and three typical Hsp70 signature motifs. The specific C-terminus amino acid sequence of cytosolic UPHsp70 was EEVD, and the conservation of Hsp70s of N-terminus was higher than that of C-terminus. The homology between UPHsp70 and Hsp70s of other known algae and land plants was more than 70%. Under different stress conditions, mRNA expression levels of UPHsp70 were quantified by quantitative reverse transcriptase-polymerase chain reaction. When U. pertusa samples were kept in different temperatures (5-40A degrees C) for 1 h, the expression level of UPHsp70 at 5A degrees C, 35A degrees C, or 40A degrees C was over onefold higher than that at 25A degrees C. When U. pertusa samples were kept at 30A degrees C for different times (0-12 h), the mRNA expression level of UPHsp70 had a trend of rise first then fall. The expression level of UPHsp70 reached maximum level after 5 h. When U. pertusa samples were kept in different salt concentrations (0-45aEuro degrees) for 2 h, the expression level of UPHsp70 at 0aEuro degrees or 5aEuro degrees salt concentration was twofold higher than that at 30aEuro degrees for 2 h. The expression levels of UPHsp70 at 30aEuro degrees, 35aEuro degrees, and 40aEuro degrees were low and had no difference (P < 0.05). When U. pertusa samples were kept at ultraviolet irradiation or desiccated for different times (0-4 h), the mRNA expression level of UPHsp70 reached maximum level after 3.0 h; after that, it was maintained at high level.; In this study, homologous cloning coupled with the rapid amplification of cDNA ends was used to clone a full-length cytosolic heat shock protein 70 of Ulva pertusa (designated as UPHsp70). Bioinformatics was used to analyze structural features, homologous relationship, and phylogenetic position of UPHsp70. The full length of UPHsp70 cDNA was 2,283 bp, with a 5' untranslated region of 65 bp, a 3' untranslated region of 247 bp, and an open reading frame of 1,971 bp encoding a polypeptide of 656 amino acids with an estimated molecular weight of 71.13 kDa and an estimated isoelectric point of 5.04. The UPHsp70 had four degenerate repeats of tetrapeptide GGMP and three typical Hsp70 signature motifs. The specific C-terminus amino acid sequence of cytosolic UPHsp70 was EEVD, and the conservation of Hsp70s of N-terminus was higher than that of C-terminus. The homology between UPHsp70 and Hsp70s of other known algae and land plants was more than 70%. Under different stress conditions, mRNA expression levels of UPHsp70 were quantified by quantitative reverse transcriptase-polymerase chain reaction. When U. pertusa samples were kept in different temperatures (5-40A degrees C) for 1 h, the expression level of UPHsp70 at 5A degrees C, 35A degrees C, or 40A degrees C was over onefold higher than that at 25A degrees C. When U. pertusa samples were kept at 30A degrees C for different times (0-12 h), the mRNA expression level of UPHsp70 had a trend of rise first then fall. The expression level of UPHsp70 reached maximum level after 5 h. When U. pertusa samples were kept in different salt concentrations (0-45aEuro degrees) for 2 h, the expression level of UPHsp70 at 0aEuro degrees or 5aEuro degrees salt concentration was twofold higher than that at 30aEuro degrees for 2 h. The expression levels of UPHsp70 at 30aEuro degrees, 35aEuro degrees, and 40aEuro degrees were low and had no difference (P < 0.05). When U. pertusa samples were kept at ultraviolet irradiation or desiccated for different times (0-4 h), the mRNA expression level of UPHsp70 reached maximum level after 3.0 h; after that, it was maintained at high level.
关键词	Ulva Pertusa Heat Shock Protein 70 Molecular Cloning Mrna Expression
学科领域	Biotechnology & Applied Microbiology ; Marine & Freshwater Biology
DOI	10.1007/s10811-010-9561-3
URL	查看原文
收录类别	SCI
语种	英语
WOS记录号	WOS:000293163500005
引用统计	被引频次：11[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/11901
专题	实验海洋生物学重点实验室
作者单位	1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China 2.Zhejiang Marine Dev Res Inst, Zhoushan 316100, Peoples R China 3.Qingdao Univ, Coll Chem Chem Engn & Environm Sci, Qingdao 266071, Peoples R China
第一作者单位	中国科学院海洋研究所
推荐引用方式 GB/T 7714	Fu, Wandong,Shuai, Li,Yao, Jianting,et al. Molecular cloning and expression analysis of a cytosolic Hsp70 gene from Ulva pertusa (Ulvophyceae, Chlorophyta)[J]. JOURNAL OF APPLIED PHYCOLOGY,2011,23(4):681-690.
APA	Fu, Wandong,Shuai, Li,Yao, Jianting,Zheng, Bin,Zhong, Mingjie,&Duan, Delin.(2011).Molecular cloning and expression analysis of a cytosolic Hsp70 gene from Ulva pertusa (Ulvophyceae, Chlorophyta).JOURNAL OF APPLIED PHYCOLOGY,23(4),681-690.
MLA	Fu, Wandong,et al."Molecular cloning and expression analysis of a cytosolic Hsp70 gene from Ulva pertusa (Ulvophyceae, Chlorophyta)".JOURNAL OF APPLIED PHYCOLOGY 23.4(2011):681-690.

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