Molecular Cloning and Expression Analysis of a Cytosolic Hsp70 gene from Laminaria japonica (Laminariaceae, Phaeophyta)

doi:10.1007/s10126-009-9188-z

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	Molecular Cloning and Expression Analysis of a Cytosolic Hsp70 gene from Laminaria japonica (Laminariaceae, Phaeophyta)
	Fu, Wandong 1,2; Yao, Jianting 1; Wang, Xiuliang 1; Liu, Fuli 1,2; Fu, Gang 1; Duan, Delin 1
	2009-12-01
发表期刊	MARINE BIOTECHNOLOGY
ISSN	1436-2228
卷号	11 期号:6 页码:738-747
文章类型	Article
摘要	In this study, a full-length cytosolic heat shock protein 70 complementary DNA (cDNA) of Laminaria japonica (designated as LJHsp70) was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR) coupled with rapid amplification of cDNA ends. The full length of LJHsp70 cDNA was 2,918 bp, with a 5' untranslated region of 248 bp, a 3' untranslated region of 696 bp, and an open reading frame of 1,974 bp encoding a polypeptide of 657 amino acids with an estimated molecular mass of 72.03 kDa and an estimated isoelectric point of 4.97. There was highly repeated sequence of CAA in 5' untranslated region of LJHsp70. The result of phylogenetic tree of Hsp70s, the BLAST program, analysis and cytosolic Hsp70-specific motif of LJHsp70 verified that the cloned LJHsp70 belonged to cytosolic Hsp70 family. Three typical Hsp70 signature motifs were detected in LJHsp70 by InterPro analysis. Under different stress conditions, messenger RNA (mRNA) expression levels of LJHsp70 were quantified by quantitative RT-PCR. To L. japonica sporophytes kept in different temperatures for 1 h, the expression level of LJHsp70 at 30A degrees C was highest and twofold higher than that at 10A degrees C. To L. japonica sporophytes kept at 25A degrees C for different times, the mRNA expression level of LJHsp70 reached a maximum level after 7 h and then dropped progressively. The expression level of LJHsp70 at 0 or 5aEuro degrees salt concentration for 2 h was twofold higher than that at 30aEuro degrees salt concentration for 2 h. The results showed that LJHsp70 may be a kind of potential biomarker used to monitor environment conditions.; In this study, a full-length cytosolic heat shock protein 70 complementary DNA (cDNA) of Laminaria japonica (designated as LJHsp70) was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR) coupled with rapid amplification of cDNA ends. The full length of LJHsp70 cDNA was 2,918 bp, with a 5' untranslated region of 248 bp, a 3' untranslated region of 696 bp, and an open reading frame of 1,974 bp encoding a polypeptide of 657 amino acids with an estimated molecular mass of 72.03 kDa and an estimated isoelectric point of 4.97. There was highly repeated sequence of CAA in 5' untranslated region of LJHsp70. The result of phylogenetic tree of Hsp70s, the BLAST program, analysis and cytosolic Hsp70-specific motif of LJHsp70 verified that the cloned LJHsp70 belonged to cytosolic Hsp70 family. Three typical Hsp70 signature motifs were detected in LJHsp70 by InterPro analysis. Under different stress conditions, messenger RNA (mRNA) expression levels of LJHsp70 were quantified by quantitative RT-PCR. To L. japonica sporophytes kept in different temperatures for 1 h, the expression level of LJHsp70 at 30A degrees C was highest and twofold higher than that at 10A degrees C. To L. japonica sporophytes kept at 25A degrees C for different times, the mRNA expression level of LJHsp70 reached a maximum level after 7 h and then dropped progressively. The expression level of LJHsp70 at 0 or 5aEuro degrees salt concentration for 2 h was twofold higher than that at 30aEuro degrees salt concentration for 2 h. The results showed that LJHsp70 may be a kind of potential biomarker used to monitor environment conditions.
关键词	Laminaria Japonica Heat Shock Protein 70 Cdna Cloning Qrt-pcr Mrna Expression
学科领域	Biotechnology & Applied Microbiology ; Marine & Freshwater Biology
DOI	10.1007/s10126-009-9188-z
URL	查看原文
收录类别	SCI
语种	英语
WOS记录号	WOS:000271644100008
引用统计	被引频次：31[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/337002/2902
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.Chinese Acad Sci, Grad Sch, Beijing 100039, 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,Yao, Jianting,Wang, Xiuliang,et al. Molecular Cloning and Expression Analysis of a Cytosolic Hsp70 gene from Laminaria japonica (Laminariaceae, Phaeophyta)[J]. MARINE BIOTECHNOLOGY,2009,11(6):738-747.
APA	Fu, Wandong,Yao, Jianting,Wang, Xiuliang,Liu, Fuli,Fu, Gang,&Duan, Delin.(2009).Molecular Cloning and Expression Analysis of a Cytosolic Hsp70 gene from Laminaria japonica (Laminariaceae, Phaeophyta).MARINE BIOTECHNOLOGY,11(6),738-747.
MLA	Fu, Wandong,et al."Molecular Cloning and Expression Analysis of a Cytosolic Hsp70 gene from Laminaria japonica (Laminariaceae, Phaeophyta)".MARINE BIOTECHNOLOGY 11.6(2009):738-747.

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