Metallothionein-2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization

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	Metallothionein-2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization
	Gao, Dian 1,2,3; Wang, Gui Tang 1,2; Chen, Xing Tao 1,2,3; Nie, Pin 1,2
刊名	COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY
	2009
卷号	149 期号:1 页码:18-25
关键词	Expression Gene Mandarin fish Metallothionein
ISSN号	1532-0456
通讯作者	Wang, GT, Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hunan, Peoples R China
中文摘要	The metallothionein-2 (MT-2) gene was isolated from the mandarin fish, one of the most important industrial aquatic animals in China, by using rapid amplification of cDNA ends (RACE). The deduced amino acid sequence of MT-2 comprised 60 amino acids and showed approximately 62.3% identity to human metallothionein. Its promoter region was amplified by thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR). The MT-2 gene consists of 3 exons and 2 introns, extending approximately 900 bp of genomic sequence. Phylogenetic analysis clearly demonstrated that MT-2 formed a clade with fish metallothionein. The promoter region contained 5 putative metal-regulatory elements (MREs) and 1 TATA box. Real-time quantitative RT-PCR analysis revealed that MT-2 transcripts were significantly increased in the brain and gills and were stable in the muscles, liver, and trunk kidney in Cd2+-stimulated fish. Western blotting analysis demonstrated that the protein of the MT-2 gene was expressed mainly in the gills, liver, heart, trunk kidney, muscle, and intestine; it was weakly detected in the brain and head kidney. Moreover, the MT-2 protein was immunohistochemically detected in the cytoplasm in the liver and trunk kidney. All the above results revealed that the mandarin fish MT-2 would be a useful biomarker for metal pollution. (C) 2008 Published by Elsevier Inc.
英文摘要	The metallothionein-2 (MT-2) gene was isolated from the mandarin fish, one of the most important industrial aquatic animals in China, by using rapid amplification of cDNA ends (RACE). The deduced amino acid sequence of MT-2 comprised 60 amino acids and showed approximately 62.3% identity to human metallothionein. Its promoter region was amplified by thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR). The MT-2 gene consists of 3 exons and 2 introns, extending approximately 900 bp of genomic sequence. Phylogenetic analysis clearly demonstrated that MT-2 formed a clade with fish metallothionein. The promoter region contained 5 putative metal-regulatory elements (MREs) and 1 TATA box. Real-time quantitative RT-PCR analysis revealed that MT-2 transcripts were significantly increased in the brain and gills and were stable in the muscles, liver, and trunk kidney in Cd(2+)-stimulated fish. Western blotting analysis demonstrated that the protein of the MT-2 gene was expressed mainly in the gills, liver, heart, trunk kidney, muscle, and intestine; it was weakly detected in the brain and head kidney. Moreover, the MT-2 protein was immunohistochemically detected in the cytoplasm in the liver and trunk kidney. All the above results revealed that the mandarin fish MT-2 would be a useful biomarker for metal pollution. (C) 2008 Published by Elsevier Inc.
学科主题	Biochemistry & Molecular Biology; Endocrinology & Metabolism; Toxicology; Zoology
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biochemistry & Molecular Biology ; Endocrinology & Metabolism ; Toxicology ; Zoology
研究领域[WOS]	Biochemistry & Molecular Biology ; Endocrinology & Metabolism ; Toxicology ; Zoology
关键词[WOS]	ASYMMETRIC INTERLACED PCR ; CARP CARASSIUS-CUVIERI ; TROUT SALMO-TRUTTA ; MESSENGER-RNA ; HEPATIC METALLOTHIONEIN ; PROMOTER REGION ; REGULATORY ELEMENTS ; PLATICHTHYS-FLESUS ; CADMIUM EXPOSURE ; METAL POLLUTION
收录类别	SCI
资助信息	Hubei Gongguan Project of Science and Technology [2007AA203A01]; National Basic Research Program [2002CB412308]
语种	英语
WOS记录号	WOS:000261936200002
公开日期	2010-10-13
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/152342/7934]
专题	水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文
作者单位	1.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hunan, Peoples R China 2.Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hunan, Peoples R China 3.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
推荐引用方式 GB/T 7714	Gao, Dian,Wang, Gui Tang,Chen, Xing Tao,et al. Metallothionein-2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization[J]. COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY,2009,149(1):18-25.
APA	Gao, Dian,Wang, Gui Tang,Chen, Xing Tao,&Nie, Pin.(2009).Metallothionein-2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization.COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY,149(1),18-25.
MLA	Gao, Dian,et al."Metallothionein-2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization".COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY 149.1(2009):18-25.