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Metallothionein-2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization
Gao, Dian1,2,3; Wang, Gui Tang1,2; Chen, Xing Tao1,2,3; Nie, Pin1,2; Wang, GT, Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hunan, Peoples R China
2009
Source PublicationCOMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY
ISSN1532-0456
Volume149Issue:1Pages:18-25
AbstractThe 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.; 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.
SubtypeArticle
KeywordExpression Gene Mandarin Fish Metallothionein
Department[Gao, Dian; Wang, Gui Tang; Chen, Xing Tao; Nie, Pin] Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hunan, Peoples R China; [Gao, Dian; Wang, Gui Tang; Chen, Xing Tao; Nie, Pin] Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hunan, Peoples R China; [Gao, Dian; Chen, Xing Tao] Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
Subject AreaBiochemistry & Molecular Biology ; Endocrinology & Metabolism ; Toxicology ; Zoology
DOI10.1016/j.cbpc.2008.05.014
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Funding OrganizationHubei Gongguan Project of Science and Technology [2007AA203A01]; National Basic Research Program [2002CB412308] ; Hubei Gongguan Project of Science and Technology [2007AA203A01]; National Basic Research Program [2002CB412308] ; Hubei Gongguan Project of Science and Technology [2007AA203A01]; National Basic Research Program [2002CB412308] ; Hubei Gongguan Project of Science and Technology [2007AA203A01]; National Basic Research Program [2002CB412308]
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Endocrinology & Metabolism ; Toxicology ; Zoology
WOS SubjectBiochemistry & Molecular Biology ; Endocrinology & Metabolism ; Toxicology ; Zoology
WOS IDWOS:000261936200002
WOS KeywordASYMMETRIC INTERLACED PCR ; CARP CARASSIUS-CUVIERI ; TROUT SALMO-TRUTTA ; MESSENGER-RNA ; HEPATIC METALLOTHIONEIN ; PROMOTER REGION ; REGULATORY ELEMENTS ; PLATICHTHYS-FLESUS ; CADMIUM EXPOSURE ; METAL POLLUTION
Funding OrganizationHubei Gongguan Project of Science and Technology [2007AA203A01]; National Basic Research Program [2002CB412308] ; Hubei Gongguan Project of Science and Technology [2007AA203A01]; National Basic Research Program [2002CB412308] ; Hubei Gongguan Project of Science and Technology [2007AA203A01]; National Basic Research Program [2002CB412308] ; Hubei Gongguan Project of Science and Technology [2007AA203A01]; National Basic Research Program [2002CB412308]
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/7934
Collection期刊论文
Corresponding AuthorWang, GT, Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hunan, Peoples R China
Affiliation1.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
Recommended Citation
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,&Wang, GT, Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hunan, Peoples R China.(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.
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