Cloning and characterization of metallothionein gene in ayu Plecoglossus altivelis.
Metallothionein (MT) has been used widely as a potential molecular marker to detect the deleterious effects of heavy metals in aquatic ecosystem. Here we exposed ayu, Plecoglossus altivelis, to zinc (Zn) and tested the distribution as well as the induction of MT in various tissues such as liver, kidney, intestine and stomach. MT induction was significant in liver tissue, followed by kidney and intestine, whereas no induction was detected in stomach. The gene encoding ayu MT was successfully cloned and characterized. Complete nucleotide sequencing and analysis of the 4.5 kb DNA fragment containing the ayu MT gene revealed that the gene has three exons interrupted by two introns, a 5'-flanking region of about 2.5 kb and about 1.6 kb of 3'-flanking region. In grouper heart and kidney cells, the 2.5 kb promoter containing eight metal responsive elements (MREs), two hepatic nuclear factor 5 responsive elements (HNF5REs) and one cAMP responsive element (CRE) had the highest reporter activity.
Lin
Chih-Hung
CH
Molecular Genetics Laboratory, 336, Institute of Zoology (44), Academia Sinica, 128, Academia Road Section 2, NanKang, Taipei 11529, Taiwan, ROC.
John
Joseph Abraham Christopher
JA
Ou
Li Wei
LW
Chen
Jun-Chang
JC
Lin
Cheng-Hui
CH
Chang
Chi-Yao
CY
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015415:Biomarkers; D015152:Blotting, Northern; D017931:DNA Primers; D040461:Gene Components; D005786:Gene Expression Regulation / Q000235:genetics*; D008668:Metallothionein / Q000235:genetics*; D008969:Molecular Sequence Data; D036441:Osmeriformes / Q000235:genetics*; D010802:Phylogeny; D011401:Promoter Regions, Genetic / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D015415:Biomarkers; D017931:DNA Primers; D008668:Metallothionein