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Cloning, characterization, expression, and copper sensitivity of the metallothionein-1 gene in the Chinese mitten crab, Eriocheir sinensis
Ren F.; Jiang H.; Sun J.; He L.; Li W.; Wang Y.; Wang Q.
2011-04-01
Source PublicationMolecular Biology Reports
ISSN03014851 15734978
Volume38Issue:4Pages:2383-2393
AbstractA full-length metallothionein-1(MT-1) cDNA was cloned from the Chinese mitten crab, Eriocheir sinensis, based upon the hepatopancreas cDNA library. The full-length cDNA contained a single 180 bp open reading frame that encoded a 59 amino acid protein. The deduced amino acid sequence was cysteine (Cys)-rich, with residues observed in patterns characteristic of other reported MTs: Cys-X-Cys, Cys-X-X-Cys, or Cys-X-X-X-Cys. Gene structure obtained via PCR yielded a 3816 bp gene, which was comprised of three exons and two introns arranged in a "3 + 2" pattern. The cloned 50flanking region (1,735 bp) contained several predicted binding sites, which included MREs, AP-1, SP1, USF, GATA, HNF-1, and HSF. MT-1 mRNA expression analysis revealed that while levels were highest in the hepatopancreas, expression was abundant in testis and thoracic ganglia, moderate in intestine (P<0.05), and weak in other tissues (P<0.05). MT-1 mRNA expression exhibited reproductive variation in the male, with levels approximately tenfold greater in August, during seasonal gonadal maturation, compared to other times of the year. Cu exposure via tank water (0-1 mg/l for 7 days) resulted in a dose-dependent bell curve response in MT-1 mRNA expression, with peak expression observed after exposure to 0.1 mg/l Cu. A time course experiment (0.1 mg/l Cu over 9 days) revealed MT-1 mRNA expression peaked sharply on day 5 before gradually decreasing with prolonged exposure. In the present report, we provide sequence analysis of the first MT-1 gene cloned in E. sinensis, and evidence that its physiological and toxicological regulation is evolutionary conserved. © Springer Science+Business Media B.V. 2010.
KeywordCloning Cu2+ induced expression Eriocheir sinensis Metallothionein-1 gene
DOI10.1007/s11033-010-0372-z
URLView the original
Language英語
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Cited Times [WOS]:14   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionUniversity of Macau
AffiliationEast China Normal University
Recommended Citation
GB/T 7714
Ren F.,Jiang H.,Sun J.,et al. Cloning, characterization, expression, and copper sensitivity of the metallothionein-1 gene in the Chinese mitten crab, Eriocheir sinensis[J]. Molecular Biology Reports,2011,38(4):2383-2393.
APA Ren F..,Jiang H..,Sun J..,He L..,Li W..,...&Wang Q..(2011).Cloning, characterization, expression, and copper sensitivity of the metallothionein-1 gene in the Chinese mitten crab, Eriocheir sinensis.Molecular Biology Reports,38(4),2383-2393.
MLA Ren F.,et al."Cloning, characterization, expression, and copper sensitivity of the metallothionein-1 gene in the Chinese mitten crab, Eriocheir sinensis".Molecular Biology Reports 38.4(2011):2383-2393.
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