4.6 Article

CDNA cloning and expression pattern of pi-class glutathione S-transferase in the freshwater bivalves Unio tumidus and Corbicula fluminea

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ELSEVIER SCIENCE INC
DOI: 10.1016/j.cca.2005.02.013

Keywords

Unio tumidus; Corbicula fluminea; pi-class GST; RT-PCR; degenerated primers; RACE-PCR; cDNA cloning; expression pattern

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Glutathione S-transferases (GSTs) are enzymes involved in major detoxification reactions of xenobiotics in many organisms. The aim of this work was the identification of GST transcripts in the freshwater bivalves Unio tumidus and Corbicula fluminea. We used degenerated primers designed in the highly conserved regions of GST to amplify the corresponding mRNA. Full-length coding sequences were obtained by 5' and 3' rapid amplification of cDNA ends. In the two species, the GST cDNAs identified encoded a protein of 205 amino acids. The comparison of the deduced amino acid sequences with GSTs from other species showed that the enzymes belong to the pi-class and the amino acids defining the binding sites of glutathione (G-site) and for xenobiotic substrates (H-site) are highly conserved. Specific amplifications of the GST mRNA from U tumidus and C fluminea were performed on the digestive gland, the excretory system and the gills. For each mussel, the results revealed that the pi-class GSTs are expressed at the same level in the three tissues. (c) 2005 Elsevier Inc. All rights reserved.

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