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Identification of calconectin, a calcium-binding protein specifically expressed by the mantle of Pinctada
D Duplat1, M Puisségur, L Bédouet
1USM 0401, UMR CNRS 5178 Biologie des Organismes Marins et Ecosystèmes, Muséum National d'Histoire Naturelle, 7, rue Cuvier, 75005 Paris, France. duplat@mnhn.fr
Abstract:
Nacre or mother-of-pearl in the shell of Pinctada margaritifera is composed of 95-99% calcium carbonate and 1-5% organic matrix. In this study, we developed an original technique to characterize the genes differentially expressed in nacre-forming cells (NFC) by combining suppression subtractive hybridization (SSH), to establish a cDNA subtractive library, with rapid amplification of cDNA ends (RACE)-PCR. Seventy-two specific cDNA sequences have been obtained so far. These include a protein containing two EF-hand Ca2+-binding domains which was completely sequenced after amplification by RACE-PCR. Its specific expression as well as the specificity of the SSH method was confirmed by semi-quantitative RT-PCR on NFC and mantle cells.
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