Identification of calconectin, a calcium-binding protein specifically expressed by the mantle of Pinctada margaritifera |
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Authors: | Duplat D Puisségur M Bédouet L Rousseau M Boulzaguet H Milet C Sellos D Van Wormhoudt A Lopez E |
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Institution: | USM 0401, UMR CNRS 5178 Biologie des Organismes Marins et Ecosystèmes, Muséum National d'Histoire Naturelle, 7, rue Cuvier, 75005 Paris, France. duplat@mnhn.fr |
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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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Keywords: | RACE rapid amplification of cDNA ends NFC nacre-forming cells SSH suppression subtractive hybridization EDTA ethylenediaminetetraacetic acid CaRP calmodulin-related protein CaM calmodulin |
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