Identification of calconectin, a calcium-binding protein specifically expressed by the mantle of Pinctada margaritifera.
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.
Duplat
D
D
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
Puisségur
M
M
Bédouet
L
L
Rousseau
M
M
Boulzaguet
H
H
Milet
C
C
Sellos
D
D
Van Wormhoudt
A
A
Lopez
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000235:genetics; D018076:DNA, Complementary; D015870:Gene Expression; D008969:Molecular Sequence Data; D049911:Pinctada / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D002135:Calcium-Binding Proteins; D018076:DNA, Complementary