title: The mining of pearl formation genes in pearl oyster Pinctada fucata by cDNA suppression subtractive hybridization.
abstract: Recent researches revealed the regional preference of biomineralization gene transcription in the pearl oyster Pinctada fucata: it transcribed mainly the genes responsible for nacre secretion in mantle pallial, whereas the ones regulating calcite shells expressed in mantle edge. This study took use of this character and constructed the forward and reverse suppression subtractive hybridization (SSH) cDNA libraries. A total of 669 cDNA clones were sequenced and 360 expressed sequence tags (ESTs) greater than 100 bp were generated. Functional annotation associated 95 ESTs with specific functions, and 79 among them were identified from P. fucata at the first time. In the forward SSH cDNA library, it recognized mass amount of nacre protein genes, biomineralization genes dominantly expressed in the mantle pallial, calcium-ion-binding genes, and other biomineralization-related genes important for pearl formation. Real-time PCR showed that all the examined genes were distributed in oyster mantle tissues with a consistence to the SSH design. The detection of their RNA transcripts in pearl sac confirmed that the identified genes were certainly involved in pearl formation. Therefore, the data from this work will initiate a new round of pearl formation gene study and shed new insights into molluscan biomineralization.
authors.affiliation: Department of Aquatic Bioscience, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan.
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.forename: Shigeharu
authors.initials: S
authors.lastname: Kinoshita
authors.forename: Naoko
authors.initials: N
authors.lastname: Nomura
authors.forename: Chihiro
authors.initials: C
authors.lastname: Riho
authors.forename: Kaoru
authors.initials: K
authors.lastname: Maeyama
authors.forename: Kiyohito
authors.initials: K
authors.lastname: Nagai
authors.forename: Shugo
authors.initials: S
authors.lastname: Watabe
mesh_terms: D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D020224:Expressed Sequence Tags; D015723:Gene Library; D007564:Japan; D008969:Molecular Sequence Data; D060734:Nacre / Q000235:genetics*; D009693:Nucleic Acid Hybridization / Q000379:methods; D049911:Pinctada / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D002135:Calcium-Binding Proteins; D017931:DNA Primers; D060734:Nacre