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Molecular cloning and characterization of the 5'-flanking regulatory region of the carbonic anhydrase nacrein gene of the pearl oyster Pinctada fucata and its expression.

ABSTRACT: The carbonic anhydrase nacrein participates in the formation of the nacreous or prismatic layer of Pinctada fucata. We isolated a genomic clone containing the nacrein gene and cloned the 5'-flanking region. Within the 1336 bp 5' flanking region, we identified putative cis-acting elements, including the TATA box (TATAAAA) at -82 bp, and AP1 (-819 bp) and Oct-1 (-1244 bp) binding sites. In addition to the mantle, the nacrein gene is also expressed in the adductor muscle, liver, and foot. These results showed that nacrein not only takes part in the formation of the hard tissue but also might be involved in acid-base balance, ion transport, and maintenance of ionic concentration. In vitro transcription experiments showed that the addition of human c-jun activates transcription from the nacrein promoter. This is the first report of a promoter from a gene that controls the formation of the hard tissue of mollusk shells.

SUBMITTER: Miyashita T

PROVIDER: S-EPMC3439604 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature

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Molecular cloning and characterization of the 5'-flanking regulatory region of the carbonic anhydrase nacrein gene of the pearl oyster Pinctada fucata and its expression.

Miyashita Tomoyuki T Takami Akiko A Takagi Ryosuke R

Biochemical genetics 20120510 9-10

The carbonic anhydrase nacrein participates in the formation of the nacreous or prismatic layer of Pinctada fucata. We isolated a genomic clone containing the nacrein gene and cloned the 5'-flanking region. Within the 1336 bp 5' flanking region, we identified putative cis-acting elements, including the TATA box (TATAAAA) at -82 bp, and AP1 (-819 bp) and Oct-1 (-1244 bp) binding sites. In addition to the mantle, the nacrein gene is also expressed in the adductor muscle, liver, and foot. These res ...[more]

PMID: 22573138

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Project description:BackgroundThe most critical step in the pearl formation during aquaculture is issued to the proliferation and differentiation of outer epithelial cells of mantle graft into pearl sac. This pearl sac secretes various matrix proteins to produce pearls by a complex physiological process which has not been well-understood yet. Here, we aimed to unravel the genes involved in the development of pearl sac and pearl, and the sequential expression patterns of different shell matrix proteins secreted from the pearl sac during pearl formation by pearl oyster Pinctada fucata using high-throughput transcriptome profiling.ResultsPrincipal component analysis (PCA) showed clearly different gene expression profiles between earlier (before 1 week) and later stages (1 week to 3 months) of grafting. Immune-related genes were highly expressed between 0 h - 24 h (donor dependent) and 48 h - 1 w (host dependent), and in the course of wound healing process pearl sac was developed by two weeks of graft transplantation. Moreover, for the first time, we identified some stem cell marker genes including ABCG2, SOX2, MEF2A, HES1, MET, NRP1, ESR1, STAT6, PAX2, FZD1 and PROM1 that were expressed differentially during the formation of pearl sac. The expression profiling of 192 biomineralization-related genes demonstrated that most of the shell matrix proteins (SMPs) involved in prismatic layer formation were first up-regulated and then gradually down-regulated indicating their involvement in the development of pearl sac and the onset of pearl mineralization. Most of the nacreous layer forming SMPs were up-regulated at 2 weeks after the maturation of pearl sac. Nacrein, MSI7 and shematrin involved in both layer formation were highly expressed during 0 h - 24 h, down-regulated up to 1 week and then up-regulated again after accomplishment of pearl sac formation.ConclusionsUsing an RNA-seq approach we unraveled the expression pattern of the key genes involved in the development of pearl sac and pearl as a result of host immune response after grafting. These findings provide valuable information in understanding the molecular mechanism of pearl formation and immune response in P. fucata.

Dataset Information

Molecular cloning and characterization of the 5'-flanking regulatory region of the carbonic anhydrase nacrein gene of the pearl oyster Pinctada fucata and its expression.

Publications

Molecular cloning and characterization of the 5'-flanking regulatory region of the carbonic anhydrase nacrein gene of the pearl oyster Pinctada fucata and its expression.

Similar Datasets

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