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骨形态发生蛋白(bone morphogenetic proteins,BMPs)具有成骨活性,在胚胎发育、组织发生、生殖调控等方面也有重要作用,但在贝类中研究还很少。研究克隆了合浦珠母贝(Pinctada fucata)BMP10基因,命名为Pf BMP10。该基因c DNA长1 752 bp,包含开放阅读框(ORF)1 344 bp,编码447个氨基酸。编码蛋白包含一个N端信号肽、一个前结构域和成熟蛋白区。成熟区包含转化生长因子β(transforming growth factor beta,TGF-β)结构域,其中含7个半胱氨酸残基。不同发育时期表达分析显示Pf BMP10高表达于担轮期和变态期,提示其可能参与幼体担轮期、变态期的发育调控。组织表达分析显示其在鳃和外套膜高表达,贝壳缺刻实验显示缺刻处理24h后其在鳃和外套膜中的表达量均显著上升,表明Pf BMP10可能与贝壳形成相关。研究结果可为BMP家族基因功能及贝壳形成机理研究提供参考。
Bone morphogenetic proteins (BMPs) have osteogenic activity and play important roles in embryonic development, tissue formation and reproductive regulation. However, few studies have been done in shellfish. The cloned Pinctada fucata BMP10 gene was named Pf BMP10. The cDNA of this gene is 1 752 bp in length and contains 1 344 bp of open reading frame (ORF) encoding 447 amino acids. The encoded protein contains an N-terminal signal peptide, a pre-domain and a mature protein region. The mature region contains a transforming growth factor β (TGF-β) domain containing 7 cysteine residues. Expression analysis at different developmental stages showed that Pf BMP10 overexpression was in the stage of bearing and metamorphosis, suggesting that Pf BMP10 may be involved in the developmental regulation of larvae during the roping and metamorphosis. Tissue expression analysis showed that it was highly expressed in the gill and mantle, and shell scoring experiments showed that the expression of GFP and mantle increased significantly after 24 hours of nicking treatment, indicating that Pf BMP10 may be related to the formation of the shell. The results provide a reference for the study of BMP gene function and shell formation mechanism.