合浦珠母贝骨形态发生蛋白10(BMP10)基因的克隆与表达分析

Molecular cloning and expression analysis of BMP10 gene from pearl oyster( Pinctada fucata)

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 functions of inducing bone formation as well as regulating embryo development, histogeny and reproduction. However, there are only a few researches on BMPs in molluscs. The BMPIO gene was cloned from pearl oyster （Pinctadafucata） and thus named as PfBMPIO. The full length cDNA of PfBMPIO is 1 752 bp, including an open reading frame （ORF） of 1 344 bp which encodes 447 amino acids. The protein sequence includes an N-terminal signal peptide, a pro-domain and a mature protein region. The mature protein contains transforming growth factor-β （TGF-β） domain which has seven cysteine resi- dues. PfBMPIO expressed higher at the trochophore stage and metamorphosis during the larval development as indicated by qPCR, suggesting that it may involve in larval developmental regulation of trochophore stage and metamorphosis. Compared with other tissues, PfBMPIO showed high relative expression level in the gill and mantle, and rose in 24 h after shell notching, indicating that it probably involves in shell formation. The study provides references for functional study of BMP family genes and mechanism study of shell formation.