拟穴青蟹卵泡抑素相关蛋白基因的克隆和表达分析
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厦门大学 海洋与地球学院,厦门大学 海洋与地球学院,厦门大学 海洋与地球学院,厦门大学 海洋与地球学院,厦门大学 海洋与地球学院

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国家自然科学基金项目(40406030, 41076081)


Molecular cloning and expression analysis of the follistatin related protein gene in Scylla paramamosain
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college of marine and earth sciences, Xiamen University,college of marine and earth sciences, Xiamen University,college of marine and earth sciences, Xiamen University,college of marine and earth sciences, Xiamen University,college of marine and earth sciences, Xiamen University

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    摘要:

    卵泡抑素相关蛋白(follistatin related protein, FRP)是一种细胞外基质糖蛋白, 该蛋白参与细胞增殖、迁移、组织重塑、胚胎发育、细胞间相互作用等多种生理过程。迄今甲壳动物FRP的研究尚未见诸报道。实验采用RACE技术首次克隆了拟穴青蟹FRP基因部分cDNA序列。该序列长度1 948 bp, FRP肽段含有484个氨基酸残基, 包括KAZAL-FS结构域、EFh结构域和两个Ig结构域。系统进化树显示拟穴青蟹FRP基因的分子进化地位与其生物学分类地位一致。半定量PCR及荧光定量PCR结果表明, FRP基因在拟穴青蟹的胸神经团、脑和卵巢中表达。FRP基因在卵巢发育各期的表达量各不相同, 卵巢未发育期最高, 据此我们推测FRP具有抑制卵巢发育的作用。

    Abstract:

    Follistatin related protein (FRP) is an extracellular glycoprotein, involved in various physiological processes such as cell proliferation, migration, tissue remodeling, embryonic development, and cell-cell interaction. However, no FRP gene has been reported from crustaceans. In this study, we isolated FRP gene from mud crab, Scylla paramamosain by RACE (rapid amplification of cDNA ends). And we analyzed the expression of FRP transcripts in different tissues and ovarian development stages by semi-quantitative and quantitative RT-PCR. The obtained cDNA was 1 948 bp, with the 3’UTR of 493bp. The putative peptide of sp-FRP was composed of 484 amino acids, which was most similar to the red flour beetles Tribolium castaneum FRP (50%). Sp-FRP contained a KAZAL-FS domain, an EFh domain, and two immunoglobulin domains, suggesting that FRP may act as a scaffold protein linking the TGF-β family and other unknown proteins to modulate their functions. The phylogenetic analysis was consistent with traditional classification. Quantitative PCR showed that the expression of FRP was significantly higher in brain and thoracic ganglia than in ovary. In addition, the expression of FRP transcript was different during ovarian development, and much higher at undeveloped stage of ovary. FRP may play a suppression role in the ovarian development of the mud crab.

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解银洁,黄辉洋,叶海辉,巩 杰,金朱兴,李少菁.拟穴青蟹卵泡抑素相关蛋白基因的克隆和表达分析[J].水产学报,2012,36(8):1201~1208

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  • 收稿日期:2012-01-07
  • 最后修改日期:2012-05-04
  • 录用日期:2012-05-15
  • 在线发布日期: 2012-08-22
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