中华绒螯蟹14-3-3ζ基因cDNA全长克隆与序列分析

cDNA Cloning,Characterization and Sequence Analyses of 14-3-3ζGene in Eriocheir sinensis

14-3-3蛋白可以与不同蛋白质结合从而参与多种磷酸化调节的信号通路,并在生物代谢、信号转导、细胞周期控制、凋亡、蛋白质运输、转录、应激反应等方面发挥重要作用.本文用SMART RACE技术克隆获得了中华绒螯蟹14-3-3ζ基因的全长cDNA序列,14-3-3ζ基因cDNA全长为1081 bp,包含一个744 bp的开放阅读框,其编码一个28.0 kDa的含247个氨基酸的多肽,编码的氨基酸序列包含保守的14-3-3结构域.多序列比对、系统发生关系重建和其他生物信息学分析表明中华绒螯蟹14-3-3ζ为非分泌型蛋白,不具有跨膜区域,定位于细胞质中,其分子量与已知的其他物种的14-3-3ζ蛋白分子量相近.中华绒螯蟹的14-3-3ζ序列与南美白对虾(Penaeus vannamei)、拟穴青蟹(Scylla paramamosain)等甲壳类动物的14-3-3ζ基因高度同源.选择压力分析表明中华绒螯蟹14-3-3ζ受到纯化选择,其功能可能高度保守.

14-3-3 proteins can bind to different proteins to participate in a variety of phosphorylation regulatory signaling pathways,and play an important role in biological metabolism,signal transduction,cell cycle control,apoptosis,protein transport,transcription,stress response and other aspects.The full-length cDNA of 14-3-3ζgene from the Chinese mitten crab Eriocheir sinensis was cloned and identified using SMART RACE technique.It is consisted of 1081 nucleotides with an open-reading frame of 744 bp,which encoding 247 amino acids(28.0 kDa)and containing 14-3-3 domain.Sequence alignment,phylogenetic tree construction and some bioinformatics analyses revealed that 14-3-3ζis non-secretory and no-transmembrane protein,which is probable located in cytoplasm.The molecular weight of 14-3-3ζis similar than that of others 14-3-3ζisoforms.The sequence of 14-3-3ζin E.sinensis is highly homologous to that in the reported crustaceans such as Penaeus vannamei and Scylla paramamosain.Moreover,we observed a strong signature of purifying selection between the comparison of orthologous 14-3-3ζgene pairs of E.sinensis.The results suggest a conservative function across crustaceans.