东北林蛙PDK1基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of PDK1 Gene in Rana dybowskii

PDK1是PI3K/Akt信号通路中的一个重要的激酶。在哺乳动物中,PI3K/Akt信号通路是一条保守的通路,通过激活下游的因子对糖代谢、脂肪代谢以及蛋白质代谢,细胞的增殖、分化、凋亡等起调节控制作用。本研究基于东北林蛙转录组数据,从中鉴定出PDK1基因3条Unigene序列,并通过RACE法克隆了PDK1基因的全长cDNA,命名为rdPDK1,其全长为1 990 bp,开放阅读框长1 674 bp,编码557个氨基酸。进行生物信息学分析,推测该基因编码的蛋白质相对分子质量63.7 kD,等电点为7.21。该蛋白第82~344个和443~549个氨基酸是2段保守结构域STKc激酶域和PH调节结构域。同源性分析表明rdPDK1蛋白与HsPDK1蛋白氨基酸序列一致率为79%,相似率为100%;与MmPDK1蛋白氨基酸序列一致率为80%,相似率为100%。本研究结果有望为进一步研究东北林蛙PI3K/Akt信号通路中PDK1基因的特性及功能奠定基础。

PDK1 is an important kinase in the PI3K/Akt signal pathway. The PI3K/Akt signal pathway is a conserved pathway in mammals which,by activating downstream factors,regulates the metabolism of sugar,fat,and protein as well as cell differentiation,proliferation,and apoptosis.In this study,we identified three Unigene sequences of PDK1 gene according to the transcriptome database and cloned the full-length cDNA of PDK1 gene by RACE. The full-length cDNA of PDK1 was named rdPDK1. The full-length cDNA sequence was 1 990 bp and the open reading frame was 1 674 bp,which encoded a protein of 557 amino acids. Bioinformatics analysis showed that the protein encoded by this gene had a relative molecular mass of 63. 7 kD and an isoelectric point of 7. 21. The amino acids 82 to 344 and 443 to 549 of the protein are two conserved domains STKc and PH,respectively. The identity of RaPDK1 protein with HsPDK1 protein was 79%,and the similarity rate was 100%. The identity of RaPDK1 protein with MmPDK1 protein was 80% and the similarity rate was 100%. The results may provide a basis for the further study of the characterization and function of PDK1 gene in Rana dybowskii PI3K/Akt signal pathway.