红笛鲷MAPKK5基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of MAPKK5 Gene in Humphead Snapper Lutjanus sanguineus

利用本实验室构建红笛鲷头肾cDNA文库中的MAPKK5基因EST序列,通过RACE PCR技术克隆红笛鲷MAPKK5基因cDNA全长(Ls-MAPKK5,登录号为KM657202)。序列分析结果显示,Ls-MAPKK5cDNA全长2632bp,其中开放阅读框1317bp,可编码438个氨基酸,预测其编码蛋白的分子量为49.29ku,理论等电点为6.12。SignalP 4.0、TMHMM Server 2.0和SoftBerry-Psite预测结果显示,Ls-MAPKK5没有信号肽和跨膜结构,含有22个磷酸化位点。氨基酸序列分析显示LsMAPKK5具有保守的PB1与S_TKc结构域。利用MEGA5.0软件,根据邻位相连法构建MAPKK5系统进化树,结果显示Ls-MAPKK5与雀鲷、吉富罗非鱼该蛋白有较近的亲缘关系。

In present study,MAPKK5（Ls-MAPKK5,GenBank accession number KM657202）was cloned in humphead snapper（Lutjanus sanguineus）using RACE method based on the corresponding EST,which was obtained from cDNA library constructed using head kidney of humphead snapper in our laboratory.The full-length cDNA of Ls-MAPKK5 was 2632bp containing an open reading frame of 1317 bp encoding438amino acids with an estimated molecular weight of 49.29 ku and a theoretical pI of 6.12.The prognosis by SignalP 4.0,TMHMM Server 2.0and SoftBerry-Psite revealed that the deduced amino acid of Ls-MAPKK5 did not contain a signal peptide or a transmembranous region,but had 22 phosphorylation sites.Amino acid alignment showed that Ls-MAPKK5 possessed a PB1 and S_TKc domain.Phylogenetic tree by NJ method using MEGA5.0indicated that Ls-MAPKK5 protein clustered closely with Stegastes partitus and Nile tilapia Oreochromis niloticus MAPKK5 protein.