柞蚕热休克蛋白70基因的克隆和序列分析

摘要

HSP70 protein is stimulated by heat and other factors which induced the protein and it is a member of heat shock protein family, also the most important one.Using RT-PCR, we cloned Tussah (Antheraea pernyi) heat shock protein 70 gene (HSP70) of the ORF sequence.Bioinformatic analysis showed that the sequence encodes 650 amino acids with structural features of HSP70 conservative, and Cecropia moth (Antheraea yamamai), silkworm (Bombyx mor ), cabbage armyworm ( Mamestra brassica), cotton bollworm ( Heliothis viriplaca), beet armyworm ( Spodoptera exigua), tobacco armyworm (Manduca sexta ), Hymenoptera, parasitic wasps (Cotesia rubecula ) homology were 95.7 ％, 78.5 ％, 76.1％, 77.3 ％, 76.6 ％, 74.7 ％, 65.9 ％.According to their structure, we constructed a phylogenetic tree, and further established the kinship between them.%HSPTO蛋白是受热等因素刺激后而诱导产生的蛋白质,是热休克蛋白家族中最重要的一员.采用RTPCR方法克隆了柞蚕(Antherosa pernyi)热休克蛋白70基因(HSP70)的ORF序列(GenBank登录号:GU945199),该片段的序列长度为1905 bp.生物信息学分析表明,该序列共编码634个氨基酸,预测蛋白的等电点和分子量大小分别为5.62 kD和69.5 kD.具有HSPTO的保守性结构特征,与天蚕(Antheraea yamamai)、家蚕(Bombyx mor)、甘蓝夜蛾(Mamestra brassicae)、棉铃虫(Heliothis viriplaca)、甜菜夜蛾(Spodoptera exigua)、烟草夜蛾(Mandusa sextal)、膜翅目寄生蜂(Cotesia rubecula)的同源性分别为95.7%、78.5%、76.1%、77.3%、76.6%、74.7%、65.9%.根据它们的一级结构构建了系统进化树,进一步确立了它们之间的亲缘关系.