利用CODEHOP克隆意蜂Hsp90基因及生物信息分析
Cloning and Bioinformatics Analysis of Apis mellifera Ligustica Spin Hsp90 Gene

Hsp90(heat shock protein 90)是热休克蛋白家族中重要成员,在蜜蜂多种生理调控、应激及免疫反应中起到重要作用。本研究利用CODEHOP(consensus-degenerate hybrid oligonucleotide primers)法通过特定氨基酸序列设计简并引物首次克隆纯系意大利蜜蜂(Apis mellifera Ligustica Spin )Hsp90基因序列,对其进行生物信息学分析并构建系统进化树。结果表明蜜蜂Hsp90基因在进化中比较保守,经过比对意蜂与西方蜜蜂DNA序列同源性为99%;与大蜜蜂、小蜜蜂和熊蜂同源性分别为97%、97%和90%,该DNA片段编码一个由320个氨基酸组成的肽段,种属间氨基酸差异率为30%,氨基酸替代率在0.01到3.28之间。利用该方法设计简并引物,简并度低,特异性强,为蜜蜂未知功能基因克隆提供了便捷有效途径,开展Hsp90基因功能研究在遗传变异及种质资源保护中具有重要的意义。
Hsp90,a significant member of the heat shock protein family,plays an important role in a variety of physiological reactions related with stress and immune response.In this study,we cloned Hsp90 gene from Apis mellifera Ligustica Spin with a pair of consensus-degenerate hybrid oligonucleotide primers(CODEHOP).Bioinformatics analysis indicated that the Hsp90 gene is highly conservative.The homologies,compared with Hsp90 from Apis mellifera,Apis dorsata,Apis florae,Bombus terrestris are 99%,97%,96% and 90%,respectively.This fragment encodes a peptide of 320 amino acids.The amino acid variance ratio is 30% and the substitution rate is between 0.01 and 3.28.Functional study of Hsp90 is very important in genetic variation and germplasm conservation.Designing of degenerate primers provides a convenient and effective way to clone unknown genes of the bees.