采用同源克隆和锚定PCR技术,从异色瓢虫Harmoniaaxyridis(Pallas)中克隆到热休克蛋白HSP90基因的eDNA全序列(Genbanknumber:FJ501962)。eDNA全长2480bp,包含3’非编码区域(UTR)为200bp和5’UTR为126bp,开放阅读框(ORF)长2154bp,编码717个氨基酸。预测的相对分子质量为82230,理论等电点为4.96,无糖基化位点、跨膜结构和信号肽。在N端具有HSP90基因保守的ATPase结构,含有HSP90家族的C末端的保守序列EEVD。与赤拟谷盗Triboliumcastaneurn相比较,同源性高达90%,系统发育分析也表明两者的亲缘关系最近。HaaHSP90基因的克隆和比较分析为进一步深入研究异色瓢虫的抗逆机理及其进化具有重要意义。
The heat shock protein 90 (HSP90) gene from Harmon& axyridis (Pallas) was cloned using techniques of homological cloning and anchored PCR. The full length of cDNA sequence is 2 480 bp (Genbank number: FJ501962), containing a 3'UTR (untranslated region) of 200 bp, a 5'UTR of 126 bp, and an ORF of 2 154 bp which encodes a polypeptide of 717 amino acids with an estimated molecular weight of 82 230 and pl of 4.96. There are no glycosylation sites, putative transmembrane domain and signature peptide. It contains ATPase conservative domain of (HSP90) in N-site and EEVD signature sequence on C-site. Compared with Tribolium castaneum, they are 90% identities and Phylogenetic analysis shows that these two species are most closely related. Cloning and comparison analysis of HSP90 genes from Harmonia axyridis (Pallas) could be useful in the studies of anti-stress mechanism, tolerance improvement breeding and evolution for these animals.