【摘要】 采用RT-PCR技术从美洲大蠊中扩增出变应原Cr-pII基因,并将其克隆至pMD18-T载体中。经序列测定和分析证实,所克隆的片段含有长度为1185bp的完整开放阅读框(ORF),与台湾株来源的Cr-pII(Per a1·0103)有95·9%的同源性。用NdeⅠ和NotⅠ将目的片段从T载体中切下,定向亚克隆入表达载体pET24a(+),然后成功转化大肠杆菌菌株BL21Star。经IPTG诱导得到45kDa的重组蛋白,Western-blot分析表明其具有良好的IgE结合活性。并通过生物信息学方法获得了目的蛋白的抗原表位和3D结构模型。更多还原

【Abstract】 The Cr-pII gene was amplified from Periplaneta Americana by RT-PCR and subsequently was cloned into the pMD18-T vector.Sequence analysis showed that this fragment contained a complete ORF of 1 185 bp and had 95.9% homology with Per a 1.010 3 that was cloned from Taiwan strain.The fragment was subcloned into expression plasmid pET24a(+),then the recombinant plasmid was transformed into E.coli BL21 Star for expression.After induced by IPTG,45kDa recombinant protein was obtained.Western-blot indicated it has good IgE binding potency.The antigen epitope and 3D structure model of Cr-pII were predicted by biological information methods.更多还原