【摘要】 目的　扩增幽门杆菌热休克蛋白A 35 1bp的DNA片段 ,并将其克隆到pET - 2 8a(+ )质粒中高效表达。方法　用PCR方法扩增的片段经测序后 ,用GoldKey分析软件进行序列分析 ,应用pET - 2 8a(+ )系统在受体菌BL2 1(DE3)pLysS中表达热休克蛋白。结果　该序列表达的蛋白与已报道的热休克蛋白A序列相似。重组菌株用IPTG诱导后 ,经SDS PAGE和薄层扫描分析 ,表达的外源蛋白的相对分子质量为 180 0 0 ,以包涵体的形式存在 ,表达产物占菌体总蛋白的 6 4%。结论　HspA是最有可能做为H .p疫苗中的候选抗原成分更多还原

【Abstract】 Objective To amplify a 351bp of DNA fragment of heat shock protein A from Helicobacter pylori, then clone it into plasmid vector pET28-a (+) for expression. Methods The amino acid composition of the fragment amplified by PCR was deduced by Gold Key software after sequencing. The recombinant plasmid was transformed to E. coli BL21(DE3)pLysS for expressing heat shock protein A under the induction of IPTG.Results The sequences of DNA and deduced amino acid of the expressed product was similar to those of heat shock protein A reported. SDS PAGE showed that the expressed heat shock protein A with a molecular weight of 18000 was inclusion body, and thin layer scanning proved that about 64% of total somatic protein was expressed. Conclusion HspA might be a candidate antigen for preparing H.p vaccine.更多还原