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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
Objective To amplify the DNA fragment of the heat shock protein A 35 1bp of H. pylori and clone it into the plasmid pET - 2 8a (+) for efficient expression. Methods The amplified fragment was sequenced and analyzed by GoldKey software. The pET - 2 8a (+) system was used to express heat shock proteins in the recipient strain BL2 1 (DE3) pLysS. Results The sequence of the expressed protein was similar to the reported heat shock protein A sequence. After induced by IPTG, SDS - PAGE and TLC analysis showed that the relative molecular mass of the expressed foreign protein was 180 000, which was in the form of inclusion body. The expressed product accounted for 64% of the total bacterial protein. Conclusion HspA is most likely to be a candidate antigenic component in H. pv vaccine