【摘要】 从幽门螺杆菌染色体DNA ,用PCR方法扩增得到了热休克蛋白 70基因。序列分析表明 ,我国Hp临床分离株Y2 的热休克蛋白 70基因与经全基因组序列测定的两株幽门螺杆菌 2 6 6 95和J99有高度同源性。将该基因克隆到融合分泌表达载体 pMAL p2中 ,转化大肠杆菌 ,在IPTG诱导下表达出与预期大小相符的 113kD的融合表达蛋白。该蛋白质在 30℃诱导表达 5h后 ,可达到细菌周质总蛋白质的 19.4%。用免疫印迹分析表明 ,该融合表达蛋白能与抗幽门螺杆菌兔IgG发生特异性反应更多还原

【Abstract】 Heat-shock protein 70 gene (hsp70) was obtained by PCR method from Helico b acter pylori chromosomal DNA. Sequencing analysis exhibited that the hsp70 gene isolated from Hp Y 2 was highly homologous with the gene encoded in Heli cobacter pylori 26695 and J99, which had been sequenced for complete genome. T he hsp70 gene was recombined in vitro with fusion secretion expression v ector pMAL-p2 and was transformed into E.coli cells. The E.coli strains, containing hsp70 recombinant plasmid, expressed a 113 kD fusion protein which accounted f or 19.4% of the total bacterial periplasm protein after the induction with IPTG for 5 h at 3 0 ℃. The expressed fusion protein could react specifically with anti-Helicob acter pylori rabbit IgG, as proved by Western blot method.更多还原