Li Ming-Feng, Ling Zhen, Zhang Ying-Chun, Ma Ai-Ying, Sun Jian-Xin, Shi Hui-Juan, Wu Xiang-Fu
Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 1999;31(3):264-268.
Helicobacter pylori is the causative agent of gastritis and peptic ulcer in human. The heat-shock protein A (HspA)may stimulate the immunoresponse protecting human body against challenge of H.pylori. The gene encoding the structural A subunit of H.pylori heat-shock protein was amplified from H.pylori chromosomal DNA by PCR, and was inserted in the prokaryotic expression vector pET-22b(+), and then was transformed into the BL-21(DE3)E.coli strain to express the HspA recombinant protein. HspA gene was measured to be 354 base pairs, and the recombinant protein gene encoded polypeptides of 118 amino acid residues, corres-ponding to calculated molecular weight of 15 kD. Western blot analysis of HspA recombinant protein was confirmed that it could be specifically recognized by the serum of H.pylori-infected patients, and could also be re-cognized by the serum of immunized Balb/c mice with HspA itself. This result suggests that HspA may be an effective protein vaccine for prevention and treatment of the infection of H.pylori.
幽门螺杆菌是人类胃炎和消化性溃疡的病原体。热休克蛋白A(HspA)可能刺激免疫反应,保护人体免受幽门螺杆菌的攻击。通过PCR从幽门螺杆菌染色体DNA中扩增出编码幽门螺杆菌热休克蛋白结构A亚基的基因,并将其插入原核表达载体pET-22b(+)中,然后转化到BL-21(DE3)大肠杆菌菌株中表达HspA重组蛋白。测得HspA基因有354个碱基对,重组蛋白基因编码118个氨基酸残基的多肽,对应计算分子量为15kD。对HspA重组蛋白的Western印迹分析证实,它能被幽门螺杆菌感染患者的血清特异性识别,也能被用HspA自身免疫的Balb/c小鼠血清识别。该结果表明,HspA可能是预防和治疗幽门螺杆菌感染的有效蛋白疫苗。