幽门螺杆菌热休克蛋白A基因的克隆、表达及免疫原性研究  

Helicobacter pylori HspA Heat shock Protein Gene: Cloning, Expression and Immunogenicity

作　　者：李明峰[1] 凌贞[1] 张迎春[1] 马爱英[1] 孙建新[2] 施惠娟[2] 吴祥甫[2] 

机构地区：[1]解放军第四五五医院 [2]中国科学院上海生物化学研究所

出　　处：《生物化学与生物物理学报》1999年第3期264-268,共5页

摘　　要：幽门螺杆菌的感染可诱发人体产生胃炎和消化性溃疡，其组成成分热休克蛋白Ａ（ＨｓｐＡ）可刺激机体产生保护性的免疫反应。用ＰＣＲ方法从幽门螺杆菌的染色体ＤＮＡ上扩增出ＨｓｐＡ基因片段，将其插入原核表达载体ｐＥＴ２２ｂ（＋）中，并在ＢＬ２１（ＤＥ３）大肠杆菌表达。经测序ＨｓｐＡ基因片段有３５４ｂｐ组成，可编码１１８个氨基酸残基的多肽。ＳＤＳＰＡＧＥ和免疫印迹分析检测发现，ＨｓｐＡ基因表达的蛋白质分子量约为１５ｋＤ，并证实该重组蛋白质可以被幽门螺杆菌感染阳性患者的血清所识别，同时将其免疫小鼠可刺激机体产生抗该重组蛋白质的抗体。ＨｓｐＡ有可能作为一种有效的蛋白质疫苗用于幽门螺杆菌感染的预防和治疗。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. Hsp A 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.

关 键 词：幽门螺杆菌 热休克蛋白A 克隆 表达 免疫原性 

分 类 号：R378.99[医药卫生—病原生物学] Q513[医药卫生—基础医学]