幽门螺杆菌黏附素基因（hpaA）的表达及重组蛋白的免疫原性检测  

Expression of adhesion gene of Helicobacter pylori and immunogenicity determination of expressed product

作　　者：杨芸 孔超[2] 徐帆洪[2] 李笑梅[1] 

机构地区：[1]上海健康职业技术学院生物医药系,200237 [2]上海生物制品研究所有限责任公司第一研究室,200052

出　　处：《国际生物制品学杂志》2015年第1期10-12,共3页International Journal of Biologicals

摘　　要：目的 研究幽门螺杆菌黏附素A（adhesin A of Helicobacter pylori,HpaA）基因在大肠杆菌中的表达和检测纯化HpaA的免疫原性.方法 将hpaA/pET28a表达质粒转入大肠杆菌BL21（RP）株并用异丙基-β-D-硫代半乳糖苷诱导表达,通过Ni2＋亲和层析和分子筛层析纯化表达蛋白.用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表达蛋白的相对分子质量,用蛋白质印迹法对表达蛋白进行特性确认.将纯化HpaA腹腔注射BALB/c小鼠,并检测小鼠的抗体水平.结果 表达蛋白的相对分子质量约为30 000,与HpaA相符.纯化的表达蛋白可与抗HpaA抗体发生特异性反应.腹腔注射HpaA的小鼠与对照小鼠间的血清抗HpaA IgG抗体水平差异存在统计学意义（t=8.367,P＜0.01）.结论 HpaA在大肠杆菌BL21（RP）株中得到成功表达,且纯化HpaA具有较好的免疫原性.Objective To study the expression of adhesin A of Helicobacter pylori （hpaA） in E.coli and determine immunogenicity of purified HpaA.Methods hpaA/pET28a expression plasmid was transmitted into E.coli strain BL21 （RP） and HpaA expression was induced by IPTG.The expressed product was purified by Ni2＋ affinity chromatography and molecular sieve chromatography.Relative molecular mass of expressed product was determined by SDS-PAGE and characteristics of expressed product was identified by Western blotting.BALB/c mice were immunized with purified HpaA by intraperitoneal injection,and antibody levels in immunized mice were determined.Results Relative molecular mass of the expressed product was 30 000,consistent with that of HpaA.The purifed expression product specifically reacted with anti-HpaA antibody.The levels of serum anti-HpaA IgG antibodies had significant difference between mice immunized with purified HpaA by intraperitoneal injection and control mice （t=8.367,P〈0.01）.Conclusion HapA is successfully expressed in E.coli strain BL21 （RP）,and purified HpaA has good immunogenicity.

关 键 词：螺杆菌 幽门 黏附素 细菌 基因表达 免疫原性 

分 类 号：R378.99[医药卫生—病原生物学]