梅毒螺旋体Tp0751重组蛋白的表达及鉴定  

Expression and Identification of Tp0751 Recombinant Protein from Treponema Pallidum

作　　者：肖勇健[1] 张秋桂[2] 刘双全[2] 谭湘芳[1] 赵飞俊[3] 

机构地区：[1]南华大学第二附属医院,湖南衡阳421001 [2]南华大学第一附属医院 [3]南华大学病原生物学研究所

出　　处：《南华大学学报（医学版）》2009年第3期264-266,共3页Journal of Nanhua University(Medical Edition)

基　　金：湖南省卫生厅资助课题(B2006-115);湖南省教育厅资助课题(07C626)

摘　　要：目的表达梅毒螺旋体粘附蛋白Tp 0751,纯化表达产物并对其抗原性进行鉴定。方法通过生物信息学分析,去除Tp 0751信号肽序列,构建原核表达体并在大肠埃希菌(E.coli)R2566中进行诱导表达;Ni-NTA法纯化重组蛋白;蛋白质印迹法(WB)鉴定重组蛋白。结果成功构建了PET-28a(+)/Tp 0751原核表达载体;经表达、纯化后获得了相对分子量约为26 kD的融合蛋白;经WB分析能与梅毒患者阳性血清发生特异性反应。结论重组表达的Tp 0751重组蛋白具有良好的抗原性,为进一步研究Tp 0751重组蛋白在梅毒致病过程中的作用奠定了基础。Objective To express Tp 0751 laminin - binding adhesin from Treponema pallidum （ T. pallidum ） and analyse the antigenicity of recombinant protein. Methods The Tp 0751 DNA fragment without upstream non - coding region was ligased into the expression vector PET - 28a （ ＋ ） , and expressed in Escherichia coli （ E. coli） R2566. The recombinant protein was purified with Ni - NTA affinity chromatography, and analyzed by SDS - PAGE and western blot. Results Prokaryotic expression vector PET -28a（ ＋ ）/Tp 0751 was constructed successfully and a fusion protein with molecular weight about 26kD was attained. Western blot showed that the recombinant protein could specifically react with T. pallidum IgG positive sera. Conclusion The expressed recombinant protein showed better antigenicity, and the results lay the foundation for the research on its pathogenic function.

关 键 词：梅毒螺旋体 粘附蛋白 Tp0751 

分 类 号：R759.1[医药卫生—皮肤病学与性病学]