CAJ | 학술논문

目的表达梅毒螺旋体黏附蛋白Tp0751,纯化表达产物并进行免疫活性分析,为探索Tp0751重组蛋白在梅毒致病过程中的作用奠定基础.方法通过生物信息学分析,去除Tp0751信号肽序列,构建原核表达体进行诱导表达;Ni亲和层析柱纯化重组蛋白,Western blot检测其免疫反应性,用重组蛋白免疫新西兰家兔,评价其免疫原性.结果成功构建了pET-28a(+)-0751原核表达载体,经表达、纯化后获得了相对分子质量约为26×103的融合蛋白;Western blot检测其能与梅毒患者阳性血清发生特异性反应;利用纯化的Tp0751重组蛋白免疫新西兰家兔,能诱导家兔产生特异性免疫应答,ELISA法测定免疫血清中特异性抗体滴度在1∶10 240以上.结论重组表达的Tp0751黏附蛋白具有良好的免疫活性,为进一步研究其在梅毒致病过程中的作用和生物学功能奠定了基础.
목적 표체매독라선체점부단백Tp0751,순화표체산물병진행면역활성분석,위탐색Tp0751중조단백재매독치병과정중적작용전정기출.방법 통과생물신식학분석,거제Tp0751신호태서렬,구건원핵표체체진행유도표체;Ni친화층석주순화중조단백,Western blot검측기면역반응성,용중조단백면역신서란가토,평개기면역원성.결과 성공구건료pET-28a(+)-0751원핵표체재체,경표체、순화후획득료상대분자질량약위26×103적융합단백;Western blot검측기능여매독환자양성혈청발생특이성반응;이용순화적Tp0751중조단백면역신서란가토,능유도가토산생특이성면역응답,ELISA법측정면역혈청중특이성항체적도재1∶10 240이상.결론 중조표체적Tp0751점부단백구유량호적면역활성,위진일보연구기재매독치병과정중적작용화생물학공능전정료기출.
Objective To express Tp0751 laminin-binding adhesion of Treponema pallidum (T. pallidum) ,and assess the immunocompetence. Methods The Tp0751 ORF without upstream non-cod-ing region was ligated into the expression vector pET-28a( + ), and expressed in E. coli R2566. Its immuno-gen was analyzed by Western blot and ELISA. Results A fusion protein with molecular weight about 26×103 was attained after expression and purification. Western blot proved that the recombinant protein can specifically react with T. palliclum IgG positive sera. Specific humoral response were elicited after introducing recombinant protein in Zealand rabbit and the specific antibody titer was above 1:10 2400 detected by indi-rect ELISA. Conclusion The expressed recombinant protein showed excellent immunoeompetence, and the results lay the foundation for the research on its function to T. paUidum infection.