CAJ | 학술논문

目的应用分子克隆技术获得纳米级Ag85B蛋白并对其进行鉴定和免疫学特性检测.方法利用S-layer/Streptag I这一具有自我组装能力的纳米模式嵌合结核杆菌分泌的Ag85B蛋白基因,将其克隆到原核表达载体pET28a(+)中,并在大肠杆菌中表达Ag85B蛋白,表达后的蛋白经SDS-PAGE和Western blotting鉴定.结果原核表达重组质粒pET28a-Streptag I-SbpA-Ag85B成功构建,并可在大肠杆菌中诱导表达,得到的Streptag I-Sbpa-Ag85B蛋白经SDS-PAGE和Western blotting鉴定正确,重组蛋白能被结核患者血清识别.结论重组后的Ag85B蛋白具有较强的免疫反应性.所建立的原核表达体系为新型结核疫苗研制提供了一个新的思路.
목적 응용분자극륭기술획득납미급Ag85B단백병대기진행감정화면역학특성검측.방법 이용S-layer/Streptag I저일구유자아조장능력적납미모식감합결핵간균분비적Ag85B단백기인,장기극륭도원핵표체재체pET28a(+)중,병재대장간균중표체Ag85B단백,표체후적단백경SDS-PAGE화Western blotting감정.결과 원핵표체중조질립pET28a-Streptag I-SbpA-Ag85B성공구건,병가재대장간균중유도표체,득도적Streptag I-Sbpa-Ag85B단백경SDS-PAGE화Western blotting감정정학,중조단백능피결핵환자혈청식별.결론 중조후적Ag85B단백구유교강적면역반응성.소건립적원핵표체체계위신형결핵역묘연제제공료일개신적사로.
Objective To obtain nanometer Ag85B protein by the molecular cloning techniques and identify its immunological characteristics.Methods The gene sllB coding surface layer protein (S-layer)was truncated and linked with the Mycobacterium tuberculosis secreted protein Ag85B gene.The expression plasmid pET28a-Streptag I-SbpA-Ag85B was transformed into E.coli,the protein was identified by SDS-PAGE and Western blotting.Results The expression system was developed successfully for recombinant nanometer-scale antigen and the fusion protein was identified by SDS-PAGE with correct molecular weight and can be recognized by the serum of tuberculosis patients.Conclusion The recombinant Ag85 B protein has strong immune reaction and small size on nanometer level,this expression system provides a new way for the tuberculosis vaccine design.