CAJ | 학술논문

目的表达E型沙眼衣原体主要外膜蛋白(MOMP)并制备其兔源多克隆抗体.方法诱导表达实验室构建的MOMP/pGEX6p-1重组质粒,通过胶回收进行目的蛋白的纯化.MOMP免疫新西兰家兔制备多克隆抗体,ELISA法测定抗体效价,Western印迹、细胞免疫荧光方法鉴定抗体的特异性.结果在大肠埃希菌中成功表达融合蛋白谷胱甘肽巯基转移酶(GST)-MOMP.ELISA法检测抗体的效价达到1∶12 800,Western印迹结果显示抗体可与原核表达的MOMP特异性结合,细胞免疫荧光检测结果显示抗体可与体外培养的沙眼衣原体特异性结合.结论成功表达了E型沙眼衣原体的MOMP,并制备了高效价、高特异性的抗MOMP抗体,为沙眼衣原体的检测和临床研究提供依据.
목적 표체E형사안의원체주요외막단백(MOMP)병제비기토원다극륭항체.방법 유도표체실험실구건적MOMP/pGEX6p-1중조질립,통과효회수진행목적단백적순화.MOMP면역신서란가토제비다극륭항체,ELISA법측정항체효개,Western인적、세포면역형광방법감정항체적특이성.결과 재대장애희균중성공표체융합단백곡광감태구기전이매(GST)-MOMP.ELISA법검측항체적효개체도1∶12 800,Western인적결과현시항체가여원핵표체적MOMP특이성결합,세포면역형광검측결과현시항체가여체외배양적사안의원체특이성결합.결론 성공표체료E형사안의원체적MOMP,병제비료고효개、고특이성적항MOMP항체,위사안의원체적검측화림상연구제공의거.
Objective To express major outer membrane protein (MOMP) of Chlamydia trachomatis E and to prepare rabbit polyclonal antibody. Methods The recombinant plasmid MOMP/ pGEX6p-l prepared by our lab was introduced into E. coli. The protein was expressed and purified by gel recycling, then was injected into New Zealand rabbits to produce polyclonal antibodies. Enzyme linked immunosorbent assay (ELISA) was used to detect the titer of antibody. The antibody specificity was identified by Western blot and immunofluorescence. Results The fusion recombinant protein glutathione S-transferase (GST)-MOMP was successfully expressed in E. coli. The titer of antibody recombinant protein detected by Western blot and to the endogenous MOMP of Chlamydia trachomatis in vitro detected by immunofluorescence. Conclusions The recombinant MOMP is successfully expressed and the MOMP antibody with high titer and high specificity is obtained. which will be helpful for Chlamydia trachomatis detection and related clinical research.