弓形虫复合抗原基因SAG1/ROP1在大肠杆菌中rn表达的初步研究

摘要

Aim To construct a recombinant plasmid encoding Toxoplasma gondii multi- antigen SAG1/ROP1 and to detect rnthe immunity of the expressed product in E coli. Methods Subclone the SAG1 and ROP1 genes into the downstream of the T7 prornmoter of vector pET28a respectively, transform the recombinant plasmid into competent cells of E coli DH5α. To assess the potential rnimmunity of the recombinant expressed product, the expressed product, induced by IPTG, was analysed with SDS - PAGE and Westrnem - blot. Results The recombinant plasmid pET28a - SAG1/ROP1. SDS - PAGE and Western - blotting indicated the molecular rnweight of expressed product was about 66kDa and had immunoactivity. Conclusions The recombinant plasmid coding for Toxoplasrnma gondii SAG1 and ROP1 can express in E coli DH5a and the expressed product had immmunoactivity which mayb uses as the canrndidate vaccine for Toxoplasosis.%目的 构建含弓形虫主要表面抗原基因SAG1和棒状体蛋白ROP1复合基因重组质粒，并在大肠杆菌中表达，检测复合基因表达产物的免疫活性。方法用亚克隆技术分别把SAG1与ROP1基因克隆至pET28aR T7启动子下游，转化大肠杆菌DH5α感受态细胞，经IPTG诱导表达，SDS-PAGE及Western-Blot分析。结果获得pET28-SAG1/ROP1重组表达载体；SDS-PAGE及Western-Blot印迹显示SAG1/ROP1复合基因表达蛋白产物分子量约为66kD，具有一定的免疫活性。结论弓形虫复合抗原基因SAG1/ROP1可在大肠杆菌中表达，表达产物具有免疫活性。