弓形虫复合抗原基因SAG1/ROP1在大肠杆菌中表达的初步研究

EXPRESSION OF TOXOPLASMA GONDII MULTI-ANTIGENIC GENE ENCODING SAG1 AND ROP1 IN ECOLI

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 pET2 8aRT7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pET2 8-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 6 6kD ,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。

Aim To construct a recombinant plasmid encoding Toxoplasma gondii multi-antigen SAG1/ROP1 and to detect the immunity of the expressed product in E coli.Methods Subclone the SAG1 and ROP1 genes into the downstream of the T7 promoter of vector pET28a respectively,transform the recombinant plasmid into competent cells of E coli DH5α To assess the potential immunity of the recombinant expressed product,the expressed product,induced by IPTG,was analysed with SDS-PAGE and Western-blot.Results The recombinant plasmid pET28a-SAG1/ROP1.SDS-PAGE and Western-blotting indicated the molecular weight of expressed product was about 66kDa and had immunoactivity.Conclusions The recombinant plasmid coding for Toxoplasma gondii SAG1 and ROP1 can express in E coli DH5α and the expressed product had immmunoactivity which mayb uses as the candidate vaccine for Toxoplasosis.