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目的表达和纯化弓形虫表面抗原1(SAG1),并分析其免疫学活性。方法构建pET15b-SAG1质粒,转化到宿主菌E.coil BL21(DE3)中,经IPTG诱导表达,使用Ni 2+亲和层析柱对重组蛋白进行纯化,用间接ELISA检测目的蛋白的抗原性。结果 SDS-PAGE显示表达的SAG1蛋白为可溶性的,经纯化后其纯度可达到90%以上;ELISA检测表达蛋白能被弓形虫感染者血清识别;棋盘滴定显示重组SAG1蛋白适宜包被浓度为0.5μg/ml。结论本研究表达的SAG1蛋白为可溶性的,且具有良好的抗原性,为进一步建立弓形虫血清学诊断方法和研制弓形虫病疫苗奠定了基础。
Objective To express and purify Toxoplasma gondii surface antigen 1 (SAG1) and analyze its immunological activity. Methods The recombinant plasmid pET15b-SAG1 was transformed into E. coli BL21 (DE3) and induced by IPTG. The recombinant protein was purified by Ni2 + affinity chromatography. The antigenicity of the recombinant protein was detected by indirect ELISA. Results SDS-PAGE showed that the expressed SAG1 protein was soluble, and its purity was over 90% after purification. The protein detected by ELISA could be recognized by the serum of Toxoplasma gondii infection. The chemostat titration showed that the recombinant SAG1 protein was suitable for coating 0.5μg / ml. Conclusion The SAG1 protein expressed in this study is soluble and has good antigenicity, which lays the foundation for the further establishment of a serological diagnostic method for Toxoplasma gondii and the development of a vaccine against toxoplasmosis.