弓形虫主要表面抗原1截短型片段的纯化与鉴定

目的:表达和纯化弓形虫SAG1,分析其免疫原性。方法:将诱导表达SAG1后菌体裂解,离心,分别收集其上清和沉淀;亲和层析分离纯化裂解上清液中的SAG1,SDS-PAGE和Western blot鉴定SAG1纯度和免疫原性。结果:诱导后重组体pGEX-SAG1/Bl21超声裂解上清液、8M脲溶解沉淀的上清中都含有融合蛋白GST-SAG1,从超声裂解上清液中纯化获得融合蛋白GST-SAG1。结论:亲和层析获得具有免疫原性的SAG1。

Purification and Identification of Toxoplasma gondii Major Surface Antigen 1 Truncated Fragment

Objective To express and purification the major surface antigen 1 truncated fragment of Toxoplasma gondii and analyse its immunogenicity.Methods The recombinant clone pGEX-SAG1/Bl21 was induced with IPTG to express target protein SAG1,and expressed recombinant protein was purified by affinity chromatography;The purifity and immunogenicity of this protein was analyzed by SDS-PAGE and western-blot reacting with the sera from rabbits infecting with Toxoplasma gondii.Results the major surface antigen 1 both existed in the supernatant and the precipitate,recombinant protein was purified by affinity chromatography from the supernatant of pGEX-SAG1/Bl21 lysate;The purified protein reacted with the rabbit sera infected with Toxoplasma gondii.Conclusion The major surface antigen 1 with good immunogenicity was purified from the supernatant of pGEX-SAG1/Bl21 lysate.