目的 筛选与弓形虫微线体蛋白6羧基端(MIC6C)相互作用的蛋白.方法 以GST-MIC6C蛋白作为探针蛋白,利用GST pull-down技术,从弓形虫裂解液中筛选与其相互作用的蛋白,SDS-PAGE分析实验产物；将目标蛋白条带转印至PVDF膜,测蛋白序列,BLAST2在线对比搜索相似蛋白序列,初步确定目标蛋白.制备目标蛋白的抗体,Western blot分析GST pull-down实验产物.结果 GST pull-down实验筛选到的目标蛋白的序列与弓形虫醛缩酶的序列一致；制备了醛缩酶的多克隆抗体,目标蛋白条带能被该抗体特异地识别.结论 采用GST pull-down技术,初步筛选出与弓形虫MIC6C相互作用的蛋白为醛缩酶.%To screen the interacting protein from tachyzoite lysates in C terminal of microneme protein 6 (MIC6C) of Toxoplasma gondii, glutathione sepharose beads were incubated with GST-MIC6C protein,and then incubated with tachyzoite lysates. Bound proteins were eluted by using sample buffer, and eluants were resolved by SDS-PAGE and transferred onto pol-yvinylidene difluoride (PVDF) membranes. Protein band were cut from PVDF membrane, and the protein were subjected to N-terminal amino acid sequencing. BLAST2 server in Internet was used to analyse the amino acid sequences. Resutts showed that the N-terminal amino acid sequence of target protein was coincident with the amino acid sequence of aldolase, and the target protein band on NC membrane was specifically recognized by anti-aldolase antibody. The protein which interacts with C terminal of microneme protein 6 (MIC6C) of Toxoplasma gondii is aldolase.
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