【摘要】 目的观察刚地弓形虫微线体蛋白6(MIC6)和醛缩酶在虫体内的定位。方法聚合酶链反应扩增微线体蛋白2羧基端(MIC2C)基因片段,构建MIC2C/pGEX-4T-1/BL21表达系统,异丙基-β-D-硫代半乳糖苷诱导表达谷胱苷肽巯基转移酶-MIC2C(GST-MIC2C)蛋白,用该蛋白免疫新西兰兔,制备GST-MIC2C多克隆抗体。刚地弓形虫速殖子涂片,分别用一抗、荧光二抗温育,荧光显微镜下观察。结果成功获得GST-MIC2C多克隆抗体。荧光显微镜下显示MIC6(红色荧光)和醛缩酶(绿色荧光)2种蛋白皆定位于弓形虫的顶端。结论刚地弓形虫MIC6与醛缩酶在虫体内存在共定位关系,为2种蛋白相互作用关系的确立提供了细胞定位学证据。更多还原

【Abstract】 Objective To observe the location of microneme protein 6(MIC6) and aldolase in Toxoplasma gondii.Methods The gene fragment of carboxyl terminus of microneme protein 2(MIC2C) was amplificated by polymerase chain reaction(PCR) and the expression system MIC2C/pGEX-4T-1/BL21 was constructed.The expression of glutathione-S-transferases-MIC2C(GST-MIC2C) protein was induced by isopropyl-β-D-thigalactopyranoside and then this protein was used to immunize New Zealand rabbits to prepare the GST-MIC2C polyclonal antibody.The Toxoplasma gondii were spotted onto slides,and incubated with antibody,immunofluorescence antibody respectively,then were observeded by fluorescent microscope.Results The polyclonal antibody GST-MIC2C was obtained.The fluorescence microscopy displayed that MIC6(red fluorescence) and aldolase(green fluorescence) were positioned at the top of the Toxoplasma gondii.Conclusion MIC6 and aldolase are co-localized in Toxoplasma gondii,which provide the cellular localization evidence for protein-protein interaction.更多还原