一种改良的恶性疟原虫核蛋白和胞浆蛋白分离方法的建立

发布时间：2018-07-28 08:44

【摘要】：目的建立一种改良的恶性疟原虫核蛋白和胞浆蛋白分离方法,并对分离的蛋白组分进行分析和鉴定。方法分别用含不同盐浓度的裂解缓冲液和不同研磨次数提取核蛋白和胞浆蛋白,并利用针对不同组分特异性蛋白的抗体进行Western blot,分析分离效果。结果含50mmol/L NaCl的裂解缓冲液和研磨100~150次为分离核蛋白和胞浆蛋白最佳条件,恶性疟原虫醛缩酶胞浆蛋白和组蛋白3核蛋白的抗体Western blot、抗GFP抗体Western blot及活细胞免疫荧光检测均表明该方法效果良好。结论建立了一种快速、有效的恶性疟原虫核蛋白和胞浆蛋白的分离方法(裂解液盐浓度为50mmol/L,研磨100~150次),为研究恶性疟原虫蛋白的胞内分布以及蛋白质/蛋白质/核酸相互作用提供了重要的技术手段。
[Abstract]:Objective to establish an improved method for the isolation of nucleoprotein and cytosolic protein from Plasmodium falciparum, and to analyze and identify the components of the proteins isolated from Plasmodium falciparum. Methods the nucleoprotein and cytosolic protein were extracted with different salt concentration and different grinding times respectively. Western blots were used to analyze the separation effect of the antibodies against different component specific proteins. Results the optimal conditions for the separation of nuclear and cytoplasmic proteins were the cleavage buffer containing 50mmol/L NaCl and 100m lapping. The antibodies of plasmodium falciparum aldolase cytosolic protein and histone 3 nucleoprotein were detected by Western blot, anti GFP antibody Western blot and living cell immunofluorescence. Conclusion A fast method is established. The effective isolation method of plasmodium falciparum nucleoprotein and cytosolic protein (50 mmol / L lysate salt concentration, 150 grinding times) provides an important technical means for studying the intracellular distribution of Plasmodium falciparum protein and the interaction between protein and nucleic acid.
【作者单位】：同济大学传染病与疫苗研究所;
【基金】：国家自然科学基金项目(No.31271388) 中央高校基本科研业务费专项资金
【分类号】：R382.31

【共引文献】