【摘要】 以苏云金杆菌 4.0 718菌株杀虫晶体 6 5kD原毒素为材料 ,探索了从SDS -PAGE胶上回收蛋白质进行质谱分析的方法。蛋白纯化的步骤包括SDS -PAGE、电洗脱、脱盐、除SDS。电洗脱采用半透膜法 ,用超滤法脱盐 ,冷丙酮沉淀法除去SDS。结果表明 ,纯化的 6 5kD原毒素经ESI -MS检测 ,分子量约 6 45 0 0Da。经MALDI-MS检测 ,未能有明显蛋白峰出现 ,这可能是该蛋白由于较强的疏水作用 ,溶解性差 ,在与基质混合时处于聚和悬浮态所致。更多还原

【Abstract】 The purpose of the experiment is to use 65 kD protoxin from Bacillus thuringiensis strain 4.0718 insecticidal crystals as a model to explore a way of recovering proteins from SDS-PAGE gel for mass spectrometry. The purifing steps includes SDS-PAGE, electroelution, desalting, and SDS removal. Entrapment of the eluted protein with semipermeable membrane for eletroelution,centrifugation ultrafiltration for desalting and cold acetone method was used for SDS removal.The result shows molecular weight of 65 kD protoxin is about 64500 Da by ESI-MS.There are no obvious protein peaks by MALDI-MS,perhaps soluble nature of the high hydrophic protein is poor, it aggregates in suspension mixing with matrix.更多还原