苏云金杆菌4.0718菌株杀虫晶体65kD原毒素的质谱分析  

Analysis on the 65 kD Protoxin of Bacillus Thuringiensis Strain 4.0718 Insecticidal Crystals for Mass Spectrometry

作　　者：夏立秋 邹先琼[2] 莫湘涛[2] 

机构地区：[1]华中农业大学微生物科学技术系,中国湖北武汉430070 [2]湖南师范大学生命科学学院,中国湖南长沙410081

出　　处：《激光生物学报》2001年第4期281-284,共4页Acta Laser Biology Sinica

基　　金：湖南省青年科技基金项目 (96 - 32 - 0 9);湖南省农业重大攻关项目 (99- 10 0 4- 0 8)

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。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.

关 键 词：苏云金杆菌 原毒素 电洗脱 质谱分析 杀虫蛋白 

分 类 号：Q503[生物学—生物化学] Q939.124