苏云金杆菌4.0718菌株杀虫晶体蛋白的双向电泳飞行时间质谱分析

根据苏云金杆菌4.0718菌株杀虫晶体蛋白的特性，对裂解液组成、上样量、聚焦时间等相关技术进行了比较研究和条件优化，首次获得苏云金杆菌杀虫晶体蛋白双向电泳图谱，并对部分蛋白质点进行胰酶酶解，基质辅助激光解吸电离飞行时间质谱（matrixassisted laser desorption/ionization time of flight mass spectrometry，MALDITOFMS）测定肽质量指纹图谱，Mascot软件查询SwissProt数据库，最终鉴定出苏云金杆菌4.0718菌株伴孢晶体中所含的Cry1Ac和Cry2Aa蛋白,其精确分子量分别为134160Da和71097Da。

Analysis of insecticidal crystal proteins from Bacillus thuringiensis strain 4.0718 through two-dimensional gel electrophoresis and MALDI-TOF-Mass spectrometry

The present study analyses the insecticidal crystal proteins(ICPs) fromBacillus thuringiensis strain 4.0718 through two-dimensional electrophoresis and matrix-assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF-MS). By comparing and optimizing the composition of lysis solution, the volume of sample loading and the protocol for isoelectric focusing, a well-focused 2-DE map with high resolution and reproducibility was achieved for the first time.And after an tryptic enzymolisis and a test of part of protein spots by means of MALDI-TOF-MS, the peptides mass fingerprint(PMF) was obtained and, by referring to Swiss-Prot, Cry1Ac and Cry2Aa contained in Bt 4.0718 were identified ,with their molecular weights 134160 Da and 71097 Da respectively.