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  

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

作　　者：宋晟[1] 夏立秋[1] 黄江丽[1] 孙运军[1] 丁学知[1] 

机构地区：[1]湖南师范大学生命科学学院湖南省高校微生物学重点实验室,长沙410081

出　　处：《微生物学报》2005年第3期467-471,共5页Acta Microbiologica Sinica

基　　金：国家自然科学基金 ( 3 0 2 70 0 3 7;3 0 470 0 43 );国家"863计划"( 2 0 0 2AA2 45 0 2 1);湖南省自然科学基金重点项目( 0 4JJ2 0 0 3 );湖南省教育厅重点项目 ( 0 2C0 17)~~

摘　　要：根据苏云金杆菌4 0 718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 assistedlaserdesorption ionizationtimeofflightmassspectrometry ,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 0 718菌株伴孢晶体中所含的Cry1Ac和Cry2Aa蛋白,其精确分子量分别为13416 0Da和710 97Da。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.

关 键 词：苏云金杆菌 蛋白质组 杀虫晶体蛋白 双向电泳 基质辅助激光解吸电离飞行时间质谱 

分 类 号：S482.3[农业科学—农药学]