苏云金芽孢杆菌伴胞晶体蛋白提取方法比较研究  

A Comparison with Different Methods of Extraction of Crystal Protein form Bacillus thuringiensis

作　　者：罗朝辉[1] 夏立秋[1] 丁学知[1] 王海龙[1] 曾智[1] 

机构地区：[1]湖南师范大学生命科学学院微生物分子生物学湖南省重点实验室,中国长沙410081

出　　处：《湖南师范大学自然科学学报》2007年第3期94-98,共5页Journal of Natural Science of Hunan Normal University

基　　金：国家自然科学基金资助项目(30670052);国家863计划资助项目(2006AA02Z187;2006AA10A212);湖南省自然科学基金重点资助项目(06jj2009)

摘　　要：采用液体双相分层法和等电点沉淀法提取苏云金芽孢杆菌伴胞晶体蛋白．原子力显微镜观测，发现液体双相分层提取的晶体蛋白具有完整典型的晶体结构；等电点法提取的得率高，不能检测到完整的晶体结构．聚丙烯酰胺凝胶电泳分析，等电点法提取的蛋白质含量是液体双相法提取的蛋白含量的1～2倍．将2种方法提取的晶体蛋白对初孵棉铃虫幼虫生物测定均表现杀虫生物活性，经统计分析，液体双相法提取的蛋白质生测毒力比等电点法的高1．5～3倍．It is reported that a comparison with the different of extracting parasporal Crystal Protein of Bacillus thuringiensis between the. liquid double-phase method and Isoelectric point method. Using atomic force scan （AFM） electron microscope as stool, It is found that proteins extracted by liquid double-phase method have the typical shape and structure,while that of extracted by isoelectric point method have no typical crystal shape. However, when analyzed by SDS-PAGE, the result show that pure protein content extracted by the Liquid double-phase method is 1 - 2 times less than that of extracted by isoelectric point method. The bioassay is performed on proteins extracted by two different methods. The result shows that proteins extracted by either of the two methods work effectively on young instar larvae of Helicouerpa armigera. But statistic analysis show that proteins extracted by liquid double-phase was 1.5 - 3 times more toxic than that of extracted by isoelectric point.

关 键 词：苏云金芽孢杆菌 液体双相分层 等电点 伴胞晶体蛋白 原子力显微镜 聚丙烯酰胺凝胶电泳 

分 类 号：Q936[生物学—微生物学]