一种纯化苏云金杆菌Cry1Ac蛋白的方法  

An Improved Method for Purifying Cry1Ac Protein from Bacillus thuringiensis

作　　者：王发祥[1,2] 刘永乐[1] 丁学知[2] 夏立秋[2] 

机构地区：[1]长沙理工大学化学与生物工程学院,长沙410114 [2]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,长沙410081

出　　处：《湖南农业科学》2011年第8期117-119,共3页Hunan Agricultural Sciences

基　　金：国家"863计划"(2008AA100801);湖南省科技支撑计划(2010NK3003);湖南省教育厅项目(10C0400)

摘　　要：Cry蛋白的纯化技术在很大程度上制约了其结构与功能的研究。为此,提出了一种以等电点沉淀方法纯化苏云金杆菌Cry1Ac的原毒素,用分子筛层析纯化其活性毒素的方法。结果表明:Cry1Ac的原毒素在pH值5.05条件下沉淀纯化效果好;以纯化的原毒素激活进行分子筛层析,收集其最大峰便可获得较高浓度的活性毒素;该方法不仅纯化效果好,而且省去层析后的浓缩步骤,是Cry蛋白的纯化方法研究中的一次革新,对推动Cry蛋白的结构和功能研究具有重要的意义。The research of structure and function of Cry proteins was greatly restricted by their purification method. Therefore, a method that purifying Bacillus thuringiensis CrylAc protoxin by using isoeleetric precipitation and purifying Bacillus thuringiensis CrylAc activated toxin by using molecular sieve chromatography was proposed. The results showed that the CrylAc protnxin had good effects of precipitation and purification when pH value was 5.05; activating the purified protoxin to conduct molecular sieve chromatography, high concentration of CrylAc activated toxin could be obtained from the maximum peak of chromatography. This improved method not only has good purification effect, but also contribute a renovation to the purification method of Cry protein, which is significant for promoting researches on structure and function of Cry protein.

关 键 词：苏云金杆菌 CRY1AC 纯化 等电点沉淀 分子筛层析 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]