苏云金芽孢杆菌以色列亚种毒素蛋白定量检测方法分析  

A Toxin Protein Quantitative Detection Method for Bacillus Thuringrensis Subsp. Israelensis

作　　者：金海燕 胡凌云 刘荷梅 周莉 胡虓 程治国 李青 刘华梅 JIN Hai-yan;HU Ling-yun;LIU He-mei;ZHOU Li;HU Xiao;CHENG Zhi-guo;LI Qing;LIU Hua-mei(Wuhan Kernel Bio-tech Co.Ltd,Wuhan 430074,China)

机构地区：[1]武汉科诺生物科技股份有限公司,武汉430074

出　　处：《湖北农业科学》2019年第3期96-101,共6页Hubei Agricultural Sciences

摘　　要：为完善苏云金芽孢杆菌以色列亚种(Bacillus thuringrensis subsp. israelensis)毒素蛋白定量检测方法,试验用一定浓度的磷酸缓冲溶液、纯化水洗涤苏云金芽孢杆菌以色列亚种试样,去除培养基残留。用SDS-PAGE电泳法检测离心后收集到的试样菌体毒素蛋白含量。结果表明,通过磷酸缓冲液及水洗涤试样,解决了试样中杂质干扰毒素蛋白含量检测的关键性问题,采用该方法能够准确对苏云金芽孢杆菌以色列亚种两种主要相对分子质量的毒素蛋白进行检测与定量,且重复性好,检测误差可以控制在5%以内,并且检测结果与产品生物效价存在显著相关性。To provide a quantitative detection method for toxin protein of Bacillus thuringrensis subsp.israelensis(Bti),and to improve the quality control technique.The samples of Bacillus thuringrensis subsp.israelensis were washed in water with a certain concentration of phosphoric acid buffer solution,and the residue of culture medium was removed.The bacterial toxin protein collected after centrifugation was detected by SDS-PAGE electrophoresis and the content was determined.The key problem of the determination of impurity interfering toxin protein in the sample was solved by washing the sample with phosphoric acid buffer and water.This method can accurately detect and quantify the two major molecular weight toxin proteins of Bacillus thuringrensis subsp.israelensis.The method was used to detect the subspecies of Bacillus thuringrensis subsp.israelensis.The results were reproducible,the detection error could be controlled within 5%of the average value,and there was a significant correlation between the detection results and the biological titer of the products.The method is reasonable in conception and simple in operation,and can provide reference for further product quality management.

关 键 词：苏云金芽孢杆菌以色列亚种(Bacillus thuringrensis subsp.israelensis) 毒素蛋白 SDS-PAGE凝胶 电泳 

分 类 号：TQ453.5[化学工程—农药化工] O657.8[理学—分析化学]