一种优化的大肠杆菌感受态细胞制备及转化方法  

An Optimized Method for Preparation and Transformation of Escherichia coli Competent Cells

作　　者：黄学娟 张金迪 张壮 李裕静 曹雪松[1] 胡全安 

机构地区：[1]聊城大学生命科学学院,聊城252000

出　　处：《基因组学与应用生物学》2017年第12期5199-5204,共6页Genomics and Applied Biology

基　　金：山东省自然科学基金面上项目(编号:ZR2014CM001)资助

摘　　要：Silwet L-77是一种非离子型的表面活性剂,常用于植物的转化。本研究发现,Silwet L-77的加入也可以显著地提高大肠杆菌的转化效率。同时,我们比较了不同培养温度、不同培养浓度(OD_(600)值)及不同冷冻保护剂对感受态细胞转化效率的影响。我们发现,28℃培养E.coli至OD_(600)值为0.55~0.6之间时制备感受态细胞,利用9%的DMSO做为冷冻保护剂冷冻保存感受态细胞,转化时加入0.001 5%~0.002%的Silwet L-77,可以获得最高的转化效率。总之,该研究进一步优化了大肠杆菌感受态细胞的制备及转化方法。Silwet L-77 is a nonionic surfactant which is widely used in plant transformation.We found that applying the surfactant also enhances the E.coli transformation efficiency.Meanwhile,we compared the cultivation temperature,the cultivation concentration of the E.coli,as well as the effect of different freezing protection agent on transformation efficiency of competent cells.The E.coli growed at 28℃ till the culture density OD600reached about 0.550.6 when the competent cells were prepared stored with 9% DMSO in deep freezer.In the end,add 0.001 5%0.002% Silwet L-77 to the transformation mixture in order to acquire the highest efficiency.In summary,the research might provide an optimized method for preparation and transformation of E.coli competent cells.

关 键 词：大肠杆菌 感受态细胞 质粒 转化 

分 类 号：Q93[生物学—微生物学]