一种简捷有效的大肠杆菌感受态细胞制备方法  

A Simple and Effective Method for Preparation of Escherichia coli Competent Cells

作　　者：代红星[1] 张庆桥[2] 樊宝良[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071000 [2]河北工程大学农学院,河北邯郸056038

出　　处：《安徽农业科学》2008年第29期12627-12628,共2页Journal of Anhui Agricultural Sciences

摘　　要：[目的]探索建立简捷快速有效的大肠杆菌感受态细胞制备方法。[方法]在已有的TSS法基础上进行了进一步的改进，建立了一个只需一步室温离心便能够获得与常规方法效价相近，能够长期保存的大肠杆菌感受态细胞的制备方法。[结果]新方法制备的感受态细胞每微克转化菌落数达105-106个，可以满足在质粒中进行的常规克隆需要。不同冻存时间对新方法制备的感受态细胞转化效率无显著影响。新方法制备的感受态细胞可以立即使用也可以于-80℃下长期保存至少9个月不会影响其转化效价。这也与常规CaC l2法制备的感受态细胞性能相当。[结论]该方法为一次大量制备并长期保存使用感受态细胞奠定了基础。[ Objective ] The aim was to explore the simple and effective method for preparation of Escherichia coli competent cells. [ Method ] A method for preparation of E. coli competent cells was constructed by the further improvement on the basis of the known TSS method, which needed only one step of centrifugation at room temperature to obtain the E. coli competent cells preserved in a long time and had the similar efficiency as the routine method. [ Resuhl The number of per μg colony transformed by the competent cells prepared by the new method reached 105 - 106 , which could meet the demand of routine clone completed in the plasmid. Different cryopreservation time had no significant influence on the transformation efficiency of the competent ceils prepared by the new method, and the ceils could be used immediately and also could be preserved in a long time at - 80 ℃, whose transformation efficiency could not be influenced at least 9 months and its performance was equivalent to that prepared by the routine CaCI2 method. [ Conclusion ] The method laid the foundation for large scale preparation of competent ceils in one time, and preservation and usage of competent ceils in a long time.

关 键 词：大肠杆菌 感受态细胞 CaCl2法 制备 

分 类 号：S182[农业科学—农业基础科学]