摘要
目的:优化外源性DNA导入XL1-blue菌株电转化条件,摸索构建噬菌体抗体库构建过程较优化的电转化条件。方法:培养XL1-blue菌株,在不同培养时间制作感受态细胞,应用不同电压分别作电转化,并对其转化效果进行鉴定,确定较好的培养时间和电压后,再确定较好的电击时间。结果:不同条件的电转化对于抗体库构建的影响有较大的差别。结论:感受态细胞数量在OD600nm0.5～0.8、电压为2.0kV、电击时间为3ms时,电转化能达到更好的效果。
Objective： To explore the optimal conditions for electrotransformation of exogenous DNA into XL1 - blue strain. Methods： XL1 - blue strain was cultured to make competent cells with various OD600nm values at different times, respectively, and then exogenous DNA were electrotransformed into these ceils at 1.0,1.5,2.0 and 2.5 kV. The optimized voltage and number of competent ceils were evaluated according to The effects of different efficiency and then the optimized lightning stroke time was determined based on above two optimized conditions. Results ： conditions on antibody library construction were very different. Conclusion： The optimal electrotransformation effect can be obtained when OD600 nm is between 0.5 and 0.8, voltage is 2.0 kV and lightning stroke time is 3 millisecond.
出处
《西南国防医药》
CAS
2009年第5期507-508,共2页
Medical Journal of National Defending Forces in Southwest China
关键词
电转化
抗体库
感受态细胞
electrotransformation
antibody library
competent cell
