Gunn L, Nickoloff J A
Department of Cancer Biology, Harvard School of Public Health, Boston, MA 02115, USA.
Mol Biotechnol. 1995 Apr;3(2):79-84. doi: 10.1007/BF02789103.
A simple and reproducible method for transferring low copy-number episomal plasmids from yeast to Escherichia coli has been developed. Although slightly more time-consuming than direct transfer methods, which are effective with high copy number plasmids, the method is significantly faster than methods that require purification of yeast DNA. Plasmid DNA is released from yeast cells during brief treatments involving grinding with glass beads and heating. The treated yeast are cooled, electrocompetent E. coli is added, the mixture is electroporated, and transformants are selected using standard conditions for E. coli electrotransformation. The procedure typically yields sufficient transformants for most applications.
已开发出一种将低拷贝数附加体质粒从酵母转移至大肠杆菌的简单且可重复的方法。尽管该方法比直接转移方法耗时稍长（直接转移方法对高拷贝数质粒有效），但比需要纯化酵母DNA的方法要快得多。在涉及用玻璃珠研磨和加热的短暂处理过程中，质粒DNA从酵母细胞中释放出来。将处理后的酵母冷却，加入电感受态大肠杆菌，对混合物进行电穿孔处理，然后使用大肠杆菌电转化的标准条件筛选转化体。该程序通常能产生足够的转化体以满足大多数应用需求。
