摘要
描述了一种改良的高效细菌感受态的制备和感受态细胞的保藏及细菌的转化方法.三种不同的大肠杆菌分别是DH5α,TG1和XL1blue,其中最有效的是XL1blue.每种菌株的感受态细胞制备的最佳光密度(OD600值)不同.对感受态细胞的存储时间及其与转化效率之间的关系也进行了研究,结果显示感受态细胞可以在-20℃存储7d,在-70℃存储15d.将常见的方法做了三种改进:首先是将CaCl2溶液改为TB溶液;其次是将培养基由LB换成S.O.C;再就是在质粒对感受态细胞的转化过程中加入DMSO或PEG8000.改良的细菌转化系统比常见的方法能提高转化效率约1000倍,是一种高效的细菌转化系统.
An efficient bacterial transformation system was described for the preparation of competent cells, plasmid preparation, and for the storage in bacterial stocks. Three different strains of Escherichia coli that were tested were DHSa, TG1 and XL1 blue, and the most efficient strain being XL1 blue. The optimal optical density （OD600） range for competent cell preparation varied for each of the strains. The storage time of competent cells and its correlation to transformation efficiency were studied. Three critical alterations to previous methods were made, which were the changing of the normal CaCl2 solution to TB solution, the changing of the medium from LB to S. O. C, and addition of DMSO or PEG8000 during transformation. The transformation efficiency was increased by about 1 000 times by the use of this improved bacterial transformation system.
出处
《华中科技大学学报（自然科学版）》
EI
CAS
CSCD
北大核心
2006年第4期112-115,共4页
Journal of Huazhong University of Science and Technology(Natural Science Edition)
基金
国家高技术研究发展计划资助项目(2002AA224031)
华中科技大学博士论文基金资助项目.
关键词
感受态细胞
大肠杆菌
质粒
保藏
转化
competent cells
Escherichia coli （E. coli）
plasmid
storage
transformation
