Sinclair M I, Morgan A F
Aust J Biol Sci. 1978 Dec;31(6):679-88. doi: 10.1071/bi9780679.
A procedure has been developed which allows transformation of P. aeruginosa strain PAO with plasmid and bacteriophage DNA at a frequency of 10(-6) per recipient cell. The method is similar in outline to that developed for Escherichia coli. It involves growing the recipient cells to 3-5 x 10(8) per ml in nutrient broth, washing the cells with 0.1 M MgCl2, resuspending in 0.175 M CaCl2 for 20 min, exposing to DNA for 1 h and then heat pulsing at 42 degrees C for 1 min. Some plasmid markers are expressed immediately, whereas others require time for phenotypic expression.
已开发出一种方法,可使铜绿假单胞菌PAO菌株以每受体细胞10^(-6) 的频率被质粒和噬菌体DNA转化。该方法在大致轮廓上与为大肠杆菌开发的方法相似。它包括在营养肉汤中将受体细胞培养至每毫升3 - 5×10^8个,用0.1 M氯化镁洗涤细胞,在0.175 M氯化钙中重悬20分钟,暴露于DNA 1小时,然后在42℃热脉冲1分钟。一些质粒标记物立即表达,而其他标记物则需要时间进行表型表达。