Chen Ding-Qiang, Huang Shao-Song, Lu Yong-Jun
Department of Biochemistry and State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen Zhongshan University, Guangzhou, China.
Microbiol Res. 2006;161(3):246-51. doi: 10.1016/j.micres.2005.09.001. Epub 2005 Nov 11.
A simple and reproducible method has been developed to transform Legionella pneumophila by electroporation. Effects of different conditions, including electric field strength, pulse length, DNA quality and cell density, were evaluated. Using our method, an efficiency of up to 6 x 10(7) transformants/microg DNA was obtained. This optimized transformation procedure should efficiently facilitate gene manipulations in L. pneumophila, such as plasmid transfer, transposon mutagenesis, library transformation for complementation cloning, etc.
已经开发出一种通过电穿孔转化嗜肺军团菌的简单且可重复的方法。评估了不同条件的影响,包括电场强度、脉冲长度、DNA质量和细胞密度。使用我们的方法,获得了高达6×10⁷转化子/μg DNA的效率。这种优化的转化程序应能有效地促进嗜肺军团菌中的基因操作,如质粒转移、转座子诱变、用于互补克隆的文库转化等。