Toyama H, Anthony C, Lidstrom M E
Department of Biological Chemistry, Yamaguchi University, Japan.
FEMS Microbiol Lett. 1998 Sep 1;166(1):1-7. doi: 10.1111/j.1574-6968.1998.tb13175.x.
Methylobacterium extorquens AM1 is a pink-pigmented facultative methylotroph which is widely used for analyzing pathways of C1 metabolism with biochemical and molecular biological techniques. To facilitate this approach, we have applied a new method to construct insertion or disruption mutants with drug resistance genes by electroporation. By using this method, mutants were obtained in four genes present in the mxa methylotrophy gene cluster for which the functions were unknown, mxaR, mxaS, mxaC and mxaD. These mutants were unable to grow on methanol except the mutant of mxaD, which showed reduced growth on methanol.
嗜甲基甲基杆菌AM1是一种粉红色的兼性甲基营养菌，广泛用于通过生化和分子生物学技术分析C1代谢途径。为了便于采用这种方法，我们应用了一种新方法，通过电穿孔构建带有耐药基因的插入或破坏突变体。利用这种方法，在mxa甲基营养基因簇中存在的四个功能未知的基因mxaR、mxaS、mxaC和mxaD中获得了突变体。除了mxaD突变体在甲醇上生长有所减少外，这些突变体在甲醇上均无法生长。
