Moore A T, Nayudu M, Holloway B W
J Gen Microbiol. 1983 Mar;129(3):785-99. doi: 10.1099/00221287-129-3-785.
Prime plasmids have been isolated from Methylophilus methylotrophus AS1 using the IncP-1 plasmid pMO172. Such plasmids can complement mutant function when transferred to appropriate strains of Pseudomonas aeruginosa PAO. From the range of particular functions complemented by each prime plasmid, a preliminary map of the M. methylotrophus AS1 genome with four groups of linked markers was obtained. Physical examination of two of these prime plasmids showed that each had acquired an additional DNA segment. Southern hybridization experiments showed there was sequence homology between the additional DNA segments and M. methylotrophus AS1 DNA, confirming that these prime plasmids carried a segment of the M. methylotrophus AS1 genome. Mapping by complementation may be applicable to other bacteria in which mutants suitable for selecting recombinants are not readily available.
已使用IncP-1质粒pMO172从甲基营养型嗜甲基菌AS1中分离出起始质粒。当将这些质粒转移到铜绿假单胞菌PAO的合适菌株中时,它们可以补充突变体的功能。根据每个起始质粒所补充的特定功能范围,获得了带有四组连锁标记的嗜甲基菌AS1基因组的初步图谱。对其中两个起始质粒的物理检查表明,每个质粒都获得了一个额外的DNA片段。Southern杂交实验表明,额外的DNA片段与嗜甲基菌AS1 DNA之间存在序列同源性,证实这些起始质粒携带了嗜甲基菌AS1基因组的一个片段。通过互补进行定位可能适用于其他难以获得适合选择重组体的突变体的细菌。