FIG. 1.

Isolation of an M. pneumoniae prkC transposon insertion mutant. (A) Schematic representation of the genomic region surrounding the prkC gene in M. pneumoniae and site of the transposon insertion in the prkC knockout strain GPM11. The annealing sites of oligonucleotides used for the determination of the transposon insertion site are indicated by arrows. Probes hybridizing to internal fragments of the prkC and the aac-ahpD genes are depicted as dotted lines. (B) Southern blot analysis to confirm the single insertion of the minitransposon into the prkC gene of strain GPM11. Chromosomal DNA of the wild type and of strain GPM11 was digested using XhoI. Blots were hybridized with the prkC-specific probe (left) and a probe hybridizing to the aac-ahpD genes of the minitransposon (right). DIG-labeled DNA molecular size marker III (Roche Applied Science) served as a standard.