Fig 4.

Genotyping of the pyrE, aldC, and als loci of KGMA0119 (wild type) and its derivatives. (A to C, upper blots) Southern blot analyses. Genomic DNAs of the four strains were digested with PuvII (A) or SmaI (B and C) and hybridized with the pyrE probe (A) or the aldC probe (B and C). (A to C, lower blots) PCR analyses. The pyrE (A), aldC (B), and als (C) loci of the four strains were amplified from genomic DNA using primer pairs EP-F3–E-R4 (A), aldC-F2–aldC-R2 (B), and als-F2–ldh-i2 (C). In the case of als disruption, the PCR products were digested with EcoRI to confirm the targeted gene disruption. Figure 3 shows the restriction map and the position of each primer.