Figure 1.—

Construction of mak-2 and pp-1 gene replacement mutants. (A) Physical map of the mak-2 genomic region and gene replacement vector pBP-KOMAK2. The XhoI site introduced from primer MAK1 is indicated with an asterisk. PDL37 and PDL38 are primers used to screen Δmak-2 knockout mutants. The indicated 1.5-kb KpnI fragment was used as a hybridization probe for Southern blot analysis. The restriction sites are X, XhoI; K, KpnI; H, HindIII; B, BamHI. (B) Physical map of pp-1 genomic region and gene replacement vector pBP-KOSTE12. STE11, STE14, STEK4, and CTRP2 are primers used to screen Δpp-1 knockout mutants. (C) Southern analysis of Δmak-2 knockout mutant strains PBM5, PBM7, and PBM49. Genomic DNAs were digested with BamHI (B) or KpnI (K) yielding fragments of the indicated sizes (kb). (D) Southern analysis of the Δpp-1 knockout mutant strain DL14. Genomic DNAs were digested with XhoI (X) or KpnI (K) to yield fragments of the indicated sizes.