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. 2005 Jun;4(6):999–1008. doi: 10.1128/EC.4.6.999-1008.2005

FIG. 1.

FIG. 1.

Confirmation analyses of RIM101 mutation. A. DNA was subjected to PCR with primers 2075 and 2078. Lanes 1 to 3, Δrim101 mutants (BMA1, BMA2, and BMA4, respectively); lane 4, FB2 wild-type strain; lane 5, plasmid pUpacH; lane 6, plasmid pUpacC4. B. EcoRI digestion of PCR products. Lane 2, BMA1; lane 4, BMA2; lane 6, FB2; lane 8, pUpacH; lane 10, pUpacC4. Lanes 1, 3, 5, 7, and 9, controls with no restriction enzyme added to BMA1, BMA2, FB2, pUpacH, and pUpacC4, respectively. C. Southern blot hybridization of HincII-digested DNA with the 1.2-kb StuI-BamHI RIM101-specific probe. Lane 1, FB1; lane 2, BMA1; lane 3, BMA7; lane 4, FB2; lane 5, BMA2; lane 6, BMA4. Note absence of hybridization in mutants compared to wild-type strains. Arrows show positions of the molecular size standards. D. Restriction map of RIM101 locus of the wild-type strain showing the primer (solid arrow) and probe (solid line) positions. E. Restriction map of RIM101 locus of the mutant allele used in the experiment showing the position of the primers (solid arrows) and the Hph gene (large open arrow).