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View full-text article in PMC

. 2009 Jul 27;4(7):e6321. doi: 10.1371/journal.pone.0006321

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Willger et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Serotype D strains pmt1D (MATa pmt1D::URA5 ade2-27) and pmt4D (MATα pmt4D::ADE2 ura5) were crossed on standard V8 mating media and individual spores were isolated after several days by micromanipulation. A: 15 individual progeny were analyzed by colony PCR for the presence of the PMT1 (upper panel) and PMT4 alleles (lower panel). Position of the respective wild-type and disruption alleles are indicated at right. Wild-type strain JEC21 was used as a control. B: The 15 strains from A were spotted onto the indicated plates, grown for 2–3 days at 30°C and subsequently analyzed for auxotrophic marker distribution. Strains were spotted from top left (#1) to bottom right (wild-type control). C: Summary of the genotypes identified in A and B with respect to PMT alleles and mating-type. Mating-type of the individual spores were determined by standard mating reactions using wild-type strains JEC20 (MATa) and JEC21 (MATα) as tester strains.