Induction of mating in Candida albicans by construction of MTLa and MTLα strains

Science

Volumn 289, Issue 5477, 2000, Pages 310-313

  Magee, B B ^a   Magee, P T ^a  

^a UNIVERSITY OF MINNESOTA   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ALLELE; ARTICLE; CANDIDA ALBICANS; DIPLOIDY; DNA CONTENT; GENE LOCUS; MATING; NONHUMAN; PRIORITY JOURNAL; REPRODUCTION; STRAIN DIFFERENCE; TETRAPLOIDY;

CANDIDA ALBICANS; CHROMOSOMES, FUNGAL; DNA, FUNGAL; GENES, FUNGAL; GENES, MATING TYPE, FUNGAL; PLOIDIES; RECOMBINATION, GENETIC; SACCHAROMYCES CEREVISIAE; SORBOSE; SPECIES SPECIFICITY;

CANDIDA ALBICANS; FUNGI; SACCHAROMYCES;

EID: 0034647921     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.289.5477.310     Document Type: Article

References (32)

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21. 7/ml) were fixed in 70% alcohol, washed with phosphate-buffered saline (PBS), treated with ribonuclease (1 mg/ml, 37°C, 75 min), and washed again with PBS. Cells were stained with propidium iodide (50 μg/ml) shortly before sorting (24, 25). FACS analysis was done on an Ortho Cytofluorograf lls (Diagnostic Systems) operated at 488 nm and 100 mW laser power. Propidium iodide fluorescence was collected with a 570 long-pass filter. Data were collected in list, area, and linear modes at rates of a few hundred cells per second with the Cicero Data acquisition system (Cytomation, Fort Collins, CO) (26).
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31. The pulsed-field gel was run under conditions that emphasize the smaller chromosomes: 0.9% agarose, 0.5x tris-borate EDTA, 60-to 120-s switch, 6 V/cm, 120°C, 24 hours, 15°C, run on a CHEF DRIII (Biorad). Cells were grown on YEPD. Chromosomes were prepared as described (27).
32. N. Abu-Absi and F. Srienc helped with the FACS analysis. S. Scherer, J. Beckerman, H. Chibana, and S. Grindle contributed helpful criticism, and J. Berman and S. Scherer provided comments on the manuscript C. Hull and A. Johnson generously provided the sequence of the MTLa allele and the organization of the MTL region before publication. We are very grateful to D. Gartner and the College of Biological Sciences Imaging Center (University of Minnesota) for help with the figures. Supported by grants AI16567, AI35109, and AI46351 from NIH (P.T.M.).