Volume 13
Issue 2
Biotechnology
JOURNAL OF
POLISH
AGRICULTURAL
UNIVERSITIES
Available Online: http://www.ejpau.media.pl/volume13/issue2/abs-08.html
YARROWIA LIPOLYTICA CELL CYCLE STUDY
Agnieszka Mituła1, Wojciech Barszczewski2, Małgorzata Robak2
1 Institute of Biochemistry and Molecular Biology,
University of Wrocław, Poland
2 Department of Biotechnology and Food Microbiology,
Wrocław University of Environmental and Life Sciences, Poland
ABSTRACT
Flow cytometry (FC) was used for measuring ploidity and number of cells at each phase of the cell cycle of four Yarrowia lipolytica yeast strains. Synchronization at G0/G1-phase was partially achieved by carbon source starvation, followed by glucose addition. Approximately 10 000 cells were analyzed by measuring the fluorescence of propidium iodine (PI) bound to DNA after 0, 1, 2, 4, 6 hours of glucose addition to the starved cells population. Estimation of cells percentage at each cycle phase was achieved by Cylchred and WinMDI analysis of obtained data. However complete synchronization was not obtained for any of the strains, the generation time representing time interval between
two maxima of cells at S phase was determined. The shortest generation time was noted for Y. lipolytica AWG7 (1h). Comparison of histograms of all tested strains with a haploid type strain roved their haploidy.
Key words: flow cytometry, cell cycle, yeasts, Yarrowia lipolytica.
Agnieszka Mituła
Institute of Biochemistry and Molecular Biology,
University of Wrocław, Poland
Wojciech Barszczewski
Department of Biotechnology and Food Microbiology,
Wrocław University of Environmental and Life Sciences, Poland
Norwida 25, 50-373 Wrocław, Poland
Phone: +48 71 3205463
email: wojciech.barszczewski@up.wroc.pl
Małgorzata Robak
Department of Biotechnology and Food Microbiology,
Wrocław University of Environmental and Life Sciences, Poland
Chełmońskiego 37/41
51-630 Wrocław
Poland
email: malgorzata.robak@up.wroc.pl
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