tailieunhanh - Electrofusion of Saccharomyces cerevisiae auxotrophic mutants of identical mating type using a laboratory made system

Yeast protoplasts from haploid auxotroph strains DC6 (MATα) and FY73 (MATα) were exposed to an inhomogeneous alternating field (AC 200 V/cm). Due to the dielectrophoretic aggregation two or more cells with close membrane contact were formed between the two electrodes. | Turk J Biol 27 (2003) 1-5 © TÜB‹TAK Electrofusion of Saccharomyces cerevisiae Auxotrophic Mutants of Identical Mating Type Using a Laboratory-Made System Gökhan CORAL* Department of Biology, Faculty of Arts and Sciences, Mersin University, 33342 Çiftliköy-Mersin - TURKEY Ömer ÇOLAK Department of Biology, Faculty of Arts and Sciences, Çukurova University, 01330 Balcal›-Adana - TURKEY M. Nisa ÜNALDI Department of Biology, Faculty of Arts and Sciences, Mustafa Kemal University, 31040 Hatay - TURKEY Burhan ARIKAN Department of Biology, Faculty of Arts and Sciences, Çukurova University, 01330 Balcal›-Adana - TURKEY Received: Abstract: Yeast protoplasts from haploid auxotroph strains DC6 (MATα) and FY73 (MATα) were exposed to an inhomogeneous alternating field (AC 200 V/cm). Due to the dielectrophoretic aggregation two or more cells with close membrane contact were formed between the two electrodes. The effects of different DC pulses were investigated to determine critical fusogenic pulse strength. In the results, the maximum fusant cells were obtained only when 8 kV field strengths were applied. These findings showed that the optimum fusogenic pulse strength for yeast protoplasts is 8 kV/cm under our experimental conditions. After fusion, the DNA contents of both hybrid and individual parental strains were compared. The amount of DNA in the hybrid was found to be about twofold higher than that in the individual parental strain. Key Words: Electrofusion, Saccharomyces cerevisiae ‹dentik Eflleflme Tipine Sahip Saccharomyces cerevisiae Oksotrofik Mutantlar›n›n Laboratuvar Yap›m› Bir Sistem Kullan›larak Elektrofüzyonu Özet: Bu çal›flmada DC6 (MATα) ve FY73 (MATα) haploid oksotrof maya sufllar›ndan elde edilen protoplastlar, homojen olmayan bir alternatif alana (200 V/cm AC) maruz b›rak›ld›. Dielektroforetik agregasyona ba¤l› olarak, iki elektrot aras›nda kalan iki ya da daha fazla say›daki hücrede yak›n membran temas› oluflturuldu. Hücreleri füzyona .