Electrofusion of Saccharomyces cerevisiae Auxotrophic Mutants of Identical Mating Type Using a Laboratory-Made System

Coral, Mutlu Nisa Ünaldı | Coral, Gökhan

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...