Open AccessLarge‐scale production of yeast hybrids by electrofusion
Author(s) -
Schnettler R.,
Zimmermann U.,
Emeis C.C.
Publication year - 1984
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1984.tb01249.x
Subject(s) - electrofusion , plasmid , saccharomyces cerevisiae , yeast , protoplast , escherichia coli , biology , dielectrophoresis , electroporation , auxotrophy , genetics , microbiology and biotechnology , chemistry , gene , materials science , metallurgy , electrode
Large‐scale production of electrically fused yeast protoplasts from Saccharomyces cerevisiae AH 22[pADH 040‐2] and S. cerevisiae AH215 was achieved by the use of the so‐called helical fusion chamber. Both strains were of the same mating type a and carried the following auxotrophic markers: his4 in the case of AH22 and leu2, his3 in the case of AH215. AH22 also is a carrier of the plasmid pADH 040‐2. This plasmid confers the leu2 gene of yeast and the β‐lactamase gene from Escherichia coli , and this feature enables quick detection of plasmid‐positive cells. After dielectrophoresis (275 V/cm, 800 kHz) fusion was induced by two field pulses (10 kV/cm, 10 μs duration) applied at an interval of 0.5 s. 50 to 60 hybrids per run were isolated after regeneration on selection medium.