Open AccessZinc ions stimulate electrofusion of Hansenula polymorpha protoplasts
Author(s) -
Schnettler Reiner,
Zimmermann Ulrich
Publication year - 1993
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.1574-6968.1993.tb05933.x
Subject(s) - electrofusion , protoplast , zinc , fusion , ion , magnesium , cell fusion , chemistry , mutant , calcium , clone (java method) , biophysics , biology , nuclear chemistry , biochemistry , materials science , metallurgy , cell , dna , linguistics , philosophy , organic chemistry , gene
Electrofusion of protoplasts of two mutant strains of Hansenula polymorpha resulted in high fusion and hybrid yields when the calcium ions present in the conventional fusion medium replaced by zinc ions. The optimal fusion conditions were an alignment field of 0.4 kV cm −1 strength and 2 MHz frequency for 30 s, followed by two consecutive pulses of 12 kV cm −1 strength and 15 μs duration. With 0.05–0.1 mM zinc ions in the fusion medium an average clone number of 10 4 –10 5 clones per 10 8 input cells was reached. The presence of about 0.6 mM magnesium ions in the zinc fusion medium was essential.