Electrofusion of Trichoderma reesei protoplasts |
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Authors: | M MURKOVIC W STEINER H ESTERBAUER |
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Institution: | Institute of Biotechnology, Microbiology and Waste Treatment, Technical University of Graz, Schlögelgasse 9,8010 Graz;*Institute of Biochemistry, University of Graz, Schubertstrasse 1, 8010 Graz, Austria |
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Abstract: | Protoplasts of Trichoderma reesei were fused according to the method of Zimmermann. For optimizing the fusion parameters the central composite design was used. Genetic evidence for fusion has been obtained by segregation of the auxotrophic markers in the haploid conidia. The parameters which were optimized were: pulse voltage, pulse duration and number of pulses. The optimal parameters for the fusion of T. reesei protoplasts are 90 V pulse voltage, 37 μS pulse duration and six pulses at intervals of 1-0 s. |
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