Highly-efficient transformation of the homobasidiomycete Schizophyllum commune to phleomycin resistance.
Regulatory sequences of the glyceraldehyde-3-phosphate-dehydrogenase (GPD) gene from the homobasidiomycete Schizophyllum commune were fused to the coding sequence of the ble gene from Streptoalloteichus hindustanus, which codes for a phleomycin-binding protein. The resulting construct transformed S. commune to phleomycin resistance at a high frequency (up to 10(4) transformants/microgram DNA per 10(7) protoplasts) when regeneration was done in 0.5 M MgSO4. A similar construct with regulatory sequences from Aspergillus nidulans failed to give transformants, showing the importance of homologous regulatory sequences for the expression of genes in S. commune. The homologous GPD promoter could be deleted up to position -130 without any effect on the number of phleomycin-resistant transformants. This is the first effective stable transformation system in a homobasidiomycete employing antibiotic resistance.
Schuren
F H
FH
Department of Plant Biology, University of Groningen, Haren, The Netherlands.
Wessels
J G
JG
D000192:Actinomycetales / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics*; D015870:Gene Expression; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis / Q000235:genetics*; D010692:Phleomycins / Q000494:pharmacology; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011523:Protoplasts / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D012567:Schizophyllum / Q000187:drug effects / Q000235:genetics*; D014158:Transcription, Genetic; D014170:Transformation, Genetic
D010692:Phleomycins; D011993:Recombinant Fusion Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases