Isolation and transformation of uracil auxotrophs of the edible basidiomycete Pleurotus ostreatus.
Uracil auxotrophs of Pleurotus ostreatus were isolated using the selectable marker, resistance to 5'-fluoro-orotic acid (5'-FOA). Two of the nine uracil auxotrophs obtained were transformed to prototrophy using plasmid pTRura 3-2 that contains the orotidine monophosphate decarboxylase (ura3) gene from Trichoderma reesei. Southern blot analyses of the transformants showed that the transforming DNA had integrated into the genome of the protoplasts. Using 2 x 10(7) protoplasts, this system gave a transformation efficiency of about 30 transformants per microg of DNA. Normal fruiting bodies were induced in the transformants by crossing them with wild-type monokaryons, and the basidiospores collected from these fruiting bodies showed a biased segregation rate to prototrophy. These results indicate the integrated DNA was stably inherited.
Kim
B G
BG
Division of Applied Microbiology, National Institute of Agricultural Science and Technology, Suwon, South Korea. bgkim@niast.go.kr
Magae
Y
Y
Yoo
Y B
YB
Kwon
S T
ST
D001221:Aspartate Carbamoyltransferase / Q000235:genetics; D015139:Blotting, Southern; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004352:Drug Resistance, Microbial; D005656:Fungal Proteins / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D009097:Multienzyme Complexes / Q000235:genetics; D009154:Mutation / Q000528:radiation effects; D009963:Orotic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D020076:Pleurotus / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins; D014170:Transformation, Genetic; D014242:Trichoderma / Q000235:genetics; D014466:Ultraviolet Rays; D014498:Uracil / Q000378:metabolism*
D004271:DNA, Fungal; D005656:Fungal Proteins; D009097:Multienzyme Complexes; D029701:Saccharomyces cerevisiae Proteins; C110837:URA2 protein, S cerevisiae; D014498:Uracil; D009963:Orotic Acid; C001242:5-fluoroorotic acid; D001221:Aspartate Carbamoyltransferase; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)