Open AccessIsolation and transformation of uracil auxotrophs of the edible basidiomycete Pleurotus ostreatus
Author(s) -
Kim BeomGi,
Magae Yumi,
Yoo YoungBok,
Kwon SukTae
Publication year - 1999
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.1574-6968.1999.tb08848.x
Subject(s) - pleurotus ostreatus , isolation (microbiology) , uracil , auxotrophy , biology , microbiology and biotechnology , transformation (genetics) , chemistry , food science , biochemistry , mushroom , gene , dna , escherichia coli
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 2×10 7 protoplasts, this system gave a transformation efficiency of about 30 transformants per μg 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.