Isolation of uracil auxotrophs of the fungus Acremonium cellulolyticus and the development of a transformation system with the pyrF gene.
Acremonium cellulolyticus CF-2612 is a cellulase hyper-producing mutant that originated from A. cellulolyticus Y-94. In this study, we isolated a uracil auxotroph (strain CFP3) derived from CF-2612, and cloned a wild-type pyrF gene encoding orotate phosphoribosyl transferase (OPRTase) from Y-94. OPRTase activity was not detected in strain CFP3, which had one nucleotide substitution in its pyrF gene. The wild-type pyrF gene restored the defective growth of CFP3 on uracil-free medium, and PCR and Southern analyses revealed that wild-type pyrF was integrated into the genome. These results indicate that our transformation system for A. cellulolyticus with the pyrFgene as a selection marker was successful.
Biomass Technology Research Center, National Institute of Advanced Industrial Science and Technology, Hiroshima, Japan. tatsuya.fujii@aist.go.jp
Tatsuya
T
Fujii
Kazuya
K
Iwata
Katsuji
K
Murakami
Shinichi
S
Yano
Shigeki
S
Sawayama
D000164:Acremonium / Q000235:genetics / Q000378:metabolism*; D005800:Genes, Fungal; D014170:Transformation, Genetic; D014498:Uracil / Q000378:metabolism*
D014498:Uracil