Transformation system for a wastewater treatment yeast, Hansenula fabianii J640: isolation of the orotidine-5'-phosphate decarboxylase gene (URA3) and uracil auxotrophic mutants.
A transformation system for Hansenula fabianii J640, a commonly used wastewater treatment yeast, was constructed. As a host cell, a uracil auxotrophic mutant designated as H. fabianii J640 u-1, which was confirmed to have a mutation at the locus of the gene for orotidine-5'-phosphate (OMP) decarboxylase (URA3), was obtained by positive selection using 5-fluoroorotic acid. A plasmid named pHFura3, which includes a 795-bp open-reading frame of the OMP decarboxylase H. fabianii, was obtained by complementation of the Escherichia coli pyrF mutant, pHFura3 could transform H. fabianii J640 u-1 by a non-homologous and frequently multicopy integration into the host genomic DNA.
Kato
M
M
National Research Institute of Brewing, Higashi-Hiroshima, Japan.
Iefuji
H
H
Miyake
K
K
Iimura
Y
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D016681:Genome, Fungal; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000235:genetics*; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic; D014498:Uracil / Q000378:metabolism*
D004271:DNA, Fungal; D014498:Uracil; D009964:Orotidine-5'-Phosphate Decarboxylase