Transformation system for a wastewater treatment yeast, Hansenula fabianii J640: isolation of the orotidine-5'-phosphate decarboxylase gene (URA3) and uracil auxotrophic mutants

Applied Microbiology and Biotechnology
M KatoY Iimura

Abstract

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.

Citations

Jul 8, 2008·Applied Microbiology and Biotechnology·Takashi WatanabeHaruyuki Iefuji
Oct 18, 2006·FEMS Yeast Research·Kousuke KurodaYoshifumi Jigami
Sep 13, 2011·AMB Express·Miyoshi Kato, Haruyuki Iefuji
May 1, 2007·Journal of Microbiological Methods·Yuriy R BoretskyAndriy A Sibirny
Oct 16, 2001·Critical Reviews in Biotechnology·T T WangB H Lee

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