Cloning and sequencing of the ura3 locus of the methylotrophic yeast Hansenula polymorpha and its use for the generation of a deletion by gene replacement.
The ura3 gene of Hansenula polymorpha was cloned, sequenced and used to generate a ura3 mutant from the wild-type strain of this yeast via integrative mutagenesis. The Tn5 neomycin-resistance marker (neo) under control of the ADH1 promoter from Saccharomyces cerevisiae served as a transformation marker. The results show that gene replacement can be achieved in H. polymorpha, a yeast with a high level of non-homologous integration.
Merckelbach
A
A
Rhein Biotech GmbH, Düsseldorf, Germany.
Gödecke
S
S
Janowicz
Z A
ZA
Hollenberg
C P
CP
D003001:Cloning, Molecular; D017353:Gene Deletion; D005800:Genes, Fungal / Q000235:genetics*; D005819:Genetic Markers; D016297:Mutagenesis, Site-Directed; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000235:genetics*; D010843:Pichia / Q000201:enzymology / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic / Q000235:genetics*
D005819:Genetic Markers; D009964:Orotidine-5'-Phosphate Decarboxylase