Integration of heterologous genes in several yeast species using vectors containing a Hansenula polymorpha-derived rDNA-targeting element.
A method that has been successfully used to generate recombinant Hansenula polymorpha strains by transformation with rDNA-targeting vectors was applied in the present study to a range of alternative yeast hosts, using vectors with an H. polymorpha-derived integration sequence. The dimorphic yeast Arxula adeninivorans, which is currently being assessed for heterologous gene expression, was the main focus of the study. As in H. polymorpha, it was possible to co-integrate more than a single plasmid carrying an expressible gene. Additionally, the vectors were examined in two further species, Pichia stipitis and Saccharomyces cerevisiae. Based on these results the design of a 'universal' fungal vector appears to be feasible.
Klabunde
Jens
J
Institut für Mikrobiologie, Heinrich-Heine-Universität Düsseldorf, Universitätsstr. 1, 40225, Düsseldorf, Germany. klabundj@uni-duesseldorf.de
Kunze
Gotthard
G
Gellissen
Gerd
G
Hollenberg
Cornelis P
CP
D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004271:DNA, Fungal; D004274:DNA, Recombinant / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal / Q000235:genetics; D017930:Genes, Reporter; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D016681:Genome, Fungal; D016366:Open Reading Frames; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D004718:Saccharomycetales / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D014170:Transformation, Genetic
D004271:DNA, Fungal; D004274:DNA, Recombinant; D004275:DNA, Ribosomal; D005656:Fungal Proteins; D011994:Recombinant Proteins