Usefulness of heterologous promoters in the Pseudozyma flocculosa gene expression system.
The basidiomycetous fungus Pseudozyma flocculosa represents a promising new host for the expression of complex recombinant proteins. Two novel heterologous promoter sequences, the Ustilago maydis glyceraldehyde-3-phosphate dehydrogenase (GPD) and Pseudozyma tsukubaensis alpha-glucosidase promoters, were tested for their ability to provide expression in P. flocculosa. In liquid medium, these two promoters produced lower levels of intracellular green fluorescent protein (GFP) as compared to the U. maydis hsp70 promoter. However, GPD and alpha-glucosidase sequences behaved as constitutive promoters whereas the hsp70 promoter appeared to be morphology-dependent. When using the hsp70 promoter, the expression of GFP increased proportionally to the concentration of hygromycin in the culture medium, indicating possible induction of the promoter by the antibiotic. Optimal solid-state culture conditions were designed for high throughput screening of hygromycin-resistant transformants with the hsp70 promoter in P. flocculosa.
Avis
Tyler J
TJ
Department of Phytology, Horticulture Research Centre, Envirotron, University Laval, Quebec, Canada, G1K 7P4.
Anguenot
Raphaël
R
Neveu
Bertrand
B
Bolduc
Sébastien
S
Zhao
Yingyi
Y
Cheng
Yali
Y
Labbé
Caroline
C
Belzile
François
F
Bélanger
Richard R
RR
D001483:Base Sequence; D001487:Basidiomycota / Q000235:genetics*; D002934:Cinnamates / Q000494:pharmacology; D017931:DNA Primers; D004352:Drug Resistance, Microbial; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D000520:alpha-Glucosidases / Q000235:genetics*
D002934:Cinnamates; D017931:DNA Primers; D018840:HSP70 Heat-Shock Proteins; D049452:Green Fluorescent Proteins; D006921:Hygromycin B; C026273:hygromycin A; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D000520:alpha-Glucosidases