Usefulness of heterologous promoters in the Pseudozyma flocculosa gene expression system

• Published in: Bioscience, biotechnology, and biochemistry Vol. 72; no. 2; pp. 456 - 462
• Main Authors: Avis, T.J.(Laval Univ., Quebec (Canada)), Anguenot, R, Neveu, B, Bolduc, S, Zhao, Y, Cheng, Y, Labbe, C, Belzile, F, Belanger, R.R
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.02.2008; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: alpha-Glucosidases - genetics; Base Sequence; Basidiomycota - genetics; BASIDIOMYCOTINA; Biological and medical sciences; Cinnamates - pharmacology; constitutive promoter; DNA Primers; Drug Resistance, Microbial; EXPRESION GENICA; EXPRESSION DES GENES; Fundamental and applied biological sciences. Psychology; GENE EXPRESSION; gene expression system; GENETICALLY MODIFIED MICROORGANISMS; GLICOSIDASAS; Glyceraldehyde-3-Phosphate Dehydrogenases - genetics; GLYCOSIDASE; GLYCOSIDASES; Green Fluorescent Proteins - genetics; heterologous promoter; HSP70 Heat-Shock Proteins - genetics; Hygromycin B - analogs & derivatives; Hygromycin B - pharmacology; inducible promoter; MICRO-ORGANISME MODIFIE GENETIQUE; MICROORGANISMOS MODIFICADOS GENETIC; NUCLEOTIDE SEQUENCE; Promoter Regions, Genetic; PROTEINAS RECOMBINANTES; PROTEINE RECOMBINANTE; Pseudozyma; Pseudozyma flocculosa; RECOMBINANT PROTEINS; SECUENCIA NUCLEOTIDICA; SEQUENCE NUCLEOTIDIQUE; Ustilago maydis; USTILAGO ZEAE; Pseudozyma flocculosa; heterologous promoter; constitutive promoter; gene expression system; Gene expression; inducible promoter
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.70563

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.