Regulated expression of green fluorescent protein in Debaryomyces hansenii.
The broad range of environmental conditions under which Debaryomyces hansenii can grow, and its production of lipolytic and proteolytic enzymes, have promoted its widespread use. The present work represents a preliminary characterization of D. hansenii for heterologous expression and secretion of green fluorescent protein (GFP). Six heterologous expression vectors were used to address protein production efficiency under regulated expression conditions. Protein expression in D. hansenii seems to be similar to that in Saccharomyces cerevisiae, with transcription being controlled by almost all of the S. cerevisiae and D. hansenii inducible promoters tested, with the exception of the alcohol dehydrogenase 2 gene promoter from S. cerevisiae. Extracellular protein levels in D. hansenii were lower than in S. cerevisiae when Saccharomyces signal peptides were used.
Maggi
Ricardo G
RG
Department of Marine Sciences, Marine Station, University of Puerto Rico, P.O. Box 908, Lajas, PR 00667, USA. rgmaggi@ncsu.edu
Govind
Nadathur S
NS
D000426:Alcohol Dehydrogenase / Q000235:genetics; D045304:Cytochromes c / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D005822:Genetic Vectors; D005993:Glycerolphosphate Dehydrogenase / Q000235:genetics; D049452:Green Fluorescent Proteins; D006360:Heat-Shock Proteins / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011494:Protein Kinases / Q000235:genetics; D021382:Protein Sorting Signals; D004718:Saccharomycetales / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic
D006360:Heat-Shock Proteins; D008164:Luminescent Proteins; D021382:Protein Sorting Signals; D049452:Green Fluorescent Proteins; D045304:Cytochromes c; D005993:Glycerolphosphate Dehydrogenase; D000426:Alcohol Dehydrogenase; D011494:Protein Kinases