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Study of the glucoamylase promoter in Aspergillus niger using green fluorescent protein

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Author: Santerre Henriksen, A.L. · Even, S. · Müller, C. · Punt, P.J. · Hondel, C.A.M.J.J. van den · Nielsen, J.
Type:article
Date:1999
Institution: Centraal Instituut voor Voedingsonderzoek TNO
Source:Microbiology, 3, 145, 729-734
Identifier: 234971
Keywords: Nutrition · Aspergillus niger · Chemostat · Glucoamylase promoter · Green fluorescent protein · Glucan 1,4 alpha glucosidase · Green fluorescent protein · Maltose · Xylose · Aspergillus niger · Continuous culture · Fungus culture · Growth rate · Mycelium · Nonhuman · Priority journal · Aspergillus niger · Genes, Reporter · Glucan 1,4-alpha-Glucosidase · Green Fluorescent Proteins · Luminescent Proteins · Promoter Regions (Genetics) · Aspergillus niger · Fungi

Abstract

An Aspergillus niger strain expressing a red-shifted green fluorescent protein (GFP) in the cytoplasm under the control of the glucoamylase promoter (PglaA) was characterized with respect to its physiology and morphology. Although xylose acted as a repressor carbon source during batch cultivations, PglaA-driven GFP expression by the glucoamylase promoter could be demonstrated in xylose-limited continuous cultures. In these cultivations, the xylose concentration was therefore too low to cause repression. Transient experiments initiated with a maltose pulse did not further induce red-shifted GFP production in xylose-limited continuous cultures. Maltose induction under conditions of xylose repression was microscopically observed and quantified in a flow-through chamber. Red-shifted GFP was first produced after 5 h induction. Finally the strain was characterized in glucose-limited continuous cultures, and here the area of the mycelium stained with cytoplasmic GFP increased with increasing specific growth rate, indicating that GFP can be used as a marker of cellular activity in this type of cultivation.