« Previous AbstractFlexible origin of hydrocarbon/pheromone precursors in Drosophila melanogaster    Next AbstractDimorphism and haploid fruiting in Cryptococcus neoformans: association with the alpha-mating type »

Mol Microbiol

Title:		The Cryptococcus neoformans GAL7 gene and its use as an inducible promoter
Author(s):		Wickes BL; Edman JC;
Address:		"Department of Laboratory Medicine, University of California, San Francisco 94143-0626, USA"
Journal Title:		Mol Microbiol
Year:		1995
Volume:		16
Issue:		6
Page Number:		1099 - 1109
DOI:		10.1111/j.1365-2958.1995.tb02335.x
ISSN/ISBN:		0950-382X (Print) 0950-382X (Linking)
Abstract:		"A Cryptococcus neoformans galactose auxotroph was created by ultraviolet light mutagenesis and complemented with a C. neoformans genomic library. The translated sequence of the complementing DNA revealed a high degree of similarity to a number of UDP glucose-D-galactose-1-phosphate uridylyltransferases. Expression of C. neoformans GAL7 mRNA followed a pattern similar to Saccharomyces cerevisiae expression; it was first observed within 2.5 min of induction and fully induced by 30 min. The gene was completely repressed in the presence of glucose. The GAL7 promoter was isolated and used to construct a promoter cassette. Two genes were tested in this cassette for galactose regulation by creating GAL7 promoter fusions with their coding regions. MF alpha, which encodes a pheromone, was found to produce filaments only in transformants that were induced by galactose. A second gene, beta-glucuronidase (gusA), which is a commonly used reporter gene, was tested and also found to be expressed. When the GAL7p::GUS fusion was used to quantify inducibility of the GAL7 promoter, the level of enzyme activity was at least 500-fold greater for cells grown in galactose than for cells grown in glucose. The GAL7 promoter is the first inducible promoter characterized in C. neoformans and the GUS gene is the first heterologous gene shown to be expressed in this yeast pathogen"
Keywords:		"Amino Acid Sequence Base Sequence Blotting, Northern Cloning, Molecular Cryptococcus neoformans/*genetics/metabolism DNA Primers Galactose/metabolism Gene Expression Regulation, Fungal/*genetics Glucose/pharmacology Glucose-6-Phosphate Glucosephosphates/b;"
Notes:		"MedlineWickes, B L Edman, J C eng R01 AI29312/AI/NIAID NIH HHS/ Research Support, U.S. Gov't, P.H.S. England 1995/06/01 Mol Microbiol. 1995 Jun; 16(6):1099-109. doi: 10.1111/j.1365-2958.1995.tb02335.x"