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Curr Genet

Title:		Expression of the Klebsiella pneumoniae pullulanase-encoding gene in Saccharomyces cerevisiae
Author(s):		Janse BJ; Pretorius IS;
Address:		"Department of Microbiology, University of Stellenbosch, South Africa"
Journal Title:		Curr Genet
Year:		1993
Volume:		24
Issue:		1-Feb
Page Number:		32 - 37
DOI:		10.1007/BF00324662
ISSN/ISBN:		0172-8083 (Print) 0172-8083 (Linking)
Abstract:		A 3800-base pair (bp) DNA fragment encoding the mature pullulanase from Klebsiella pneumoniae was inserted between two different yeast expression-secretion cassettes and an yeast gene terminator. These cassettes were cloned into an yeast centromeric plasmid YCplacIII and transformed into laboratory strains of Saccharomyces cerevisiae. Transcription initiation signals were derived from the mating pheromone alpha-factor (MF alpha 1p) and alcohol dehydrogenase (ADC1p) gene promoters. Secretion of pullulanase was directed by the leader sequence of the yeast mating pheromone alpha-factor (MF alpha 1s). Transcription termination was effected by the yeast tryptophan synthase gene terminator (TRP5T). Southernblot analysis confirmed the presence of pulA in transformed yeasts and Northern-blot analysis revealed the presence of PUL1 mRNA. A pullulan agarose assay indicated the extracellular production of biologically active pullulanase by S. cerevisiae
Keywords:		"Base Sequence Blotting, Northern Blotting, Southern Cloning, Molecular DNA, Fungal Electrophoresis, Polyacrylamide Gel *Genes, Fungal Glycoside Hydrolases/biosynthesis/*genetics Klebsiella pneumoniae/enzymology/*genetics Mating Factor Molecular Sequence D;"
Notes:		"MedlineJanse, B J Pretorius, I S eng 1993/07/01 Curr Genet. 1993 Jul-Aug; 24(1-2):32-7. doi: 10.1007/BF00324662"