Expression of the Klebsiella pneumoniae pullulanase-encoding gene in Saccharomyces cerevisiae

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dc.contributor.authorJanse B.J.H.
dc.contributor.authorPretorius I.S.
dc.date.accessioned2011-05-15T16:01:30Z
dc.date.available2011-05-15T16:01:30Z
dc.date.issued1993
dc.description.abstractA 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 α-factor (MFα1(p)) and alcohol dehydrogenase (ADC1(p)) gene promoters. Secretion of pullulanase was directed by the leader sequence of the yeast mating pheromone α-factor (MFα1(s)). Transcription termination was effected by the yeast tryptophan synthase gene terminator (TRP5(r)). 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.
dc.description.versionArticle
dc.identifier.citationCurrent Genetics
dc.identifier.citation24
dc.identifier.citation02-Jan
dc.identifier.issn1728083
dc.identifier.urihttp://hdl.handle.net/10019.1/12009
dc.subjectmessenger rna
dc.subjectpullulanase
dc.subjectarticle
dc.subjectcloning vector
dc.subjectenzyme assay
dc.subjectenzyme release
dc.subjectgene expression
dc.subjectgene insertion
dc.subjectklebsiella pneumoniae
dc.subjectmolecular cloning
dc.subjectnonhuman
dc.subjectnorthern blotting
dc.subjectplasmid
dc.subjectpriority journal
dc.subjectpromoter region
dc.subjectsaccharomyces cerevisiae
dc.subjectsouthern blotting
dc.subjecttranscription initiation
dc.subjecttranscription termination
dc.subjectBase Sequence
dc.subjectBlotting, Northern
dc.subjectBlotting, Southern
dc.subjectCloning, Molecular
dc.subjectDNA, Fungal
dc.subjectElectrophoresis, Polyacrylamide Gel
dc.subjectGenes, Fungal
dc.subjectGlycoside Hydrolases
dc.subjectKlebsiella pneumoniae
dc.subjectMolecular Sequence Data
dc.subjectPeptides
dc.subjectPolymerase Chain Reaction
dc.subjectRestriction Mapping
dc.subjectSaccharomyces cerevisiae
dc.subjectTerminator Regions (Genetics)
dc.subjectTranscription, Genetic
dc.subjectTransformation, Genetic
dc.subjectTryptophan Synthase
dc.titleExpression of the Klebsiella pneumoniae pullulanase-encoding gene in Saccharomyces cerevisiae
dc.typeArticle
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