Candida glabrata shuttle vectors suitable for translational fusions to lacZ and use of beta-galactosidase as a reporter of gene expression.
The functionality of beta-galactosidase encoded by the E. coli lacZ gene as a reporter of gene expression in C. glabrata was investigated. C. glabrata/E. coli shuttle vectors were constructed, containing both a C. glabrata CEN-ARS cassette, to allow regular segregation and episomal replication of the plasmids, and the lacZ coding sequence of E. coli. The functionality of beta-galactosidase in C. glabrata was verified by inserting the promoter and the 5' coding region of the HIS3 gene from C. glabrata directionally upstream of the lacZ gene. By fusing the promoter of the copper-controlled MTII gene to the lacZ reporter, we showed that beta-galactosidase activity can be differentially induced in C. glabrata. beta-galactosidase reporter activities were detected qualitatively by an indirect filter assay and quantitatively from permeabilized cells.
El Barkani
A
A
Institut für Hygiene und Mikrobiologie, Universität Würzburg, Josef-Schneider-Strasse 2, 97080, Würzburg, Germany.
Haynes
K
K
Mösch
H
H
Frosch
M
M
Mühlschlegel
F A
FA
D002175:Candida / Q000235:genetics*; D002503:Centromere / Q000235:genetics; D019327:Copper Sulfate / Q000494:pharmacology; D004271:DNA, Fungal / Q000235:genetics; D004274:DNA, Recombinant; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006836:Hydro-Lyases / Q000235:genetics; D007763:Lac Operon / Q000235:genetics*; D008668:Metallothionein / Q000235:genetics; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D012093:Replicon / Q000235:genetics; D001616:beta-Galactosidase / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism
D004271:DNA, Fungal; D004274:DNA, Recombinant; D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D008668:Metallothionein; D001616:beta-Galactosidase; D006836:Hydro-Lyases; C021434:imidazoleglycerolphosphate dehydratase; D019327:Copper Sulfate