Expression plasmids for use in Candida glabrata.
We describe a series of CEN/ARS episomal plasmids containing different Candida glabrata promoters, allowing for a range of constitutive or regulated expression of proteins in C. glabrata. The set of promoters includes three constitutive promoters (EGD2pr, HHT2pr, PDC1pr), two macrophage/phagocytosis-induced promoters (ACO2pr, LYS21pr), and one nutritionally regulated promoter (MET3pr). Each promoter was cloned into two plasmid backbones that differ in their selectable marker, URA3, or the dominant-selectable NAT1 gene, which confers resistance to the drug nourseothricin. Expression from the 12 resulting plasmids was assessed using GFP as a reporter and flow cytometry or quantitative reverse-transcription polymerase chain reaction to assess expression levels. Together this set of plasmids expands the toolkit of expression vectors available for use with C. glabrata.
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Rebecca E
RE
Zordan
Yuxia
Y
Ren
Shih-Jung
SJ
Pan
Giuseppe
G
Rotondo
Alejandro
A
De Las Peñas
Joseph
J
Iluore
Brendan P
BP
Cormack
D041221:Candida glabrata / Q000235:genetics* / Q000378:metabolism; D004274:DNA, Recombinant / Q000235:genetics* / Q000378:metabolism; D017930:Genes, Reporter; D005819:Genetic Markers; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic
D004274:DNA, Recombinant; D005819:Genetic Markers
Candida glabrata; MET3; expression vector; inducible; macrophage