The sea pansy Renilla reniformis luciferase serves as a sensitive bioluminescent reporter for differential gene expression in Candida albicans.
The infectious yeast Candida albicans progresses through two developmental programs which involve differential gene expression, the bud-hypha transition and high-frequency phenotypic switching. To understand how differentially expressed genes are regulated in this organism, the promoters of phase-specific genes must be functionally characterized, and a bioluminescent reporter system would facilitate such characterization. However, C. albicans has adopted a nontraditional codon strategy that involves a tRNA with a CAG anticodon to decode the codon CUG as serine rather than leucine. Since the luciferase gene of the sea pansy Renilla reinformis contains no CUGs, we have used it to develop a highly sensitive bioluminescent reporter system for C. albicans. When fused to the galactose-inducible promoter of GAL1, luciferase activity is inducible; when fused to the constitutive EF1 alpha 2 promoter, luciferase activity is constitutive; and when fused to the promoter of the white-phase-specific gene WH11 or the opaque-phase-specific gene OP4, luciferase activity is phase specific. The Renilla luciferase system can, therefore, be used as a bioluminescent reporter to analyze the strength and developmental regulation of C. albicans promoters.
Department of Biological Sciences, University of Iowa, Iowa City 52242, USA.
T
T
Srikantha
A
A
Klapach
W W
WW
Lorenz
L K
LK
Tsai
L A
LA
Laughlin
J A
JA
Gorman
D R
DR
Soll
D000818:Animals; D001483:Base Sequence; D002176:Candida albicans / Q000235:genetics* / Q000502:physiology; D003063:Cnidaria / Q000201:enzymology*; D004271:DNA, Fungal / Q000032:analysis; D004274:DNA, Recombinant / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D005690:Galactose / Q000494:pharmacology; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012331:RNA, Fungal / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis
D004271:DNA, Fungal; D004274:DNA, Recombinant; D005656:Fungal Proteins; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D012331:RNA, Fungal; D012333:RNA, Messenger; C083140:Wh11 protein, Candida albicans; D008156:Luciferases; D005690:Galactose