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PMID: 9784536 Published · ppublish English Journal Article

Altered expression of selectable marker URA3 in gene-disrupted Candida albicans strains complicates interpretation of virulence studies.

Infection and immunity ·Vol. 66 ·No. 11 ·1998-11-00 ·Pages 5301-6

Lay J, Henry LK, Clifford J, Koltin Y, Bulawa CE, Becker JM

Abstract

The ura-blaster technique for the disruption of Candida albicans genes has been employed in a number of studies to identify possible genes encoding virulence factors of this fungal pathogen. In this study, the URA3-encoded orotidine 5'-monophosphate (OMP) decarboxylase enzyme activities of C. albicans strains with ura-blaster-mediated genetic disruptions were measured. All strains harboring genetic lesions via the ura-blaster construct showed reduced OMP decarboxylase activities compared to that of the wild type when assayed. The activity levels in different gene disruptions varied, suggesting a positional effect on the level of gene expression. Because the URA3 gene of C. albicans has previously been identified as a virulence factor for this microorganism, our results suggest that decreased virulence observed in strains constructed with the ura-blaster cassette cannot accurately be attributed, in all cases, to the targeted genetic disruption. Although revised methods for validating a URA3-disrupted gene as a target for antifungal drug development could be devised, it is clearly desirable to replace URA3 with a different selectable marker that does not influence virulence.

MeSH Terms
Animals Candida albicans/enzymology,genetics,growth & development Enzyme Activation/genetics Fungal Proteins/biosynthesis,genetics Gene Deletion Gene Dosage Gene Expression Regulation, Fungal/genetics Mice Mutagenesis, Insertional Orotidine-5'-Phosphate Decarboxylase/genetics,metabolism Virulence/genetics
Chemicals
Fungal Proteins Orotidine-5'-Phosphate Decarboxylase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Lay J
Microbiology Department, University of Tennessee, Knoxville, Tennessee 37919, USA.
Henry L K
Clifford J
Koltin Y
Bulawa C E
Becker J M
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/IAI.66.11.5301-5306.1998
Published
1998-11-00
Pages
5301-6
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC108662
Subset
IM
External Links
PubMed source DOI: 10.1128/IAI.66.11.5301-5306.1998 PMC Full Text View journal details
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