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Infection and Immunity, November 1998, p. 5301-5306, Vol. 66, No. 11
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Altered Expression of Selectable Marker URA3 in Gene-Disrupted Candida albicans Strains Complicates Interpretation of Virulence Studies

Jennifer Lay,1 L. Keith Henry,1 Julie Clifford,2 Yigal Koltin,2 Christine E. Bulawa,2 and Jeffrey M. Becker1,*

Microbiology Department, University of Tennessee, Knoxville, Tennessee 379191 and Millennium Pharmaceuticals, Inc., Cambridge, Massachusetts 021392

Received 17 April 1998/Returned for modification 3 June 1998/Accepted 1 September 1998

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.

* Corresponding author. Mailing address: Microbiology Department, University of Tennessee, F339 Walters Life Sciences Bldg., Knoxville, TN 37996-0845. Phone: (423) 974-3006. Fax: (423) 974-4007. E-mail: jbecker{at}utk.edu.

Infection and Immunity, November 1998, p. 5301-5306, Vol. 66, No. 11
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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