Affiliation:
1. Department of Microbiology and Institute of Cancer Research, Columbia University, New York, New York 10032
Abstract
ABSTRACT
Disruption of newly identified genes in the pathogen
Candida albicans
is a vital step in determination of gene function. Several gene disruption methods described previously employ long regions of homology flanking a selectable marker. Here, we describe disruption of
C. albicans
genes with PCR products that have 50 to 60 bp of homology to a genomic sequence on each end of a selectable marker. We used the method to disrupt two known genes,
ARG5
and
ADE2
, and two sequences newly identified through the
Candida
genome project,
HRM101
and
ENX3. HRM101
and
ENX3
are homologous to genes in the conserved
RIM101
(previously called
RIM1
) and
PacC
pathways of
Saccharomyces cerevisiae
and
Aspergillus nidulans
. We show that three independent
hrm101/hrm101
mutants and two independent
enx3/enx3
mutants are defective in filamentation on Spider medium. These observations argue that
HRM101
and
ENX3
sequences are indeed portions of genes and that the respective gene products have related functions.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
Reference24 articles.
1. A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae;Baudin A.;Nucleic Acids Res.,1993
2. Cloning and characterization of ECE1, a gene expressed in association with cell elongation of the dimorphic pathogen Candida albicans
3. Control of filament formation in Candida albicans by the transcriptional repressor TUP1;Braun B. R.;Science,1997
4. Candida albicans information. 19 December 1998 revision date. [Online.] University of Minnesota St. Paul.http://alces.med.umn.edu/Candida.html
[8 February 1999 last date accessed.]
5. Candida tblastn search. 21 October 1998 revision date. [Online.] Stanford University Palo Alto Calif.http://candida.stanford.edu/btComb.html
[8 February 1999 last date accessed.]