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PMID: 10074081 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Rapid hypothesis testing with Candida albicans through gene disruption with short homology regions.

Journal of bacteriology ·Vol. 181 ·No. 6 ·1999-03-00 ·Pages 1868-74

Wilson RB, Davis D, Mitchell AP

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.

MeSH Terms
Alleles Base Sequence Candida albicans/genetics,pathogenicity,physiology DNA Primers/genetics DNA, Fungal/genetics Genes, Fungal Genetic Markers Genetic Techniques Molecular Sequence Data Mutagenesis Mutation Phenotype Polymerase Chain Reaction Restriction Mapping Transformation, Genetic
Chemicals
DNA Primers DNA, Fungal Genetic Markers
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Wilson R B
Department of Microbiology and Institute of Cancer Research, Columbia University, New York, New York 10032, USA.
Davis D
Mitchell A P
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.181.6.1868-1874.1999
Published
1999-03-00
Pages
1868-74
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC93587
Subset
IM
Grants
NIAID NIH HHS · R01 AI070272 · United States
NIAID NIH HHS · T32 AI007161 · United States
NIAID NIH HHS · T32 AI07161-21 · United States
Databases
GENBANK
AF173954, AF173955, AF173956
External Links
PubMed source DOI: 10.1128/JB.181.6.1868-1874.1999 PMC Full Text View journal details
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