Candida albicans is an important human pathogen closely related to the intensively studied yeast, Saccharomyces cerevisiae. C. albicans causes a range of illnesses, from oral thrush in infants and vaginitis in women, to severe and life-threatening systemic infections in patients with compromised immune systems. The limited number of effective and safe antifungal antibiotics underscores the importance of increasing our understanding of the unique genetic traits of C. albicans. A shotgun genomic sequencing approach will be utilized to obtain minimal coverage of the genome and thereby to identify most of the genes from this pathogenic yeast. The goals of this project are: 1) to obtain approximately 1.5X sequence coverage of the 16Mb Candida albicans genome; 2) to obtain at least partial sequence data for greater than 95 percent of the encoded genes; 3) to identify a subset of the encoded genes which may have interesting functions, such as mating type loci, drug resistance markers, and pathogenicity genes, and to complete the sequence of these genes at high accuracy; and 4) to establish a database of all C. albicans sequence data, trace files, predicted genes, and BLAST search results, integrated with the current C albicans WWW server. Genomic DNA from C. albicans will be randomly sheared and cloned into a plasmid vector. Cycle sequencing reactions will be performed using fluorescent primer chemistry from both ends of each insert. Sequence analysis will be carried out on an ABI 377 Sequencer, and data will be transferred automatically to a UNIX system. The complete sequence of interesting genes will be carried out by primer walking with fluorescent terminators to complete the sequence of the original plasmid clone; in most cases, it will also be necessary to use this clone to probe the reference large-insert genomic library to identify larger clones which will contain the ends of the genes. A new database will be constructed to integrate the clone information, raw sequence and trace data, and the results of homology searches. This genomic sequencing approach is essentially a large-scale method of gene discovery which will quickly and economically provide key research tools to the entire Candida albicans community.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
3R01DE012302-02S1
Application #
2656026
Study Section
Special Emphasis Panel (ZRG2-GNM (03))
Project Start
1996-09-30
Project End
2000-07-31
Budget Start
1997-08-01
Budget End
2000-07-31
Support Year
2
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Stanford University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
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