Yu JJ, Cole GT; American Society for Microbiology. General Meeting.
Abstr Gen Meet Am Soc Microbiol. 1997 May 4-8; 97: 263 (abstract no. F17).
Medical College of Ohio, Toledo, OH.
Coccidioides immitis is a fungal pathogen which causes a human respiratory disease known as San Joaquin Valley fever, or coccidioidomycosis. Identification of virulence factors of C. immitis ultimately requires confirmation by disruption of the putative gene which encodes a specific factor, followed by in vivo evaluation of altered pathogenicity of the disruptant. Orotidine monophosphate pyrophosphorylase (OMPPase), designated here as URA5, has been used successfully as a selective marker in several fungal gene transformation systems. In order to obtain this selective marker for developing a C. immitis transformation system, we set out to clone the ura5 gene by using a degenerate primer pair to two conserved regions of fungal URA5 proteins reported in the GenBank. A 135 base pair polymerase chain reaction (PCR) product was isolated and the translated sequence revealed 75% similarity to a reported Cryptococcus neoformans URA5. This PCR product was used to probe a C. immitis genomic library and a positive clone was selected and sequenced. The translated sequence of this putative ura5 gene showed 38.2-42.5% identity and 57.1-62.6% similarity to other reported fungal URA5 proteins. The ura5 gene of C. immitis is located on chromosome IV and has been determined to be a single gene, based on Southern hybridization of pulsed-field separated chromosomal DNA and restriction enzyme fragmented genomic DNA, respectively.
Publication Types:
Keywords:
- Antigens, Fungal
- Base Sequence
- Chromosomes
- Chromosomes, Fungal
- Coccidioides
- Coccidioidomycosis
- Cryptococcus neoformans
- Fungal Proteins
- Genes, Fungal
- Humans
- Polymerase Chain Reaction
- genetics
- immunology
- isolation & purification
Other ID:
UI: 102235329
From Meeting Abstracts