Abstract
A cloned DNA fragment from Candida albicans containing the gene for the protein actin was used to probe the molecular structure of the actin gene of several medically important yeasts (C. albicans, Candida stellatoidea, Candida tropicalis, Candida pseudotropicalis, Candida krusei, Candida parapsilosis, Candida guilliermondii, and Torulopsis glabrata). Whole-cell DNA from each species was digested with restriction endonucleases, electrophoresed on agarose gels, and transferred to nitrocellulose. Radioactively labeled C. albicans actin gene was hybridized to the DNA fragments on the nitrocellulose. The C. albicans probe produced a strong signal with all of the Candida DNAs tested, indicating considerable conservation of this gene. In addition, the actin genes of all of the species tested were found to have no internal EcoRI or SalI restriction sites. With the exception of C. guilliermondii, all of the species tested had a single internal HindIII recognition site. However, the location of flanking restriction sites was found to be species specific. For all of the enzymes tested, the locations of the flanking restriction sites in C. albicans and C. stellatoidea were identical; all of the other strains yielded fragments clearly distinct from one another. These differences provide a molecular tool for the differentiation of medically important Candida species.
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- Brawner D. L., Cutler J. E. Variability in expression of a cell surface determinant on Candida albicans as evidenced by an agglutinating monoclonal antibody. Infect Immun. 1984 Mar;43(3):966–972. doi: 10.1128/iai.43.3.966-972.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Langer P. R., Waldrop A. A., Ward D. C. Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6633–6637. doi: 10.1073/pnas.78.11.6633. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leary J. J., Brigati D. J., Ward D. C. Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4045–4049. doi: 10.1073/pnas.80.13.4045. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McGinnis M. R., Ajello L., Beneke E. S., Drouhet E., Goodman N. L., Halde C. J., Haley L. D., Kane J., Land G. A., Padhye A. A. Taxonomic and nomenclatural evaluation of the genera Candida and Torulopsis. J Clin Microbiol. 1984 Oct;20(4):813–814. doi: 10.1128/jcm.20.4.813-814.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Norgard M. V. Rapid and simple removal of contaminating RNA from plasmid DNA without the use of RNase. Anal Biochem. 1981 May 1;113(1):34–42. doi: 10.1016/0003-2697(81)90040-3. [DOI] [PubMed] [Google Scholar]
- O'Farrell P. H., Kutter E., Nakanishi M. A restriction map of the bacteriophage T4 genome. Mol Gen Genet. 1980;179(2):421–435. doi: 10.1007/BF00425473. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Riggsby W. S., Torres-Bauza L. J., Wills J. W., Townes T. M. DNA content, kinetic complexity, and the ploidy question in Candida albicans. Mol Cell Biol. 1982 Jul;2(7):853–862. doi: 10.1128/mcb.2.7.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Syverson R. E. Variable assimilation of carbon compounds by Candida albicans. J Clin Microbiol. 1981 Jan;13(1):163–166. doi: 10.1128/jcm.13.1.163-166.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Torres-Bauzá L. J., Riggsby W. S. Protoplasts from yeast and mycelial forms of Candida albicans. J Gen Microbiol. 1980 Aug;119(2):341–349. doi: 10.1099/00221287-119-2-341. [DOI] [PubMed] [Google Scholar]
- Wills J. W., Lasker B. A., Sirotkin K., Riggsby W. S. Repetitive DNA of Candida albicans: nuclear and mitochondrial components. J Bacteriol. 1984 Mar;157(3):918–924. doi: 10.1128/jb.157.3.918-924.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wills J. W., Troutman W. B., Riggsby W. S. Circular mitochondrial genome of Candida albicans contains a large inverted duplication. J Bacteriol. 1985 Oct;164(1):7–13. doi: 10.1128/jb.164.1.7-13.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zeman L. J., Lusena C. V. In vivo and in vitro synthesis of yeast mitochondrial DNA. Methods Cell Biol. 1975;12:273–283. doi: 10.1016/s0091-679x(08)60960-7. [DOI] [PubMed] [Google Scholar]