This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by McCullough, M. J.
Right arrow Articles by Stevens, D. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by McCullough, M. J.
Right arrow Articles by Stevens, D. A.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, April 1998, p. 1035-1038, Vol. 36, No. 4
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Intergenic Transcribed Spacer PCR Ribotyping for Differentiation of Saccharomyces Species and Interspecific Hybrids

Michael J. McCullough,1,2,3 Karl V. Clemons,1,2,3 John H. McCusker,4 and David A. Stevens1,2,3,*

Department of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford University School of Medicine, Stanford, California 943051; California Institute for Medical Research2 and Department of Medicine, Division of Infectious Diseases,3 Santa Clara Valley Medical Center, San Jose, California 95128; and Department of Microbiology, Duke University Medical Center, Durham, North Carolina 277104

Received 23 July 1997/Returned for modification 18 November 1997/Accepted 14 January 1998

The taxonomy of the genus Saccharomyces has undergone significant changes recently with the use of genotypic rather than phenotypic methods for the identification of strains to the species level. The sequence of rRNA genes has been utilized for the identification of a variety of fungi to the species level. This methodology, applied to species of Saccharomyces, allows unknown Saccharomyces isolates to be assigned to the type strains. It was the aim of the present study to assess whether typing of the intergenic spacer region by using restriction fragment length polymorphisms of PCR products (intergenic transcribed spacer PCR [ITS-PCR] ribotyping) could distinguish among type strains of the 10 accepted species of Saccharomyces and further to assess if this method could distinguish strains that were interspecific hybrids. Cellular DNA, isolated after the lysis of protoplasts, was amplified by PCR using ITS1 and ITS4 primers, purified by liquid chromatography, and digested with restriction endonucleases. Ribotyping patterns using the restriction enzymes MaeI and HaeIII could distinguish all species of Saccharomyces from each other, as well as from Candida glabrata, Candida albicans, and Blastomyces dermatitidis. The only exception to this was the inability to distinguish between Saccharomyces bayanus and S. pastorianus (S. carlsbergensis). Furthermore, interspecific hybrids resulting from the mating of sibling species of Saccharomyces were shown to share the ITS-PCR ribotyping patterns of both parental species. It should now be possible, by this simple PCR-based technique, to accurately identify these strains to the species level, thereby allowing an increase in our understanding of the characteristics required by these interspecific hybrids for their particular ecological niches.

* Corresponding author. Mailing address: Division of Infectious Diseases, Santa Clara Valley Medical Center, 751 South Bascom Ave., San Jose, CA 95128. Phone: (408) 885-4313. Fax: (408) 885-4306. E-mail: oflahren{at}wpgate.hhs.co.santa-clara.ca.us.

Journal of Clinical Microbiology, April 1998, p. 1035-1038, Vol. 36, No. 4
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Vollmer, T., Stormer, M., Kleesiek, K., Dreier, J. (2008). Evaluation of Novel Broad-Range Real-Time PCR Assay for Rapid Detection of Human Pathogenic Fungi in Various Clinical Specimens. J. Clin. Microbiol. 46: 1919-1926 [Abstract] [Full Text]  
  • Wang, Q.-M., Li, J., Wang, S.-A., Bai, F.-Y. (2008). Rapid Differentiation of Phenotypically Similar Yeast Species by Single-Strand Conformation Polymorphism Analysis of Ribosomal DNA. Appl. Environ. Microbiol. 74: 2604-2611 [Abstract] [Full Text]  
  • Bouchet, V., Huot, H., Goldstein, R. (2008). Molecular Genetic Basis of Ribotyping. Clin. Microbiol. Rev. 21: 262-273 [Abstract] [Full Text]  
  • Guimaraes, P. M. R., Francois, J., Parrou, J. L., Teixeira, J. A., Domingues, L. (2008). Adaptive Evolution of a Lactose-Consuming Saccharomyces cerevisiae Recombinant. Appl. Environ. Microbiol. 74: 1748-1756 [Abstract] [Full Text]  
  • Gerke, J. P., Chen, C. T. L., Cohen, B. A. (2006). Natural Isolates of Saccharomyces cerevisiae Display Complex Genetic Variation in Sporulation Efficiency. Genetics 174: 985-997 [Abstract] [Full Text]  
  • Mentel, M., Spirek, M., Jorck-Ramberg, D., Piskur, J. (2006). Transfer of Genetic Material between Pathogenic and Food-Borne Yeasts.. Appl. Environ. Microbiol. 72: 5122-5125 [Abstract] [Full Text]  
  • Rainieri, S., Kodama, Y., Kaneko, Y., Mikata, K., Nakao, Y., Ashikari, T. (2006). Pure and Mixed Genetic Lines of Saccharomyces bayanus and Saccharomyces pastorianus and Their Contribution to the Lager Brewing Strain Genome.. Appl. Environ. Microbiol. 72: 3968-3974 [Abstract] [Full Text]  
  • Ahmed, A. O. A., Mukhtar, M. M., Kools-Sijmons, M., Fahal, A. H., de Hoog, S., van den Ende, B. G., Zijlstra, E. E., Verbrugh, H., Abugroun, E. S. A. M., Elhassan, A. M., van Belkum, A. (1999). Development of a Species-Specific PCR-Restriction Fragment Length Polymorphism Analysis Procedure for Identification of Madurella mycetomatis. J. Clin. Microbiol. 37: 3175-3178 [Abstract] [Full Text]  
  • Posteraro, B., Sanguinetti, M., D'Amore, G., Masucci, L., Morace, G., Fadda, G. (1999). Molecular and Epidemiological Characterization of Vaginal Saccharomyces cerevisiae Isolates. J. Clin. Microbiol. 37: 2230-2235 [Abstract] [Full Text]  
  • McCullough, M. J., Clemons, K. V., McCusker, J. H., Stevens, D. A. (1998). Species Identification and Virulence Attributes of Saccharomyces boulardii (nom. inval.). J. Clin. Microbiol. 36: 2613-2617 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»