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Applied and Environmental Microbiology, October 2000, p. 4340-4344, Vol. 66, No. 10
Center for Food Safety and Quality
Enhancement, University of Georgia, Griffin, Georgia
30223-1797,1 and Department of
Microbiology and Biotechnology, University of Horticulture and Food
Science, Somloi ut 14-16, Budapest H-1118,
Hungary2
Received 13 March 2000/Accepted 28 July 2000
Yeast isolates from raw and processed poultry products were
characterized using PCR amplification of the internally transcribed spacer (ITS) 5.8S ribosomal DNA region (ITS-PCR), restriction analysis
of amplified products, randomly amplified polymorphic DNA (RAPD)
analysis, and pulsed-field gel electrophoresis (PFGE). ITS-PCR resulted
in single fragments of 350 and 650 bp, respectively, from eight strains
of Yarrowia lipolytica and seven strains of Candida
zeylanoides. Digestion of amplicons with HinfI and
HaeIII produced two fragments of 200 and 150 bp from
Y. lipolytica and three fragments of 350, 150, and 100 bp
from C. zeylanoides, respectively. Although these fragments
showed species-specific patterns and confirmed species identification,
characterization did not enable intraspecies typing. Contour-clamped
heterogeneous electric field PFGE separated chromosomal DNA of Y. lipolytica into three to five bands, most larger than 2 Mbp,
whereas six to eight bands in the range of 750 to 2,200 bp were
obtained from C. zeylanoides. Karyotypes of both yeasts
showed different polymorphic patterns among strains. RAPD analysis,
using enterobacterial repetitive intergenic sequences as primers,
discriminated between strains within the same species. Cluster analysis
of patterns formed groups that correlated with the source of isolation.
For ITS-PCR, extraction of DNA by boiling yeast cells was successfully used.
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Molecular Characterization of Yarrowia
lipolytica and Candida zeylanoides Isolated
from Poultry
*
Corresponding author. Mailing address: Center for Food
Safety and Quality Enhancement, 1109 Experiment St., Griffin, GA
30223-1797. Phone: (770) 412-4740. Fax: (770) 229-3216. E-mail:
lbeucha{at}cfsqe.griffin.peachnet.edu.
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