Premium
Ribosomal DNA spacer probes for yeast identification: Studies in the genus Metschnikowia
Author(s) -
Henriques Marília,
SáNogueira Isabel,
GiménezJurado Gabriella,
Van Uden N.
Publication year - 1991
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320070210
Subject(s) - biology , ribosomal dna , dna , spacer dna , schizosaccharomyces pombe , yarrowia , ribosomal rna , yeast , hybridization probe , strain (injury) , microbiology and biotechnology , internal transcribed spacer , nucleic acid thermodynamics , genetics , saccharomyces cerevisiae , gene , phylogenetics , base sequence , anatomy
To test whether DNA probes derived from ribosomal DNA spacer sequences are suitable for rapid and species‐specific yeast identification, a pilot study was undertaken. A 7·7 kb entire ribosomal DNA unit of the type strain of Metschnikowia reukaufii was isolated, cloned and mapped. A 0·65 kb Bam HI‐ Hpa I fragment containing nontranscribed spacer sequences was amplified and selected for testing as a 32 P hybridization probe with total DNA from the type strains of M. reukaufii , M. pulcherrima , M. lunata , M. bicuspidata , M. australis , M. zobellii , M. krissii , five other strains identified as M. reukaufii and strains of Schizosaccharomyces pombe , Hansenula canadensis , Saccharomyces cerevisiae and Yarrowia lipolytica . The probe hybridized exclsively with DNA from the type strain and four other strains of M. reukaufii . DNA from one strain labelled M. reukaufii did not hybridize with the probe. Subsequent % G+C comparison and DNA–DNA reassociation with the type strain revealed that the non‐hybridizing strain does not belong to the species M. reukaufii .