PMID- 12732564
OWN - NLM
STAT- MEDLINE
DCOM- 20030805
LR  - 20210526
IS  - 0099-2240 (Print)
IS  - 1098-5336 (Electronic)
IS  - 0099-2240 (Linking)
VI  - 69
IP  - 5
DP  - 2003 May
TI  - In situ accessibility of Saccharomyces cerevisiae 26S rRNA to Cy3-labeled 
      oligonucleotide probes comprising the D1 and D2 domains.
PG  - 2899-905
AB  - Fluorescence in situ hybridization (FISH) has proven to be most useful for the 
      identification of microorganisms. However, species-specific oligonucleotide 
      probes often fail to give satisfactory results. Among the causes leading to low 
      hybridization signals is the reduced accessibility of the targeted rRNA site to 
      the oligonucleotide, mainly for structural reasons. In this study we used flow 
      cytometry to determine whole-cell fluorescence intensities with a set of 32 
      Cy3-labeled oligonucleotide probes covering the full length of the D1 and D2 
      domains in the 26S rRNA of Saccharomyces cerevisiae PYCC 4455(T). The brightest 
      signal was obtained with a probe complementary to positions 223 to 240. Almost 
      half of the probes conferred a fluorescence intensity above 60% of the maximum, 
      whereas only one probe could hardly detect the cells. The accessibility map based 
      on the results obtained can be extrapolated to other yeasts, as shown 
      experimentally with 27 additional species (14 ascomycetes and 13 basidiomycetes). 
      This work contributes to a more rational design of species-specific probes for 
      yeast identification and monitoring.
FAU - Inacio, Joao
AU  - Inacio J
AD  - Centro de Recursos Microbiologicos, Biotechnology Unit, Faculty of Sciences and 
      Technology, New University of Lisbon, 2829-516 Caparica, Portugal.
FAU - Behrens, Sebastian
AU  - Behrens S
FAU - Fuchs, Bernhard M
AU  - Fuchs BM
FAU - Fonseca, Alvaro
AU  - Fonseca A
FAU - Spencer-Martins, Isabel
AU  - Spencer-Martins I
FAU - Amann, Rudolf
AU  - Amann R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Appl Environ Microbiol
JT  - Applied and environmental microbiology
JID - 7605801
RN  - 0 (Carbocyanines)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Oligonucleotide Probes)
RN  - 0 (RNA, Fungal)
RN  - 0 (RNA, Ribosomal)
RN  - 0 (RNA, ribosomal, 26S)
RN  - 0 (cyanine dye 3)
SB  - IM
MH  - Base Sequence
MH  - Carbocyanines
MH  - Flow Cytometry
MH  - Fluorescent Dyes
MH  - In Situ Hybridization, Fluorescence
MH  - Nucleic Acid Conformation
MH  - Oligonucleotide Probes/chemistry/*genetics
MH  - RNA, Fungal/chemistry/*genetics
MH  - RNA, Ribosomal/chemistry/*genetics
MH  - Saccharomyces cerevisiae/*genetics
MH  - Species Specificity
PMC - PMC154520
EDAT- 2003/05/07 05:00
MHDA- 2003/08/06 05:00
PMCR- 2003/05/01
CRDT- 2003/05/07 05:00
PHST- 2003/05/07 05:00 [pubmed]
PHST- 2003/08/06 05:00 [medline]
PHST- 2003/05/07 05:00 [entrez]
PHST- 2003/05/01 00:00 [pmc-release]
AID - 1738 [pii]
AID - 10.1128/AEM.69.5.2899-2905.2003 [doi]
PST - ppublish
SO  - Appl Environ Microbiol. 2003 May;69(5):2899-905. doi: 
      10.1128/AEM.69.5.2899-2905.2003.