6533b85afe1ef96bd12b8cc6

RESEARCH PRODUCT

Fast protocols for the 5S rDNA and ITS-2 based identification ofOenococcus oeni

Inge SchönigJürgen FröhlichHelmut KönigArmin GneipelSteffen Hirschhäuser

subject

DNA BacterialMolecular Sequence DataDNA RibosomalPolymerase Chain ReactionMicrobiologyRibosome5S ribosomal RNASequence Homology Nucleic AcidDNA Ribosomal SpacerGeneticsmedicineInternal transcribed spacerMolecular BiologyGeneIn Situ Hybridization FluorescenceOenococcus oeniGeneticsBase Sequencebiologymedicine.diagnostic_testOligonucleotideRNA Ribosomal 5Sbiology.organism_classificationGram-Positive CocciRNA BacterialGenes BacterialNucleic Acid ConformationPrimer (molecular biology)LeuconostocFluorescence in situ hybridization

description

To identify specific marker sequences for the rapid identification of Oenococcus oeni, we sequenced the 23S-5S internal transcribed spacer (ITS-2) region and the 5S rDNA of five different O. oeni strains and three phylogenetically related lactic acid bacteria (LAB). Comparative analysis revealed 100% identity among the ITS-2 region of the O. oeni strains and remarkable differences in length and sequence compared to related LAB. These results enabled us to develop a primer set for a rapid PCR-identification of O. oeni within three hours. Moreover, the comparison of the 5S rDNA sequences and the highly conserved secondary structure provided the template for the design of three fluorescence-labeled specific oligonucleotides for fluorescence in situ hybridization (FISH). These probes are partial complementary to each other. This feature promotes the accessibility to the target sequence within the ribosome and enhances the fluorescence signal. For the rapid identification of Oenococci both the 5S rRNA gene and the ITS-2 region are useful targets.

yearjournalcountryeditionlanguage
2005-03-01FEMS Microbiology Letters
https://doi.org/10.1016/j.femsle.2005.01.033