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Article
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The threonine-tRNA ligase gene region is applicable in classification, typing, and phylogenetic analysis of bifidobacteria
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Ji 345;í Killer1,2, Chahrazed Mekadim 1,2, Radko Pechar 2,3, V 283;ra Bunešová, Eva Vlková 2
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Journal of Microbiology 2018;56(10):713-721.
DOI: https://doi.org/10.1007/s12275-018-8167-3
Published online: September 28, 2018
1Institute of Animal Physiology and Genetics of the Czech Academy of Sciences, Vídeňská 1083, 142 20, Czechia, 2Czech University of Life Sciences, Faculty of Agrobiology, Food and Natural Resources, Department of Microbiology, Nutrition and Dietetics, Kamýcká 129, 165 00, Czechia, 3Food Research Institute Prague, Radiová 1285/7, 102 00, Czechia
Received: 27 March 2018 • Revised: 26 June 2018 • Accepted: 27 June 2018
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Abstract
In the modern era, molecular genetic techniques are crucial
in ecological studies, as well as in the classification, typing,
and phylogenetic analysis of prokaryotes. These techniques
are mainly aimed at whole genome comparisons and PCRderived
experiments, including amplifying the 16S rRNA
and other various housekeeping genes used in taxonomy,
as well as MLST (multilocus sequence typing) and MLSA
(multilocus sequence analysis) of different taxonomic bacterial
groups. The gene encoding threonine-tRNA ligase
(thrS) is a gene potentially applicable as an identification
and phylogenetic marker in bacteria. It is widely distributed
in bacterial genomes and is subject to evolutionary selection
pressure due to its important function in protein synthesis.
In this study, specific primers were used to amplify a thrS
gene fragment (~740 bp) in 36 type and 30 wild strains classified
under family Bifidobacteriaceae. The full-length gene
has not yet been considered as a possible identification, classification,
and phylogenetic marker in bifidobacteria. The
thrS sequences revealed higher sequence variability (82.7%
of pairwise identities) among members of the family than
that shown by 16S rRNA gene sequences (96.0%). Although
discrepancies were found between the thrS-derived and previously
reported whole genome phylogenetic analyses, the
main phylogenetic groups of bifidobacteria were properly
assigned. Most wild strains of bifidobacteria were better differentiated
based on their thrS sequences than on their 16S
rRNA gene identities. Phylogenetic confidence of the evaluated
gene with respect to other alternative genetic markers
widely used in taxonomy of bifidobacteria (fusA, GroELhsp60,
pyrG, and rplB genes) was confirmed using the localized
incongruence difference - Templeton analysis.
Supplementary Information
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