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• W2024351785 abstract "The technique of repetitive extragenic palindromic polymerase chain reaction (REP-PCR) enables the identification and discrimination of clonally related Bacillus sporothermodurans isolates from ultra-high temperature and sterilized milk. The aim of this study was to investigate the genetic basis for the generation of these highly informative REP-PCR patterns. The major 947-bp REP-PCR fragment of B. sporothermodurans was cloned, together with its 5’ and 3’ flanking sequences. Only partial homology with the REP consensus sequence was established at the borders of the REP-PCR fragment. Moreover, these border sequences were located within two distinct open reading frames, with great homology to the uvrA and uvrB genes of Escherichia coli. The presence of these REP-like elements in B. sporothermodurans was thus sufficient for high resolution REP-PCR typing of this Gram-positive organism. In some cases (and especially with Gram-positives), REP-PCR could thus be considered more as an arbitrary primed PCR, albeit at a somewhat higher annealing temperature and using conserved primers. The random priming effect at the less stringent annealing conditions of REP-PCR was also demonstrated for enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) on another Gram-positive organism, Listeria monocytogenes. La présence d’éléments REP-like intragéniques est suffisante pour le typage PCR-REP. La PCR-REP assure l’identification et la distinction de souches du type Bacillus sporothermodurans isolées du lait stérilisé et notamment traité à hautes températures. L’objectif de la présente étude était d’analyser les bases génétiques pour l’élaboration de tels modèles PCR-REP hautement informatifs. Le fragment PCR-REP majeur, de 947 bp, de B. sporothermodurans a été cloné conjointement avec les séquences 5’ et 3’. Seule une homologie partielle, par rapport à la séquence REP consensus, est observée aux confins des fragments PCR-REP. En outre, ces séquences limites sont situées dans deux ORFs distincts, et l’on note une grande homologie avec les gènes uvrA et uvrB de Escherichia coli. La présence de ces éléments REP-like chez B. sporothermodurans est donc suffisant pour un typage PCR-REP décisif de cette bactérie Gram+. Dans certains cas (et tout particulièrement pour les Gram+), la PCR-REP peut être considérée notamment comme une PCR arbitraire quoique effectuée à une température élevée et avec des amorces conservées. La réaction d’amorçage au hasard, dans des conditions moins rigoureuses, a fait ses preuves également pour la PCR-ERIC dans la caractérisation d’une autre bactérie Gram+, Listeria monocytogenes." @default.
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• W2024351785 date "2000-05-01" @default.
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• W2024351785 title "The presence of intragenically located REP-like elements in Bacillus sporothermodurans is sufficient for REP-PCR typing" @default.
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• W2024351785 doi "https://doi.org/10.1016/s0923-2508(00)00146-7" @default.
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