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Maniatis T: Molecular Cloning: A Laboratory Manual. 2nd edition. Cold Spring Habor, NY: Cold Spring Habor Laboratory Press; 1989. 26. McLaughlan AM, Foster J: Molecular characterization of an Selleckchem Gilteritinib autolytic amidase of Listeria monocytogenes EGD. Microbiology 1998, 144:1359–1367.PubMedCrossRef 27. Park SF, Stewart GS: High-efficiency transformation of Listeria monocytogenes by electroporation of penicillin-treated cells. Gene 1990, 94:129–132.PubMedCrossRef Authors’ contributions AK-B carried out the molecular cloning to create the constructs to apply the NICE system in L. monocytogenes, performed the analysis of PBPs as
well as the susceptibility studies, and helped to draft the manuscript. MP carried out the studies on growth and cell morphology of the obtained recombinant strains. ZM conceived part of the study, participated in its design and coordinated the preparation of the manuscript. VX-765 in vitro All authors read and approved the final version of the manuscript.”
“Background mTOR inhibitor Scientists today are studying bacterial communities from diverse habitats, hosts, and health conditions based on the 16 S rRNA gene [1, 2]. To date, most studies have focused on qualitative characterization based on the relative abundances of community bacterial groups [3–5]; however, quantitative characterization—i.e., measurement of the total
bacterial load—provides valuable and complementary information when combined with these qualitative data [6]. Traditional culture-based approaches for quantifying bacterial load are inherently limited for assessing the complex bacterial communities that exist in many clinical and environmental samples. Likewise, standard culture-based methods are ineffective for quantifying many fastidious and uncultivable bacterial species [7]. Among culture-independent approaches, quantitative real-time PCR (qPCR) is currently best suited for measuring bacterial load, because of its intrinsic quantitative capability, ease of use, and flexibility in assay design [8, 9]. Using the qPCR platform, we can design an assay capable of concurrently detecting and quantifying all unique bacteria that constitutes a complex community.