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Fluorescently Labeled DNA Interacts with Competence and Recombination Proteins and Is Integrated and Expressed Following Natural Transformation of Bacillus subtilis
Mirjam Boonstra, Nina Vesel, Oscar P. Kuipers
Department of Molecular Genetics, Faculty of Science and Engineering, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, Groningen, the Netherlands

Published: 25 September 2018 by American Society for Microbiology in mBio
American Society for Microbiology, Volume 9; 10.1128/mbio.01161-18
Abstract: We used DNA that was covalently labeled with fluorescent nucleotides to investigate the transformation process of Bacillus subtilis at the molecular level. We show that the labeled DNA colocalizes with components of the competence machinery, the chromosome, and the recombination protein RecA. Using time-lapse microscopy and microfluidics, we visualized, in real-time, the uptake of fluorescently labeled DNA. We found that under these conditions, cell division is not required for the expression of integrated DNA. Because the competence machinery is conserved in naturally competent bacteria, this method can also be used to investigate the transformation process in many other bacterial species.
Keywords: microfluidics, Transformation, Bacillus subtilis, Genetic Competence, labeled DNA
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