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DNA Repair (Amst). 2018 Mar;63:10-15. doi: 10.1016/j.dnarep.2018.01.006. Epub 2018 Jan 31.

Visualization of tandem repeat mutagenesis in Bacillus subtilis.

Author information

1
Department of General Microbiology, Georg-August University Göttingen, 37077, Göttingen, Germany.
2
Institute for Nonlinear Dynamics, Georg-August University Göttingen, 37077 Göttingen, Germany.
3
Institute of Bio and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, 52425 Jülich, Germany; RWTH Aachen University - Microscale Bioengineering (AVT.MSB), 52074 Aachen, Germany.
4
Multiscale Bioengineering, Bielefeld University, 33615 Bielefeld, Germany.
5
Department of General Microbiology, Georg-August University Göttingen, 37077, Göttingen, Germany. Electronic address: fcommic1@gwdg.de.

Abstract

Mutations are crucial for the emergence and evolution of proteins with novel functions, and thus for the diversity of life. Tandem repeats (TRs) are mutational hot spots that are present in the genomes of all organisms. Understanding the molecular mechanism underlying TR mutagenesis at the level of single cells requires the development of mutation reporter systems. Here, we present a mutation reporter system that is suitable to visualize mutagenesis of TRs occurring in single cells of the Gram-positive model bacterium Bacillus subtilis using microfluidic single-cell cultivation. The system allows measuring the elimination of TR units due to growth rate recovery. The cultivation of bacteria carrying the mutation reporter system in microfluidic chambers allowed us for the first time to visualize the emergence of a specific mutation at the level of single cells. The application of the mutation reporter system in combination with microfluidics might be helpful to elucidate the molecular mechanism underlying TR (in)stability in bacteria. Moreover, the mutation reporter system might be useful to assess whether mutations occur in response to nutrient starvation.

KEYWORDS:

Bacillus subtilis; Microfluidics; Mutagenesis; Mutational hot spot; Reporter system; Tandem repeat

PMID:
29414049
DOI:
10.1016/j.dnarep.2018.01.006
[Indexed for MEDLINE]

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