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Antibiotics pp 227-235 | Cite as

Reporter Gene-Based Screening for TPP Riboswitch Activators

  • Christina E. Lünse
  • Günter Mayer
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1520)

Abstract

With the rise of multidrug resistant bacteria and a growing number of nosocomial infections, there has been an increased interest in finding new antibacterial drugs and drug targets. Riboswitches represent attractive new antibacterial drug targets, because they not only inherently recognize a specific metabolite or ion with their RNA aptamer domain, but also often regulate essential metabolic pathways. Here, we describe a reporter gene-based screen to identify compounds that activate the thiamine pyrophosphate (TPP) riboswitch in bacteria. This assay can be easily adapted for different riboswitch classes and thus has the potential to target many essential metabolic pathways and a broad spectrum of bacterial pathogens.

Key words

Translational fusion Miller assay Thiamine pyrophosphate lacZ reporter gene Antibiotics 

Notes

Acknowledgments

G.M. was supported by a grant from the German Research Foundation (MA3442/1-2). C.E.L. was supported by the German Research Foundation grant LU1889-1 and NIH P01 grant GM022778 awarded to Professor Ronald R. Breaker, HHMI investigator, Yale University.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Life and Medical Sciences InstituteUniversity of BonnBonnGermany
  2. 2.Department of Molecular, Cellular and Developmental BiologyYale UniversityNew HavenUSA

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