Validation of a fluorescence-based screening concept to identify ribosome assembly defects in Escherichia coli.

ABSTRACT: While the structure of mature ribosomes is analyzed in atomic detail considerably less is known about their assembly process in living cells. This is mainly due to technical and conceptual hurdles. To analyze ribosome assembly in vivo, we designed and engineered an Escherichiacoli strain--using chromosomal gene knock-in techniques--that harbors large and small ribosomal subunits labeled with the fluorescent proteins EGFP and mCherry, respectively. A thorough characterization of this reporter strain revealed that its growth properties and translation apparatus were wild-type like. Alterations in the ratio of EGFP over mCherry fluorescence are supposed to indicate ribosome assembly defects. To provide proof of principle, subunit specific assembly defects were provoked and could be identified

SUBMITTER: Nikolay R 

PROVIDER: S-EPMC4081057 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature