Titel:	Single molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes
Autor(en):	Barlag, Britta Beutel, Oliver Janning, Dennis Czarniak, Frederik Richter, Christian P. Kommnick, Carina Göser, Vera Kurre, Rainer Fabiani, Florian Erhardt, Marc Piehler, Jacob Hensel, Michael
Zusammenfassung:	The investigation of the subcellular localization, dynamics and interaction of proteins and protein complexes in prokaryotes is complicated by the small size of the cells. Super-resolution microscopy (SRM) comprise various new techniques that allow light microscopy with a resolution that can be up to ten-fold higher than conventional light microscopy. Application of SRM techniques to living prokaryotes demands the introduction of suitable fluorescent probes, usually by fusion of proteins of interest to fluorescent proteins with properties compatible to SRM. Here we describe an approach that is based on the genetically encoded self-labelling enzymes HaloTag and SNAP-tag. Proteins of interest are fused to HaloTag or SNAP-tag and cell permeable substrates can be labelled with various SRM-compatible fluorochromes. Fusions of the enzyme tags to subunits of a type I secretion system (T1SS), a T3SS, the flagellar rotor and a transcription factor were generated and analysed in living Salmonella enterica. The new approach is versatile in tagging proteins of interest in bacterial cells and allows to determine the number, relative subcellular localization and dynamics of protein complexes in living cells.
Bibliografische Angaben:	Scientific Reports, Vol. 6, Article number: 31601, 2016, S. 1-14
URL:	https://osnadocs.ub.uni-osnabrueck.de/handle/urn:nbn:de:gbv:700-2017021315524
Schlagworte:	Cellular microbiology; Super-resolution microscopy
Erscheinungsdatum:	13-Feb-2017
Lizenzbezeichnung:	Namensnennung 4.0 International
URL der Lizenz:	http://creativecommons.org/licenses/by/4.0/
Publikationstyp:	Einzelbeitrag in einer wissenschaftlichen Zeitschrift [article]
Enthalten in den Sammlungen:	FB05 - Hochschulschriften Open-Access-Publikationsfonds

Diese Ressource wurde unter folgender Copyright-Bestimmung veröffentlicht: Lizenz von Creative Commons