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dc.contributor.authorBarlag, Britta
dc.contributor.authorBeutel, Oliver
dc.contributor.authorJanning, Dennis
dc.contributor.authorCzarniak, Frederik
dc.contributor.authorRichter, Christian P
dc.contributor.authorKommnick, Carina
dc.contributor.authorGöser, Vera
dc.contributor.authorKurre, Rainer
dc.contributor.authorFabiani, Florian
dc.contributor.authorErhardt, Marc
dc.contributor.authorPiehler, Jacob
dc.contributor.authorHensel, Michael
dc.date.accessioned2016-09-13T09:40:47Z
dc.date.available2016-09-13T09:40:47Z
dc.date.issued2016
dc.identifier.citationSingle molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes. 2016, 6:31601 Sci Repen
dc.identifier.issn2045-2322
dc.identifier.pmid27534893
dc.identifier.doi10.1038/srep31601
dc.identifier.urihttp://hdl.handle.net/10033/620079
dc.description.abstractThe 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.
dc.language.isoenen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleSingle molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.en
dc.identifier.journalScientific reportsen
refterms.dateFOA2018-06-13T01:30:22Z
html.description.abstractThe 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.


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