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Single-molecule specific mislocalization of red fluorescent proteins in live Escherichia coli

Journal Article


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Abstract


  • Tagging of individual proteins with genetically encoded fluorescent proteins (FPs) has been used extensively to study localization and interactions in live cells. Recent developments in single-molecule localization microscopy have enabled the dynamic visualization of individual tagged proteins inside living cells. However, tagging proteins with FPs is not without problems: formation of insoluble aggregates and inhibition of native functions of the protein are well-known issues. Previously reported artifacts manifest themselves at all expression levels of the FP-tagged proteins, making the design of control experiments relatively straightforward. Here, we describe a previously uncharacterized mislocalization artifact of Entacmaea quadricolor red fluorescent protein variants that is detectable at the single-molecule level in live Escherichia coli cells.

Publication Date


  • 2016

Citation


  • Ghodke, H., Caldas, V. E. A., Punter, C. M., van Oijen, A. M. & Robinson, A. (2016). Single-molecule specific mislocalization of red fluorescent proteins in live Escherichia coli. Biophysical Journal, 111 (1), 25-27.

Scopus Eid


  • 2-s2.0-84994017728

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=5233&context=smhpapers

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/4209

Has Global Citation Frequency


Number Of Pages


  • 2

Start Page


  • 25

End Page


  • 27

Volume


  • 111

Issue


  • 1

Place Of Publication


  • United States

Abstract


  • Tagging of individual proteins with genetically encoded fluorescent proteins (FPs) has been used extensively to study localization and interactions in live cells. Recent developments in single-molecule localization microscopy have enabled the dynamic visualization of individual tagged proteins inside living cells. However, tagging proteins with FPs is not without problems: formation of insoluble aggregates and inhibition of native functions of the protein are well-known issues. Previously reported artifacts manifest themselves at all expression levels of the FP-tagged proteins, making the design of control experiments relatively straightforward. Here, we describe a previously uncharacterized mislocalization artifact of Entacmaea quadricolor red fluorescent protein variants that is detectable at the single-molecule level in live Escherichia coli cells.

Publication Date


  • 2016

Citation


  • Ghodke, H., Caldas, V. E. A., Punter, C. M., van Oijen, A. M. & Robinson, A. (2016). Single-molecule specific mislocalization of red fluorescent proteins in live Escherichia coli. Biophysical Journal, 111 (1), 25-27.

Scopus Eid


  • 2-s2.0-84994017728

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=5233&context=smhpapers

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/4209

Has Global Citation Frequency


Number Of Pages


  • 2

Start Page


  • 25

End Page


  • 27

Volume


  • 111

Issue


  • 1

Place Of Publication


  • United States