Single-Molecule Specific Mislocalization of Red Fluorescent Proteins in Live Escherichia coli

Harshad Ghodke, Victor E. A. Caldas, Christiaan M. Punter, Antoine M. van Oijen*, Andrew Robinson

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

8 Citations (Scopus)
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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 quadri-color red fluorescent protein variants that is detectable at the single-molecule level in live Escherichia coli cells.

Original languageEnglish
Pages (from-to)25-27
Number of pages3
JournalBiophysical Journal
Volume111
Issue number1
DOIs
Publication statusPublished - 12-Jul-2016

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