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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

Faculty of Science, Medicine and Health - Papers: part A

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 …