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

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.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Ghodke H,Caldas VE,Punter CM,van Oijen AM,Robinson A

doi

10.1016/j.bpj.2016.05.047

subject

Has Abstract

pub_date

2016-07-12 00:00:00

pages

25-7

issue

1

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(16)30394-0

journal_volume

111

pub_type

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