Genetically Encoded Fluorescent Biosensor for Rapid Detection of Protein Expression.

Abstract:

:Fluorescent proteins are widely used as fusion tags to detect protein expression in vivo. To become fluorescent, these proteins must undergo chromophore maturation, a slow process with a half-time of 5 to >30 min that causes delays in real-time detection of protein expression. Here, we engineer a genetically encoded fluorescent biosensor to enable detection of protein expression within seconds in live bacteria. This sensor for transiently expressed proteins (STEP) is based on a fully matured but dim green fluorescent protein in which pre-existing fluorescence increases 11-fold in vivo following the specific and rapid binding of a protein tag (Kd 120 nM, kon 1.7 × 105 M-1 s-1). In live E. coli cells, our STEP biosensor enables detection of protein expression twice as fast as the use of standard fluorescent protein fusions. Our biosensor opens the door to the real-time study of short timescale processes in live cells with high spatiotemporal resolution.

journal_name

ACS Synth Biol

journal_title

ACS synthetic biology

authors

Eason MG,Pandelieva AT,Mayer MM,Khan ST,Garcia HG,Chica RA

doi

10.1021/acssynbio.0c00407

subject

Has Abstract

pub_date

2020-11-20 00:00:00

pages

2955-2963

issue

11

issn

2161-5063

journal_volume

9

pub_type

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