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 (K_d 120 nM, k_on 1.7 × 10⁵ M^-1 s^-1). In live E. coli cells, o

SUBMITTER: Eason MG 

PROVIDER: S-EPMC8477756 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature