Products/Services Used | Details | Operation |
---|---|---|
Gene Synthesis> | The genes encoding the wild-type yellow fluorescent protein from Phialidium sp. (phiYFP, UniProt ID: Q6RYS7) and the yellow fluorescent protein H148Q from Aequorea victoria (avYFP-H148Q, UniProt ID: P42212) were codon optimized for Escherichia coli K12 using the Integrated DNA Technologies Codon Optimization Tool. The genes were cloned into the pET-28b(+) vector between the Nde1 and BamH1 restriction sites with an N-terminal polyhistidine-tag and a C-terminal stop codon (Figure S23, GenScript, Piscataway, NJ). | Get A Quote |
Fluorescent proteins have been extensively engineered and applied as optical indicators for chloride in a variety of biological contexts. Surprisingly, given the biodiversity of fluorescent proteins, a naturally occurring chloride sensor has not been reported to date. Here, we present the identification and spectroscopic characterization of the yellow fluorescent protein from the jellyfish Phialidium sp . (phiYFP), a rare example of a naturally occurring, excitation ratiometric, and turn-on fluorescent protein sensor for chloride. Our results show that chloride binding tunes the p Ka of the chromophore Y66 and shifts the equilibrium from the fluorescent phenolate form to the weakly fluorescent phenol form. The ... More