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Codon Optimization> | … an N-terminal polyhistidine-tag and a C- terminal stop codon into the pET-28b(+) vector (Figure S20), GenScript, Piscataway, NJ) … The following day, the plasmids were isolated using the QIAprep Spin Miniprep Kit (catalog number: 27106; Qiagen, Hilden, Germany) and verified … | Get A Quote |
Gene Synthesis> | Get A Quote |
Chloride-sensitive fluorescent proteins generated from laboratory evolution have a characteristic tyrosine residue that interacts with a chloride ion and π-stacks with the chromophore. However, the engineered yellow-green fluorescent protein mNeonGreen lacks this interaction but still binds chloride, as seen in a recently reported crystal structure. Based on its unique coordination sphere, we were curious if chloride could influence the optical properties of mNeonGreen. Here, we present the structure-guided identification and spectroscopic characterization of mNeonGreen as a turn-on fluorescent protein sensor for chloride. Our results show that chloride binding lowers the chromophore pK and shifts the ... More