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Updated: Apr 12, 2026
Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
Published on: January 6, 2015
Developing a genetically encoded green fluorescent protein mutant for sensitive light-up fluorescent sensing and
Tao Jiang1, Daiping Guo1, Qian Wang1
1Department of Chemistry, College of Chemistry and Chemical Engineering, the MOE Key Laboratory of Analytical Sciences, Xiamen University, Xiamen 361005, China.
Abstract:
Hg(II) is well-known for quenching fluorescence in a distance dependent manner. Nevertheless, when we exposed the fluorophore of a green fluorescent protein (GFP) toward Hg(II), through H148C mutation, the GFP fluorescence could be "lighted up" by Hg(II) down to sub-nM level. The detection linear range is 0.5-3.0 nM for protein solutions at 8.0 nM. The GFPH148C protein displayed a promising selectivity toward Hg(II) and also the cellular imaging capacity. Spectra measurements suggested that the ground-state redistribution of protein contributed to the fluorescence enhancement, which was found not limited to Hg(II), and thus presented an opening for building a pool of GFP-based chemosensors toward other heavy metal ions.
