earticle

영어

Fluorescent protein variants with different spectral characteristics are highly desirable in broad range of biotechnology applications. Among them the green fluorescent protein (avGFP) from Aequorea victoria is unique fluorescent proteins and from the onset wild-type GFP, variety of mutant variants has been created and in many contexts they became a powerful reporter in the cellular and molecular biology. Several mutations were introduced at the chromophore and surrounding regions that resulted in various GFP mutants with different spectral properties which were successfully used in multi-color labeling, resonance energy transfer, and intracellular tracking studies. To obtain these variants, directed evolution and rational mutagenesis has been the method of choice to alter the spectral properties. Through these methods, only the canonical amino acids were used for replacing the chromophore residues of the protein. In particular, the presence of aromatic amino acid at Tyrosine 66 position is frequently used to create GFP variants. On the other hand, incorporation of non canonical amino acids into chromophore residues of GFP will overcome these limitations. The NCAA provides new functional groups to the protein that manipulates or enhances the spectral properties of the protein. Here, we reassigned of Tyrosine codon with different surrogates through selective pressure incorporation method. It offered different faces to the wild GFP with interesting features such as protein biosensor, protein conjugation, highly red shifted variant etc.