Site-directed and random mutagenesis of mCerulean.
(a) Normalized absorption spectra of mCerulean and its mutants collected from sequential rounds of mutagenesis. All spectra were measured at pH 7.4 and 4°C. Black line – Cerulean, blue – V61K, cyan – V61K/D148G/Y151N, green – WasCFP (V61K/D148G/Y151N/L207Q) mutants. (b) Modeling of mutations V61K, D148G, Y151N and L207Q on Cerulean crystal structure (PDB ID: 2WSO) using PyMol package.