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Table 2 Properties of GFP derivatives used in this study for benchmarking the in vivo expression in P. thermoglucosidasius DSM 2542

From: In vivo selection of sfGFP variants with improved and reliable functionality in industrially important thermophilic bacteria

GFP Changes to A. victoria wild-type GFP Properties Gene/codon optimization method References
GFPuv aka “cycle3” mutant F99S, M153T, V163A Improved brightness by excitation with UV light Obtained by three cycles of DNA shuffling [81]
GFP+ F64L, S65T, Q80R, F99S, M153T, V163A 320-fold improved brightness in comparison to wild-type GFP Introduction of GFPmut1 mutation into GFPuv by PCR [55]
GFPmut3A S65G, S72A FACS-optimized mutant Obtained by random mutagenesis and cell sorting by FACS [82]
GFP(Sp) M1MV, S65A, V68L, S72A, A206K Codon-optimized Codon optimization for S. pneumoniae using OptimumGene software [83]
sfGFP S30R, Y39N, F64L, S65T, Q80R, F99S, N105T, Y145F, M153T, V163A, I171V, A206V Original superfolder GFP Obtained by DNA shuffling and fusion to poorly folding proteins. Based on the “folding reporter” protein that combined cycle3 (F99S, M153T, V163A) and enhanced GFP mutations (F64L, S65T) [38]
sfGFP(Gst) S30R, Y39N, F64L, S65T, Q80R, F99S, N105T, Y145F, M153T, V163A, I171V, A206V Codon-optimized superfolder GFP Codon optimization for Geobacillus stearothermophilus performed by GeneArt [40]
sfGFP(iGEM) S30R, Y39N, F64L, S65T, S72A, F99S, N105T, Y145F, M153T, V163A, I171V, A206V Codon-optimized superfolder GFP, additional GFPmut3* mutation (S2R) Codon optimization for E. coli and B. subtilis performed by GeneArt [84]
sfGFP(Bs) S30R, Y39N, F64L, S65T, Q80R, F99S, N105T, Y145F, M153T, V163A, I171V, A206V Codon-optimized superfolder GFP B. subtilis using dual codon method [35]
sfGFP(Sp) S30R, Y39N, F64L, S65T, Q80R, F99S, N105T, Y145F, M153T, V163A, I171V, A206V Codon-optimized superfolder GFP S. pneumoniae using OPTIMIZER program [35]