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Table 3 Properties of GFP derivatives used for setup of the mutational library and variants with improved in vivo expression in thermophilic bacilli

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

GFP Changes to A. victoria wild-type GFPa Properties Optimization method References
sfGFP(Sp) S30R, Y39N, F64L, S65T, Q80R, F99S, N105T, Y145F, M153T, V163A, I171V, A206V Used for random mutagenesis and FACS library construction in DSM 2542 in this study S. pneumoniae using OPTIMIZER program [35]
sfGFP(N39D/A179A) S30R, N39D, F64L, S65T, Q80R, F99S, N105T, Y145F, M153T, V163A, I171V, A179A, A206V Improved in vivo thermostability and brightness (885-fold) in comparison to sfGFP(Sp) at 60 °C Random mutagenesis and cell sorting by FACS This study
sfGFP(N39D/A179A/H231H) S30R, N39D, F64L, S65T, Q80R, F99S, N105T, Y145F, M153T, V163A, I171V, A179A, A206V, H231H Improved in vivo thermostability and brightness (310-fold) in comparison to sfGFP(Sp) at 60 °C Directed mutagenesis by introduction of silent mutation H231H into sfGFP(N39D/A179A) This study
sfGFP(N39D/Y66 W/A179A) S30R, N39D, F64L, S65T, Y66W, Q80R, F99S, N105T, Y145F, M153T, V163A, I171V, A179A, A206V Cyan variant with improved in vivo thermostability in thermophilic bacilli Directed mutagenesis by introduction of cyan color mutation Y66 W into sfGFP(N39D/A179A) This study
sfGFP(N39D/A179A/T203Y) S30R, N39D, F64L, S65T, Q80R, F99S, N105T, Y145F, M153T, V163A, I171V, A179A, T203Y, A206V Yellow variant with improved in vivo thermostability in thermophilic bacilli Directed mutagenesis by introduction of yellow color mutation T203Y into sfGFP(N39D/A179A) This study
  1. aItalicized amino acid exchanges indicate changes to the sfGFP(Sp) protein