Evolutionary optimization of fluorescent proteins for intracellular FRET

Nguyen Aw, Daugherty Ps

(2005). Nature Biotechnology, 23(3) , 355-360. doi: 10.1038/nbt1066. Article   Pubmed

    Primary Proteins:
  1. CFP4
  2. CyPet
  3. YFP3
  4. YPet
Add photostability measurements

Excerpts

YFP3 possessed six mutations that resulted in an RRC almost fivefold greater than that of the parental pair. YFP3 also conferred a roughly threefold improvement in FRET relative to the fast-maturing Venus YFP. In an attempt to further enhance YFP3 brightness, the Venus mutations were combined with those of YFP3 to yield YFP for energy transfer (YPet). Though YFP3 exhibited slower folding relative to Venus, combining YFP3 and Venus mutations resulted in intermediate folding kinetics.

YFP3 possessed six mutations that resulted in an RRC almost fivefold greater than that of the parental pair. YFP3 also conferred a roughly threefold improvement in FRET relative to the fast-maturing Venus YFP.