Improved Green Fluorescent Protein by Molecular Evolution Using DNA Shuffling

  title={Improved Green Fluorescent Protein by Molecular Evolution Using DNA Shuffling},
  author={Andreas Crameri and Erik A. Whitehorn and Emily H Tate and Willem P. C. Stemmer},
  journal={Nature Biotechnology},
Green fluorescent protein (QFP) has rapidly become a widely used reporter of gene regulation. However, for many organisms, particularly eukaryotes, a stronger whole cell fluorescence signal is desirable. We constructed a synthetic GFP gene with improved codon usage and performed recursive cycles of DNA shuffling followed by screening for the brightest E. coli colonies. A visual screen using UV light, rather than FACS selection, was used to avoid red-shifting the excitation maximum. After 3… CONTINUE READING
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