GFP

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Contents

General Info

  • From jellyfish Aeqorea victoria
  • wtGFP sequence on NCBI
  • 238 amino acids
  • The fluorophore is tyrosine Y66 and the surrounding amino acids of Ser(65)-Tyr(66)-Gly(67) are also critical [1].

GFP Variants

  • GFPmut3b - S65G, S72A
  • GFPmut3* - mutations from wt: S2R,S65G,S72A
    • This is GFPmut3b[2] with a degredation tag attached + an accidental mutation at position 2 that doesn't affect function according to the authors.[3]
  • Emerald - S65T, S72A, N149K, M153T, I167T
  • EGFP - F64L, S65T + optimized for human codons with 35-fold increase in fluorescence over GFP [4]

Mutations

  • R96A: slows cyclization reaction from minutes to months
  • S65T: 5-6x increase in amplitude and red shift
  • Y203I: eliminate excitation peak at 475nm, leaving lower peak of 399nm. emission remains at 511nm, producing large Stokes shift
  • S65(G|T) and T203(Y|F|W|H) -> YFP
  • Y66W: GFP -> CFP
  • Y66H: GFP -> BFP (more blue than CFP), dim, easily photobleached
  • Y66F: excitation 360nm, emission 442nm

References

Error fetching PMID 9353317:
Error fetching PMID 8707053:
Error fetching PMID 9603842:
  1. Molecular Biology and Muation of Green Fluorescent Protein (Book Chapter, Zacharias & Tsien)

    [zacharias]

  2. Error fetching PMID 8707053: [Cormack]
  3. Error fetching PMID 9353317: [li97]
  4. Error fetching PMID 9603842: [Andersen]
All Medline abstracts: PubMed HubMed
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