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].
  • Oxygen is required for maturation of the fluorophore

GFP Variants

  • GFPmut3b - S65G, S72A
  • GFPmut3* - mutations from wt: S2R,S65G,S72A
    • This is GFPmut3b[2] with an accidental mutation at position 2 that doesn't affect function according to the authors.[3]
  • Emerald - F64L, S65T, S72A, N149K, M153T, I167T
  • EGFP - added GTG as second codon, F64L, S65T + optimized for human codons with 35-fold increase in fluorescence over GFP [4]
  • Other variants and their mutations can be found in Shaner supplementary table 2 [5]

Mutations

  • F64L: improve folding @37C
  • S65T: 5-6x increase in amplitude and red 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
  • R96A: slows cyclization reaction from minutes to months
  • Y203I: eliminate excitation peak at 475nm, leaving lower peak of 399nm. emission remains at 511nm, producing large Stokes shift
  • A206K: make monomeric

References

General

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  1. Molecular Biology and Muation of Green Fluorescent Protein (Book Chapter, Zacharias & Tsien)

    [zacharias]

  2. Error fetching PMID 8707053: [Cormack]
  3. Error fetching PMID 9603842: [Andersen]
  4. Error fetching PMID 9353317: [li97]
  5. Error fetching PMID 16299475: [shaner05]
  6. Error fetching PMID 1347277: [prasher]
    original cloning of GFP

  7. Error fetching PMID 8303295: [chalfie]
    original use of GFP as a reporter

  8. Error fetching PMID 9759496: [tsien98]
    good review of GFP

All Medline abstracts: PubMed HubMed

GFP as a measure of gene expression

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  1. Error fetching PMID 11698362: [Lindow]
  2. Error fetching PMID 18350571: [Iafolla]
  3. Error fetching PMID 18352063: [Bagh]
  4. Error fetching PMID 18352052: [Dong]
All Medline abstracts: PubMed HubMed
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