GFP

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 zacharias.
 * Oxygen is required for maturation of the fluorophore

GFP Variants

 * GFPmut3b - S65G, S72A
 * GFPmut3* - mutations from wt: S2R,S65G,S72A
 * This is GFPmut3b Cormack with an accidental mutation at position 2 that doesn't affect function according to the authors. Andersen
 * 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 li97
 * Other variants and their mutations can be found in Shaner supplementary table 2 shaner05

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

General
//original cloning of GFP //original use of GFP as a reporter //good review of GFP
 * 1) prasher pmid=1347277
 * 1) chalfie pmid=8303295
 * 1) tsien98 pmid=9759496
 * 1) li97 pmid=9353317
 * 2) Cormack pmid=8707053
 * 3) Andersen pmid=9603842
 * 4) zacharias Molecular Biology and Muation of Green Fluorescent Protein (Book Chapter, Zacharias & Tsien)
 * 5) shaner05 pmid=16299475

GFP as a measure of gene expression

 * 1) Lindow pmid=11698362
 * 2) Iafolla pmid=18350571
 * 3) Bagh pmid=18352063
 * 4) Dong pmid=18352052