GFP
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*[http://parts.mit.edu/registry/index.php/Part:BBa_E0040 ''GFP''mut3*] - mutations from wt: S2R,S65G,S72A | *[http://parts.mit.edu/registry/index.php/Part:BBa_E0040 ''GFP''mut3*] - mutations from wt: S2R,S65G,S72A | ||
**This is GFPmut3b<cite>Cormack</cite> with a degredation tag attached + an accidental mutation at position 2 that doesn't affect function according to the authors.<cite>Anderson</cite> | **This is GFPmut3b<cite>Cormack</cite> with a degredation tag attached + an accidental mutation at position 2 that doesn't affect function according to the authors.<cite>Anderson</cite> | ||
| - | *[[Emerald]] | + | *[[Emerald]] - S65T, S72A, N149K, M153T, I167T |
| - | * EGFP F64L, S65T + optimized for human codons with 35-fold increase in fluorescence over GFP <cite>li97</cite> | + | * EGFP - F64L, S65T + optimized for human codons with 35-fold increase in fluorescence over GFP <cite>li97</cite> |
==Mutations== | ==Mutations== | ||
Revision as of 15:20, 27 September 2006
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
- 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
- Molecular Biology and Muation of Green Fluorescent Protein (Book Chapter, Zacharias & Tsien)
- Cormack BP, Valdivia RH, and Falkow S. . pmid:8707053.
- Li X, Zhang G, Ngo N, Zhao X, Kain SR, and Huang CC. . pmid:9353317.
- Andersen JB, Sternberg C, Poulsen LK, Bjorn SP, Givskov M, and Molin S. . pmid:9603842.
