Paulsson:Fluorescent proteins

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Brief description of what's out there and what we've got.

  • EYFP (is what people are usually referring to when they say just YFP)
GFP with S65G/S72A/T203Y
  • Citrine (Griesbeck JBC'01)
EYFP with V68L/Q69M
  • Venus (Nagai NBT'02)
EYFP with F46L/F64L/M153T/V163A/S175G
  • Our "Venus" (aka Citrus)
Venus with V68L/Q69M
  • Silver's Venus (Surprisingly, approx 8 fold less bright than Citrus when expressed in E.coli; expression may be the problem, rather than brightness)
Citrus with V68L / R80Q / H231L
  • Citrus-LAA
  • Citrus-LAA+2 (NY sequence repeated as spacer between ClpX and SspB binding sites)
  • weak RBS - Citrus
  • weak RBS - Citrus-LAA
  • mCherry


  • YPet (Nguyen NBT'05)

Venus with I47L/S208F/V224L/H231E/D234N

What mutations do

F46L: faster maturation at 37deg [1]
F64L: reduced sensitivity to pH and Cl-, folding
V68L: reduced sensitivity to pH [2]
Q69M: 2x photostabilty, lower pKa (5.7), indifference to chloride, better expression at 37deg and in organelles [3]
T203Y:
A206K: monomer (instead of weak dimer)

  1. Nagai T, Ibata K, Park ES, Kubota M, Mikoshiba K, and Miyawaki A. A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications. Nat Biotechnol. 2002 Jan;20(1):87-90. DOI:10.1038/nbt0102-87 | PubMed ID:11753368 | HubMed [Nagai]
  2. Miyawaki A, Griesbeck O, Heim R, and Tsien RY. Dynamic and quantitative Ca2+ measurements using improved cameleons. Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):2135-40. PubMed ID:10051607 | HubMed [Miyawaki]
  3. Griesbeck O, Baird GS, Campbell RE, Zacharias DA, and Tsien RY. Reducing the environmental sensitivity of yellow fluorescent protein. Mechanism and applications. J Biol Chem. 2001 Aug 3;276(31):29188-94. DOI:10.1074/jbc.M102815200 | PubMed ID:11387331 | HubMed [Griesbeck]
All Medline abstracts: PubMed | HubMed