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 |
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. DOI:10.1073/pnas.96.5.2135 |
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 |