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Clover

Clover is a basic (constitutively fluorescent) green/yellow fluorescent protein published in 2012, derived from Aequorea victoria. It is reported to be a rapidly-maturing weak dimer with high acid sensitivity.
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Oligomerization Organism Molecular Weight Cofactor
Weak dimer Aequorea victoria 26.8 kDa -

FPbase ID: 4Z641

Attributes

Ex λ Em λ EC (M-1 cm-1) QY Brightness pKa Maturation (min) Lifetime (ns)
505 515 111,000 0.76 84.36 6.2 30.0 3.2
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Clover OSER Measurements

% Normal Cells OSER/NE ratio Cell Type Reference
72.9 ± 5.5 (10000 cells) - HeLa Cranfill et al. (2016)
72.9 ± 5.5 (10000 cells) - HeLa Shaner et al. (2013)

Photostability

t1/2 (s) Power Light Mode In Cell Fusion ˚C Reference
50.0   Shaner et al. (2013)
A caution on interpretation of photostability measurements
Add photostability info

Clover Sequence

BLAST

Clover was derived from avGFP with the following mutations: M1_S2insV/S30R/Y39N/S65G/Q69A/F99S/N105T/Y145F/M153T/V163A/I171V/T203H

MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK
GenBank: AFR60231
IPG: 30275454

Structure

Deposited: ,
Chromophore:

Excerpts

With a completely deprotonated chromophore at neutral pH (pKa = 6.1), Clover should be incapable of photoconversion despite the presence of His203, which enhances photoconversion in PA-GFP. Indeed, we observed no photoactivation of Clover in illumination conditions that produced more than threefold photoactivation of PA-GFP.

Lam et al. (2012)

Primary Reference

Improving FRET dynamic range with bright green and red fluorescent proteins

Lam Aj, St-Pierre F, Gong Y, Marshall Jd, Cranfill Pj, Baird Ma, Mckeown Mr, Wiedenmann J, Davidson Mw, Schnitzer Mj, Tsien Ry, Lin Mz

(2012). Nature Methods, 9(10) , 1005-1012. doi: 10.1038/nmeth.2171. Article   Pubmed

Additional References

  1. Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range

    Leben R, Lindquist Rl, Hauser Ae, Niesner R, Rakhymzhan A

    (2022). International Journal of Molecular Sciences, 23(21) , 13407. doi: 10.3390/ijms232113407. Article

  2. Crystal Structure of Green Fluorescent Protein Clover and Design of Clover-Based Redox Sensors

    Campbell Bc, Petsko Ga, Liu Cf

    (2018). Structure, 26(2) , 225-237.e3. doi: 10.1016/j.str.2017.12.006. Article   Pubmed

  3. A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum

    Shaner Nc, Lambert Gg, Chammas A, Ni Y, Cranfill Pj, Baird Ma, Sell Br, Allen Jr, Day Rn, Israelsson M, Davidson Mw, Wang J

    (2013). Nature Methods, 10(5) , 407-409. doi: 10.1038/nmeth.2413. Article   Pubmed

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