Very bright green fluorescent proteins from the Pontellid copepod Pontella mimocerami.

PloS One
Marguerite E HuntMikhail V Matz

Abstract

Fluorescent proteins (FP) homologous to the green fluorescent protein (GFP) from the jellyfish Aequorea victoria have revolutionized biomedical research due to their usefulness as genetically encoded fluorescent labels. Fluorescent proteins from copepods are particularly promising due to their high brightness and rapid fluorescence development. Here we report two novel FPs from Pontella mimocerami (Copepoda, Calanoida, Pontellidae), which were identified via fluorescence screening of a bacterial cDNA expression library prepared from the whole-body total RNA of the animal. The proteins are very similar in sequence and spectroscopic properties. They possess high molar extinction coefficients (79,000 M(-1) cm(-)) and quantum yields (0.92), which make them more than two-fold brighter than the most common FP marker, EGFP. Both proteins form oligomers, which we were able to counteract to some extent by mutagenesis of the N-terminal region; however, this particular modification resulted in substantial drop in brightness. The spectroscopic characteristics of the two P. mimocerami proteins place them among the brightest green FPs ever described. These proteins may therefore become valuable additions to the in vivo imaging toolkit.

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Citations

Jun 16, 2014·Nature Methods·Alfredo Erazo-OliverasJean-Philippe Pellois
May 27, 2011·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Galina V AglyamovaMikhail V Matz
Jan 19, 2012·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Marguerite E HuntMikhail V Matz
Dec 21, 2013·Biotechnology Journal·Fabienne MérolaHélène Pasquier
Aug 26, 2016·BMC Evolutionary Biology·Warren R FrancisSteven H D Haddock
Jun 17, 2020·Zoological Letters·Marie-Lyne MacelSalvatore D'Aniello
Dec 29, 2017·The Journal of Biological Chemistry·Melanie J BlandenJames L Hougland

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Datasets Mentioned

BETA
GQ247522
GQ247523

Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

MrBayes
Geneious package
BLASTX
Geneious
Image J
SeqMan2
Expressionist
OpenLab
DNASTAR Lasergene

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