Yellow fluorescent protein TurboYFP

- Superbright true-yellow fluorescence
- Fast maturation
- Emission wavelength is ideally positioned between those of green and red fluorescent proteins
- Destabilized version is available
- Recommended for gene expression analysis, cell and organelle labeling

TurboYFP is an enhanced variant of the yellow fluorescent protein PhiYFP from jellyfish Phialidium sp. [Shagin et al., 2004]. Possessing superbright yellow fluorescence with emission maximum at 538 nm, TurboYFP is ideally positioned between green and red fluorescent proteins, allowing easy separation of these fluorescent markers by flow cytometry using common channels of detection and a single laser excitation line. Compared with Phi-Yellow proteins, TurboYFP matures faster in mammalian cells. TurboYFP is mainly intended for applications where fast appearance of bright fluorescence is crucial. It is specially recommended for cell labeling and tracking the promoter activity. Destabilized TurboYFP variant allows accurate analysis of rapid and/or transient events in gene regulation.

Main properties

TurboYFP normalized excitation (thin line) and emission (thick line) spectra. Download TurboYFP spectra (xls)

CHARACTERISTIC *Brightness is a product of extinction coefficient and quantum yield, divided by 1000. Molecular weight, kDa 26 Polypeptide length, aa 234 Fluorescence color yellow Excitation maximum, nm 525 Emission maximum, nm 538 Quantum yield 0.53 Extinction coefficient, M-1cm-1 105 000 Brightness* 55.7 Brightness, % of EGFP 169 pKa 5.9 Structure dimer Aggregation at high concentrations Maturation rate at 37°C super fast Photostability high Cell toxicity at high concentrations Main advantages superbright and fast-maturing yellow fluorescent protein Possible limitations dimer, limited applicability for fusions generation; can form aggregates at high concentration

Recommended filter sets and antibodies

TurboYFP can be recognized using Anti-PhiYFP (Cat.# AB601-AB602) and Anti-PhiYFP(d) (Cat.# AB603-AB604) antibodies available from Evrogen.

TurboYFP can be detected using Omega Optical filter set XF104-3 or Chroma Technology Corp. filter set 42003 ("ZsYellow1").

Performance and use

TurboYFP can be expressed and detected in a wide range of organisms. Mammalian cells transiently transfected with TurboYFP expression vectors give bright fluorescent signals within 8-10 hrs after transfection.

Being overexpressed in long-term culture of cells with high expression levels, TurboYFP shows slight tendency to aggregate. It might limit TurboYFP use in such experimental systems. Please use Phi-Yellow proteins for stable expression and for organelle labeling.

TurboYFP can be expressed and detected in a wide range of organisms. Mammalian cells transiently transfected with TurboYFP expression vectors give bright fluorescent signals within 8-10 hrs after transfection. No cell toxic effects are observed.

TurboYFP can be used in multicolor labeling applications with cyan, green, red, and far-red fluorescent dyes.

		TurboYFP expression in transiently transfected cells: (A) Phoenix cells; (B) Hela cells.

Available variants and fusions

TurboYFP codon usage is optimized for high expression in mammalian cells [Haas et al., 1996], but it can be successfully expressed in many other heterological systems.

Destabilized TurboYFP variant (TurboYFP-dest1): TurboYFP-dest1 is produced by fusing the initial protein with PEST amino acid sequence encoded by region 422-461 of mouse ornithine decarboxylase gene [Li et al., 1998]. This sequence targets the protein to degradation and enables a rapid protein turnover. TurboYFP-dest1 retains spectral properties of the initial protein, but has shorter half-lives (approximately 1.5-2 hrs) as measured by the analysis of fluorescence intensity of cells treated with a protein synthesis inhibitor, cycloheximide. Because of rapid turnover, TurboYFP-dest1 can be used to measure changes in gene expression.

Phi-Yellow proteins: PhiYFP and PhiYFP-m are previous variants of TurboYFP. They possesses same exitation and emission spectra but have a few lesser maturation rate in mammalian cells. Phi-Yellow proteins do not aggregate in long-term cell cultures and in fusions with subcellular localization signals. They have proven suitability to generate stably transfected cell lines.