Yellow fluorescent protein TurboYFP

- Super bright true-yellow fluorescence
- Emission wavelength is ideally positioned between those of green and red fluorescent proteins
- Fast maturation, high pH-stability and photostability
- Destabilized variant is available
- Recommended for gene expression analysis and cell and organelle labeling

TurboYFP is an enhanced variant of the yellow fluorescent protein PhiYFP from jellyfish Phialidium sp. [Shagin et al., 2004]. It possesses superbright yellow fluorescence with emission maximum at 538 nm and is ideally positioned between green and red fluorescent proteins, allowing easy separation of these 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

Mammalian cells transiently transfected with TurboYFP expression vectors give bright fluorescense within 8-10 hrs after transfection. Being overexpressed in long-term culture of cells with high expression levels, TurboYFP shows slight tendency to aggregate. Therefore we recommend that you use parental PhiYFP and PhiYFP-m proteins for long-term expression and organelle labeling.

Despite there dimeric structure, TurboYFP and PhiYFPs perform well in some fusions. Please note, that PhiYFP can be used for generation of fusions to its N-terminus, whereas PhiYFP-m is optimized to generate fusions to its C-terminus. PhiYFP can not be used to generate C-terminal fusions. For protein labeling applications we recommend using specially optimized monomeric TagFPs.

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

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

Available variants and fusions

Variant	Description	Related vector	Cat.#
Humanized TurboYFP	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.	pTurboYFP-C	FP611
		pTurboYFP-N	FP612
		pTurboYFP-B	FP613
		pTurboYFP-PRL	FP615
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-life (approximately 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.	pTurboYFP-PRL-dest1	FP618
		pTurboYFP-dest1	FP619
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.	more