Yellow fluorescent protein Dyes
Yellow Fluorescent Protein (YFP) can be regarded as a mutant of green fluorescent protein, which originally originated from Aequorea Victoria, compared to green fluorescent protein, its fluorescence shifts to the red spectrum, and this is mainly due to the change of threonine at position 203 of the protein to tyrosine. Its maximum excitation wavelength is 514 nm and its maximum emission wavelength is 527 nm.
Figure 1. Protein structure of Yellow Fluorescent Protein (YFP).
History
The study of the yellow fluorescent protein family began after the threonine at position 203 was found to be close to the chromophore in the crystal structure of green fluorescent protein. Mutation of threonine at position 203 to tryptophan can stabilize the dipole moment of the chromophore in the excited state, which can increase the wavelength of both the excitation light and the emitted light by 20 nm. And further transformed into enhanced yellow fluorescent protein (EYFP). EYFP is one of the most widely used and brightest fluorescent proteins. The organic combination of the brightness and emission spectrum of EYFP makes this fluorescent probe well used in fluorescent multicolor imaging. EYFP can also be applied to FRET with enhanced cyan fluorescent protein (ECFP) or GFP2. However, yellow fluorescent protein also has some problems. For example, it is very sensitive to acidic conditions and loses nearly 50% of its fluorescence at pH 6.5. In addition, EYFP is very sensitive to chloride ions, and photobleaching is more severe than green fluorescent protein.
Introductions
Yellow fluorescent protein (YFP) is a genetic mutant of green fluorescent protein (GFP), originally from Aequorea victoria. Its excitation peak is 514 nm and its emission peak is 527 nm. Like the parent GFP, YFP is a useful tool in cell and molecular biology due to its properties that can be used for fluorescence microscopy. The three improved versions of YFP are Citrine, Venus, and Ypet. They have reduced chloride sensitivity, faster maturity, and higher brightness (defined as the product of extinction coefficient and quantum yield). Generally, YFP is used as a receptor for a gene-encoded FRET sensor, and its most likely donor FP is monomeric cyanofluorescent protein (mCFP). The redshift relative to GFP is caused by the Pi-Pi stacking interaction, which is the result of the substitution of T203Y introduced by the mutation, which substantially increases the polarization of the local chromophore environment and provides Added extra electron density. "Venus" contains a new amino acid substituent -F46L, which accelerates the oxidation of the chromophore at 37 ° C, a mature rate-limiting step. The protein has other substitutions (F64L/M153T/V163A/S175G) that makes Venus fold well and makes it relatively resistant to acidosis and Cl-.
Applications
Like green fluorescent protein, YFP is a very commonly used reporter gene in cell biology and molecular biology. Currently, there are three improved yellow fluorescent proteins: Citrine, Venus, and Ypet. These three improved proteins are brighter, more stable, and mature faster, so they are widely used. Yellow fluorescent protein is most commonly used for fluorescence resonance energy transfer as an acceptor of fluorescent energy.
Reference:
- Nagai, T.; et al. A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications. Nature Biotechnology. 2002, 20 (1): 87–90.
Dyes Application
- Acridine Dyes
- Alcian Blue Stain Dyes
- Alexa Fluor Dyes
- Allophycocyanin Dyes
- Anthracene Dyes
- BODIPY Dyes
- Coumarin Dyes
- Cresyl Violet Dyes
- Crystal Violet Dyes
- DAPI Dyes
- DNA Dyes Stain
- Feulgen Stain Dyes
- Fluorescein Isothiocyanate Dyes
- Fluorescent protein Dyes
- Nile Blue Dyes
- Nile Red Dyes
- Pacific Blue Dyes
- Pacific Green Dyes
- Pacific Orange Dyes
- Pyrene Dyes
- Texas Red Dyes
- Xanthene Dyes
Probe Application
