Difference between revisions of "Part:BBa K1976002"
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Revision as of 22:12, 21 October 2016
mVenus with a LVA degradation tag
This part contains an E. coli optimized coding sequence of the yellow fluorescent protein mVenus with an associated LVA degradation tag.Usage
In some cases a fast signaling reporter, as well as a fast decay of the said reporter is necessary to create a specific genetic circuit. One of the best fitting reporters might be the E.coli optimized version of mVenus. With an average maturation time of 40 min in vitro it is faster than GFP. In order to prevent a persistent fluorescence after the expression of the reporter stopped, mVenus is expressed with a LVA degradation tag to decrease the protein half-life. Another positive aspect of mVenus is the lowered sensitivity towards pH and chloride ion concentration, one of the drawbacks of wild-type GFP. The lack of disulfide bonds enables fluorescence under reductive conditions. Moreover, the reporter is not regulated by any proteins, cofactors or substrates. Therefore mVenus does not only expand the spectrum of fluorescence proteins in the registry, it also is a good alternative for various genetic circuits.
Characteristica
Below are some characteristica of mVenus listed.
Figure 1: Excitation and emission spectrum of mVenus.The fluorescent protein excitation intensity at 280 nm is thought to be due to fluorescence resonance energy transfer from aromatic amino acids to the chromophore.
As seen in Figure2,the maximum of exication is at 515 nm and the maximum of emission at 528 nm. mVenus has a brightness of 15, which is defined as the roduct of the extinction coefficient and the quantum yield at pH 7.4 under ideal maturation conditions. Its photostabillity is 15 s, where the photostabillity is defined as the time for bleaching from an initial emission rate of 1,000 photons/s down to 500 photons/s. its pka is located at 6.0. it is a weak dimer.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
1.
2. Pabak Sarkar, Srinagesh V. Koushik et al. (2009) Photophysical Properties of Cerulean and Venus Fluorescent Proteins - J Biomed Opt. 2009 ; 14(3): 034047. doi:10.1117/1.3156842
3. Nathan C Shaner1, Paul A Steinbach & Roger Y Tsien (2005) A guide to choosing fluorescent proteins - Nature Methods