mKate is a fluorophor derived from dsRed
mKate is a fluorophor derived from dsRed and is a FRET partner of LssmOrange [BBa_K1055001].
ATTENTION: If you want to make a fusion protein with this part, please refer to our experiences!
Usage and Biology
mKate is a red fluorescent protein that we want to use as a acceptor for FRET (Figure 3). According to Evrogen mKate has an excitation maximum at 588 nm and an emission maximum at 633 nm. Figure 2 shows that the excitation maximum is at 579 nm and the emission maximum is 611 nm. The emission maximum is big enough to get stimulated by LSSmOrange, the other fluorescent protein. Of course, we checked also the emission and excitation of our BL21(DE3) cells (data not shown) and we can eliminate the theories that these cells disturb this florescence measurement.
Moreover, unlike for our LssmOrange BioBrick (BBa_K1055001), there is a crystal structure available for further analysis in the [RCSB database] which is shown below:
The expression of mKate is not toxic towards the bacterial host E.coli Bl21(DE3). The growth rate was in a usual time scale of approximately 30 minutes.
Daria M. Shcherbakova et al. (2012) An Orange Fluorescent Protein with a Large Stokes Shift for Single-Excitation Multicolor FCCS and FRET Imaging. J. Am. Chem. Soc. 134 (18), 7913–7923
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
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- 1000COMPATIBLE WITH RFC