Part:BBa_K5317001
mRuby2
Usage and Biology
The reporter gene mRuby was engineered in 2009 as a variant of the red fluorescent protein eqFP611 by Kredel et al., and developed further to mRuby2 by Lam and colleagues in 2012. Its excitation maxima is at 559 nm and emission maxima is at 600 nm. Due to the large Stokes shift it is well suited for imaging purposes since it reduces spectral overlap when multiple reporters are used as in our case in co-transfection experiments together with EGFP (K3338006). Similar to the findings of Kredel and colleagues (2009) is mRuby2 characterized by a higher resistance to photobleaching compared to the predecessor eqFP611 allowing long-term imaging experiments (Lam et al., 2012).
Our cell-based metal ion sensor employed mRuby2 as a reporter gene fused to the MTF-1 transcription factor (K5317012) to track its cellular localization in the absence and presence of CuSO4. In addition, the fluorescent protein was used to monitor the localization of the transcription factors GraR (K5317020), CcpA (K5317019) and ATF2 (K5317021) in the cells in our antibiotic sensor.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 16
References
Kredel, S., Oswald, F., Nienhaus, K., Deuschle, K., Röcker, C., Wolff, M., Heilker, R., Nienhaus, G. U., & Wiedenmann, J. (2009). mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures. PloS one, 4(2), e4391. https://doi.org/10.1371/journal.pone.0004391
Lam, A. J., St-Pierre, F., Gong, Y., Marshall, J. D., Cranfill, P. J., Baird, M. A., McKeown, M. R., Wiedenmann, J., Davidson, M. W., Schnitzer, M. J., Tsien, R. Y., & Lin, M. Z. (2012). Improving FRET dynamic range with bright green and red fluorescent proteins. Nature methods, 9(10), 1005–1012. https://doi.org/10.1038/nmeth.2171
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