Difference between revisions of "Part:BBa K5317001"
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===Usage and Biology===
 
===Usage and Biology===
  
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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 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 ((<span class="plainlinks">[https://parts.igem.org/Part:BBa_K3338006 K3338006]</span>)).  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).  
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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 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 (<span class="plainlinks">[https://parts.igem.org/Part:BBa_K3338006 K3338006]</span>).  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).
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Our cell-based metal ion-sensor employed mRuby2 as a reporter gene fused to the MTF-1 transcription factor (<span class="plainlinks">[https://parts.igem.org/Part:BBa_K5317012 K5317012]</span>) to track its cellular localization without and in the presence of CuSO4.  
  
 
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Revision as of 09:44, 16 September 2024


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 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 without and in the presence of CuSO4.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE 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