Difference between revisions of "Part:BBa K1745001:Design"

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===Design Notes===
 
===Design Notes===
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In vitro investigations of this modular system revealed that the robustness of oscillations was sensitive to stoichiometric ratios of KaiA to KaiC (Nakajima, Science 2015). To experiment with these stoichiometric ratios, KaiB and KaiC expression are driven under a constitutive promoter (Part J23100) while KaiA expression is driven under a L-rhamnose inducible promoter (Part K914003). 
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KaiB and KaiC are in reverse direction of KaiA to maximize the efficiency of the double terminator Part:BBa_B0014
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===Source===
 
===Source===

Revision as of 00:30, 18 September 2015


KaiABC Oscillator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3014
    Illegal NheI site found at 3037
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 410
    Illegal XhoI site found at 477
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2945
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 540


Design Notes

KaiB and KaiC are in reverse direction of KaiA to maximize the efficiency of the double terminator Part:BBa_B0014

Source

The KaiABC circadian oscillator is endogenous to the cynanobacterial species Synechococcus elongatus.

References

Chen, A. H., Lubkowicz, D., Yeong, V., Chang, R. L., & Silver, P. a. (2015a). Transplantability of a circadian clock to a noncircadian organism. Science Advance, 1(5), 1–6. http://doi.org/10.1126/sciadv.1500358

Nakajima, M., Imai, K., Ito, H., Nishiwaki, T., Murayama, Y., Iwasaki, H., … Kondo, T. (2005). Reconstitution of circadian oscillation of cyanobacterial KaiC phosphorylation in vitro. Science (New York, N.Y.), 308(5720), 414–415. http://doi.org/10.1126/science.1108451

Nakajima, M., Ito, H., & Kondo, T. (2010). In vitro regulation of circadian phosphorylation rhythm of cyanobacterial clock protein KaiC by KaiA and KaiB. FEBS Letters, 584(5), 898–902. http://doi.org/10.1016/j.febslet.2010.01.016

Pattanayak, G., & Rust, M. J. (2014). The cyanobacterial clock and metabolism. Current Opinion in Microbiology, 18(1), 90–95. http://doi.org/10.1016/j.mib.2014.02.010

Rust, M. J., Markson, J. S., Lane, W. S., Fisher, D. S., & O’Shea, E. K. (2007). Ordered phosphorylation governs oscillation of a three-protein circadian clock. Science (New York, N.Y.), 318(5851), 809–812. http://doi.org/10.1126/science.1148596

Taniguchi, Y., Takai, N., Katayama, M., Kondo, T., & Oyama, T. (2010). Three major output pathways from the KaiABC-based oscillator cooperate to generate robust circadian kaiBC expression in cyanobacteria. Proceedings of the National Academy of Sciences of the United States of America, 107(7), 3263–3268. http://doi.org/10.1073/pnas.0909924107