Difference between revisions of "Part:BBa K1745001:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | KaiB and KaiC are in reverse direction of KaiA to maximize the efficiency of the double terminator Part:BBa_B0014 | + | KaiB and KaiC are in reverse direction of KaiA to maximize the efficiency of the double terminator Part:BBa_B0014. In vitro analysis of the Kai system found that the robustness of the oscillator was sensitive to KaiA:KaiC stoichiometry. In order to investigate this ratio, KaiA expression is driven under an L-rhamnose inducible promoter Part BBa_K914003 while KaiB and KaiA are driven under a constitutive promoter Part BBa_J23100. |
===Source=== | ===Source=== |
Revision as of 00:37, 18 September 2015
KaiABC Oscillator
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 3014
Illegal NheI site found at 3037 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 410
Illegal XhoI site found at 477 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2945
- 1000INCOMPATIBLE 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. In vitro analysis of the Kai system found that the robustness of the oscillator was sensitive to KaiA:KaiC stoichiometry. In order to investigate this ratio, KaiA expression is driven under an L-rhamnose inducible promoter Part BBa_K914003 while KaiB and KaiA are driven under a constitutive promoter Part BBa_J23100.
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