Measurement

Part:BBa_K1051288

Designed by: Rui Guan   Group: iGEM13_Shenzhen_BGIC_ATCG   (2013-09-01)
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The measurement pathway of ubiquitin fusion protein.

The measurement pathway of ubiquitin fusion protein.

http://upload.wikimedia.org/wikipedia/commons/thumb/5/5b/Ubiquitylation.svg/800px-Ubiquitylation.svg.png
The ubiquitination system.
Ubiquitin,a small regulatory protein that has been found in almost all tissues of eukaryotic organisms,Ubiquitination is a post translational modification where ubuiquitin is attached to a substrate protein.It is carried out in three main steps; activation, conjugation and ligation, performed by ubiquitin-activating enzymes (E1s), ubiquitin-conjugating enzymes (E2s) and ubiquitin ligases (E3s).
We want to install ubiquitin to fluorescent proteins at N-terminal.
To test the function of ubiquitin fusion protein, we construct this measurement pathway K1051288, which contains J63006, K1051200,K1051003 and K1051006. Growth-Curve.jpg
Yeast growth curve. Sc_bright_20.jpg
We made the chip as a platform for watching and synchronize the cells. First step is capture the cells by the chip. As showed in the figure, budding yeast can be captured by the chip successfully.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 788
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 150
    Illegal AgeI site found at 580
    Illegal AgeI site found at 808
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2351


References

[1] Wilkinson KD (October 2005). "The discovery of ubiquitin-dependent proteolysis". Proc. Natl. Acad. Sci. U.S.A. 102 (43): 15280–2. doi:10.1073/pnas.0504842102. PMC 1266097. PMID 16230621.
[2] Kimura Y, Tanaka K (2010). "Regulatory mechanisms involved in the control of ubiquitin homeostasis". J Biochem 147 (6): 793–8. doi:10.1093/jb/mvq044. PMID 20418328.

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Parameters
n/aThe measurement pathway of ubiquitin fusion protein.