Part:BBa_K2635003
GBP C-terminal
Green fluorescent protein (GFP) and GFP-binding protein (GBP) system is a very common tool in synthetic biology. We use GFP-nanobody(GBP), a single-chain VHH antibody domain developed with specific binding activity against GFP, because it is the most well-studied nanobody. In our design, we segregate GFP-nanobody into two fragments, C-terminal and N-terminal outside the membrane. Both fragment have it own C-terminal or N-terminal TEV protease fragment linked by TM domain. When GFP appears, both of the C-terminal and N-terminal of GBP bind to it. This movement facilitates the combination of C-terminal and N-terminal TEV protease in the cytosol, and then the intact TEV protease starts to cut at TEV cleavage site. [1][2]
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 BsaI site found at 53
References
- ↑ Kubala, M. H., Kovtun, O., Alexandrov, K., &Collins, B. M. (2010). Structural and thermodynamic analysis of the GFP : GFP-nanobody complex, 19(September), 2389–2401. https://doi.org/10.1002/pro.519
- ↑ Tang, J. C. Y., Rudolph, S., Dhande, O. S., Abraira, V. E., Choi, S., Lapan, S. W., …Cepko, C. L. (2015). Cell type-specific manipulation with GFP-dependent Cre recombinase. Nature Neuroscience, 18(9), 1334–1341. https://doi.org/10.1038/nn.4081
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