Part:BBa_K1733001
ABI Dimerizable Domain
This part contains ABI (ABA insensitive 1), which is a dimerizable protein domain that will bind with the PYL dimerization domain in the presence of abscisic acid. By fusing these dimerization domains to inert halves of a protein, we were able to control protein function and induce dimerization, and protein activity, with the addition of abscisic acid. We were therefore able to regulate protein activity by taking advantage of these dimerization domains.
The ABI and PYL are found in plants. Thus, it can be tested in mammalian cells, as the ABI-PYL-ABA system is completely orthogonal. We tested this system with our split integrases in mammalian cells.
We used this domain in our experiment to construct split protein complexes. We split the TP901-1 protein and fused the halves to ABI and PYL. We then added abscisic acid into the system to induce dimerization of the domains and the protein halves, and measured the protein activity TP901-1 afterwards. We were able to characterize some split sites as functional, as we regained the TP901-1 activity after inducing dimerization. One functional split site is shown below:
Here, we split the TP901-1 protein between amino acids 326 and 327. We fused the halves to ABI and PYL, and also tested the fusion in reverse orientation. Proper TP901-1 activity would lead to expression of a fluorescent protein. This graph shows that we were able to regain some TP901-1 activity, so we were able to validate this split site as functional.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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