Part:BBa_K1075014
RBS32-VinD1-sspB[XB]-TT
This part contains the D1 domain of Vincolin fused with the XB part of the split version of sspB.
Usage and Biology
The D1 domain of mammalian Vincolin forms a tight dimer with the ipaA peptide.
EcsspB itself regulates the degradation of ssrA tagged proteins through the ClpXP protease in procaryotes. In engineered systems it is used to induce degradation of specifically ssrA tagged proteins. The XB domain of sspB is part of the split system of E. Coli sspB (EcsspB) which was used in fusion with FKBP and FRB to induce the activity of sspB and therefore degradation of proteins with Rapamycins. [1] EcsspB can be functionally divided in three parts: -the N terminal Core domain (113 AA) -the C terminal XB peptide (25 AA) -and a 'flexible linker' in between the first parts ( We used the same domain structure as is used within the Rapamycin inducable split system. While the Core domain is responsible for dimerization of sspB and binding of ssrA the XB domain binds the protease ClpXP. The flexible linker is called flexible because it was found that an increase or reduction in size or amino acid composition does not influence the function of sspB as much as it would in the other domains.
The part can be used to regulate the dimerization of the two parts of the split sspB. As Vincolin bins to ipaA, the sspB dimer is stabilized.
We used this part to achieve a light-induced dimerization of the sspB parts. Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 75
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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