Part:BBa_K3956006
scl2 enzyme cleaving domain (thrombin/trypsin)
Encodes LVPRGSP, used to separate the non-collagenous (V) domain of scl2 collagen from the desired triple-helical collagen-like domain (CL).
Usage and Biology
This part was used in CCA_San_Diego’s 2021 CollaGene project to cleave the globular domain of synthetic bacterial collagen from the collagen-like domain. This process is crucial to separate the functional triple helices from the stabilizing N-terminus. This also mimics a natural MMP collagenase cut site in human collagen. It was added in between a globular V domain and a collagen-like CL domain documented by Peng et al. in 2012. Although we were able to verify that the construct had been correctly inserted into the plates by colony PCR verification, the protein expression was not able to be verified within the time constraints of the competition.
It is interesting to note, however, that when mammalian collagen was cleaved with pepsin (which is also an enzyme that can be used to cleave this construct), a similar length protein was visualized in an SDS-PAGE, indicating that the CL domains of bacterial and mammalian collagen are very similar.
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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