Part:BBa_K4275042
pT7-RBS-OlpB-Ag3-tT7
This composite part is designed for the expression of secondary scaffoldin of the cellulosome complex with the scaffoldin part modified to contain 3 type II cohesin domain and had its anchoring domain changed to an antigen domain. The protein is paired with strong promoter and strong terminator tested suitable for the expression of the coding sequence. The fusion protein is expressed and the antigen domain can interact with nanobody domain to use in cell surface display and the type II cohesin domain can fix the primary scaffoldin subunits to construct cellulosome complexes[1].
Usage and Biology
The protein can also be used in surface display by antigen-nanobody interactions because there is an antigen domain fused at C-terminus. The secondary scaffoldin subunit part contain multiple type II cohesin domain for the specific cohesin-dockerin interactions for fixing the primary scaffold components of the cellulosome complex for assembling of the cellulosome complex nanomachine.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 2058
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 746
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1292
Illegal NgoMIV site found at 1381
Illegal AgeI site found at 223
Illegal AgeI site found at 359
Illegal AgeI site found at 410
Illegal AgeI site found at 542
Illegal AgeI site found at 908
Illegal AgeI site found at 926
Illegal AgeI site found at 1352 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1109
Illegal SapI.rc site found at 1121
Illegal SapI.rc site found at 1745
References
1. Anandharaj, Marimuthu et al. "Constructing A Yeast To Express The Largest Cellulosome Complex On The Cell Surface". Proceedings Of The National Academy Of Sciences, vol 117, no. 5, 2020, pp. 2385-2394. Proceedings Of The National Academy Of Sciences, https://doi.org/10.1073/pnas.1916529117.
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