Part:BBa_K2155009
RFDock
Dockerin
Usage and Biology
It is from from the R. flavefaciens ScaA (787−879 amino acids, GenBank Accession number: CAC34384.3), and can automatically assemble with a cohesin module from ScaB. In our project, we use Synthetic Protein Scaffolds to improve the efficiency of enzyme catalytic reaction. This dockerin module is a part of Synthetic Protein Scaffolds. It can automatically assemble with a matching cohesin module of Synthetic Protein Scafflods.
Contribution From UESTC-China 2021
Group: iGEM Team UESTC-China 2021
Author: Liyan Wu
Characterization from UESTC-China 2021
We use this dockerin domain from C.thermocellum to connected with xylanase by a (GGGGS)3 linker, and it can bind with CtCohesin which is part of our scaffold protein, and ultimately anchor our cellulase and xylanase to the cell surface of Pichia pastoris to improve efficiency of deink. Because we chose Pichia pastoris as the chassis organism, we have done codon optimized for yeast expression, and our part number is BBa_K3819015.
- Figure 1. Gene squence alignment of CtDockerin from iGEM15_Concordia and UESTC-China
- Figure 2. Amino acid squence alignment of CtDockerin from iGEM15_Concordia and UESTC-China
The two figures above show that the amino acid sequence of the two parts are the same but the gene sequence is not completely the same.
We finally successfully express the CtDockerin domain, as a 6×His tag has been linked, we can observe its expression as the following results of Dot-Blot analysis.
- Figure 3. The Dot-Blot results of recombined GS115(pXynB) fermentation broth. The sample was gained after fermentation in YPD for 96h. XynB(His-Tag) was tested by adding 2ul fermentation supernatant on NC membrane and exposing 60s.
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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