Part:BBa_K3089013
csgA-linker-mfp5-mfp5-His fusion protein for yeast expression
Csga-linker-mfp5-mfp5-His fusion protein is the purified version of Csga-linker-Mfp5 recombinant protein by His-tag affinity purification (a -Histidine-Histidine-Histidine-Histidine-Histidine-Histidine-Histidine tag fused on the carboxyl terminal). We designed this part by putting together two identical Mfp5 mussel proteins and hoped it shows higher adhesive features than Csga-Mfp5. CsgA is an amyloid-like protein encoded on genome of E.coli MG1655 providing mechanical cohesive strength and Mfp5 is a mussel foot protein from Mytilus galloprovincialis responsible for interface adhesion. This recombinant protein would self-assemble into fibrous bundles or films with adhesive properties by displaying the mussel adhesion domains on the surface of amyloid scaffolds, which would be a promising new generation of bio-inspired adhesives for a wide range of applications. This part was based on the paper "Strong underwater adhesives made by self-assembling multi-protein nanofibres" (Zhong et al, 2014) and has been characterized by iGEM15_TU_Delft (BBa_K1583104). We decided to use another chassis for the same protein rather than solely using E.coli BL21(DE3) because of relatively low and harder protein expression.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 274
Illegal PstI site found at 23 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 274
Illegal PstI site found at 23 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 274
Illegal XhoI site found at 244 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 274
Illegal PstI site found at 23 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 274
Illegal PstI site found at 23 - 1000COMPATIBLE WITH RFC[1000]
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