Team:Paris Bettencourt/Proof
Proof-of-concept - developing fabric-specific bindig domains (Fabric Binding Domains)
As our proof-of-concept we show that the Fabric Binding Domains (FBD) that we developed in this project work, by having affinity and specificity for a given type of fabric.
The complete list of FBDs follows. A more broader description of each FBD can be found in their Biobricks' page.
| Enzymes for Stain Digestion | |||
|---|---|---|---|
| BioBrick ID | Enzyme name | Description | Reference |
| BBa_K2043011 | GFP-FBD2 | GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD2 | |
| BBa_K2043012 | GFP-FBD5 | GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD5 | |
| BBa_K2043013 | GFP-FBD6 | GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD6 | |
| BBa_K2043014 | GFP-FBD7 | GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD7 | |
| BBa_K2043016 | GFP-FBD9 | GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD9 | |
| BBa_K2043017 | GFP-FBD10 | GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD10 | |
The following figure is proof that the FBDs work. Our results show that each FBD has an individual relationship with each of the fabrics, some having affinity to many, other showing specificity to a given fabric.
Figure 5: Analysis of FBD-GFP fusion proteins. With this image we show that the GFP activity was not affected by the fusion between the FBDs and the GFP. CBD stands for the +control, cellulose binding domain from the Biobrick registry BBa_K1321357.
Cell extract from strains expressing our fusion proteins was incubated overnight with the fabrics. Afterwards two washes of the fabrics were performed. Water, PBS, BSA 5%, Ethanol 70% and Catechol correspond to the washing solutions we used to remove the non-bound GFP, as well as to test the binding strength of the different peptides. The data displayed corresponds to the values after the final wash, normalised using the values from the first wash.
The intensity of the colour corresponds to the GFP signal measured at that point.
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