Difference between revisions of "Part:BBa K2043012"
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https://static.igem.org/mediawiki/parts/2/20/Paris_Bettencourt-biobricks_gfp5.png<br><br> | https://static.igem.org/mediawiki/parts/2/20/Paris_Bettencourt-biobricks_gfp5.png<br><br> | ||
| − | FBD-5 is highly specific to all fabrics in most conditions. This amino acid sequence has a positive charge, and a Hydropathicity (GRAVY) index of -0.614 (slightly hydrophilic).<br> | + | FBD-5 has a sequence of ADIRHIK, is highly specific to all fabrics in most conditions. This amino acid sequence has a positive charge, and a Hydropathicity (GRAVY) index of -0.614 (slightly hydrophilic).<br> |
These experiments were performed by obtaining cell extract of cells of a strain of <i>E. coli<(i> overexpressing FBD5-GFP, incubating the cell extract with the fabrics overnight, and then performing 2 washes with the indicated solutions. | These experiments were performed by obtaining cell extract of cells of a strain of <i>E. coli<(i> overexpressing FBD5-GFP, incubating the cell extract with the fabrics overnight, and then performing 2 washes with the indicated solutions. | ||
Revision as of 16:10, 25 October 2016
GFP-FBD5
In the context of Paris Bettencourt 2016 iGEM project, Frank&Stain, we looked for fabric binding domains with phage display in order to optimise our enzymes on the topic of pigment degradation. More information can be consulted in the wiki, but we wanted to degrade wine stains, and we developed the FBD in order to direct the enzymatic activity to the fabric.
We fused the GFP gene with the FBD5 (in the N-terminal of the GFP) in order to understand two things: (i)if the GFP fluorescence would be affect by the binding of the FBD and (ii)if the binding capacity would be affected.
The results for FBD5-GFP are as follows:
FBD-5 has a sequence of ADIRHIK, is highly specific to all fabrics in most conditions. This amino acid sequence has a positive charge, and a Hydropathicity (GRAVY) index of -0.614 (slightly hydrophilic).
These experiments were performed by obtaining cell extract of cells of a strain of E. coli<(i> overexpressing FBD5-GFP, incubating the cell extract with the fabrics overnight, and then performing 2 washes with the indicated solutions.
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]
GFP-FBD5
Jain, P., Soshee, A., Narayanan, S. S., Sharma, J., Girard, C., Dujardin, E., & Nizak, C. (2014). Selection of arginine-rich anti-gold antibodies engineered for plasmonic colloid self-assembly. The Journal of Physical Chemistry C, 118(26), 14502-14510.
Soshee, A., Zürcher, S., Spencer, N. D., Halperin, A., & Nizak, C. (2013). General in vitro method to analyze the interactions of synthetic polymers with human antibody repertoires. Biomacromolecules, 15(1), 113-121.
Boyer, S., Biswas, D., Soshee, A. K., Scaramozzino, N., Nizak, C., & Rivoire, O. (2016). Hierarchy and extremes in selections from pools of randomized proteins. Proceedings of the National Academy of Sciences, 201517813.