Part:BBa_K1127000
A3-Flg Gold binding peptide
A de novo synthesized gene with A3 and Flg domains which should hypothetically bind to gold and form nanoparticles.
Characterization
We created four variants of the fusion between MIDAS-2 and A3. We also include the N-terminal pelB tag for secretion to the periplasm and IM9 for higher stability. Note that the tag should be removed during transportation process and potentially our peptides should be released to the extracellular space. IM9 tag is relatively small (~9.5 kDa) and should not interfere with mineralization mediated by the peptides.
For peptide activity testing , we cloned the peptides under the constitutive promoter (BBa_J23100). Characterization using Nanoparticle Tracking Analyzer (NanoSight LM10 ®). The device allows us to visualize, measure and characterize any type of nanoparticles and any size down to 10nm. The technology is based on optical properties as well as Brownian motion of the particles. Please visit http://www.nanosight.com/ for more information.
We obtained a series of recordings and several analytical reports of the particles within samples from the overnight cultures of E. coli strain DH5 alpha transformed with the hybrids. Unfortunately, the results can't provide evidence for gold bio-mineralization because changes were subtle and not significant compared to the controls (untransformed E. coli). Please visit out Protocols section for more detail [http://2013.igem.org/Team:York_UK/Notebook.html?page=protocols].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 78
- 1000COMPATIBLE WITH RFC[1000]
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