Our parts were designed for the two main branches of our labwork - in vivo expression and export to try and achieve structural colour, and overexpression for purification and in vitro studies of our recombinant reflectins.
Overexpression for purification
Reflectins are a family of proteins which help give cephalopods their amazing camouflage abilities and which can self-assemble into a variety of multilayered structures with optical properties. We worked with Reflectin A1 from Loligo pealeii. See our background page for more information about reflectins and their role in cephalopod camouflage.
In order to isolate pure reflectin, we built and submitted Poly-His tagged Reflectin A1 generator. In pure form reflectins may be used to create vibrantly coloured thin films and other devices. We used a poly-His affinity column to purify reflectin from cells for this purpose.
This BioBrick was created in order for us to control for the expression of reflectin and its location in the cell. Using it, we were able to determine that reflectin forms inclusion bodies when expressed at a high level.
We submitted a variant of the TorA leader sequence export tag. This TorA leader sequence variant has had been successfully used to export GFP to the periplasm of E.coli as described here. Our tag differs slightly from the TorA tag already in the registry and should be cheaper to obtain from primer synthesis.
We also submitted Reflectin A1 with N-terminal TorA tag and Reflectin A1 with N-terminal TorA tag and C-terminal sfGFP, which we used in our periplasm export attempt.
...and the rest
A complete list of parts submitted to the Registry by this year's Cambridge team can be found below. (We also added our experience of part I0500 to the registry.)