Team:Cambridge/Project

From 2011.igem.org

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Bact<b>iridescence</b> was based around the properties of [[Team:Cambridge/Project/Background | reflectin]], a squid protein with the highest refractive index of any known proteinaceous substance. In squid this protein forms complex platelets which act as Bragg reflectors to provide camouflage.
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Bact<b>iridescence</b> was based around the properties of [[Team:Cambridge/Project/Background | reflectin]], a squid protein with the highest refractive index of any known proteinaceous substance. In squid this protein forms complex platelets which act as [http://en.wikipedia.org/wiki/Bragg_reflector Bragg reflectors] to provide camouflage.
===Project Goals===
===Project Goals===
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===[[Team:Cambridge/Project/In_Vitro | In Vitro]]===
===[[Team:Cambridge/Project/In_Vitro | In Vitro]]===
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[[File:Cam_Multilayer_drop_1.jpg | right | thumb | 150px | A multilayer thin film]]
By engineering ''E. coli'' to overexpress reflectins we have;
By engineering ''E. coli'' to overexpress reflectins we have;
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*Purified reflectin and documented best practice for high purity yields.
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*[[Team:Cambridge/Experiments/Protein_Purification | Purified reflectin]] and documented best practice for high purity yields.
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*Made thin films which show structural colours.
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*Made [[Team:Cambridge/Project/Microscopy#Reflectin_Thin_Films | thin films]] which show structural colours.
*Demonstrated the rapid colour changes possible with reflectin.
*Demonstrated the rapid colour changes possible with reflectin.
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**Videos of our thin films are available on [http://www.youtube.com/user/cambridgeigem2011 youtube].
===[[Team:Cambridge/Project/Gibthon | Software]]===
===[[Team:Cambridge/Project/Gibthon | Software]]===
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[[File:Gibthon2.0beta.png | left | thumb | 100px | The Gibthon logo]]
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We contributed to Gibthon to help create an intuitive set of tools for designing constructs, fully compatible with both BioBrick standards and newer assembly techniques.
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We contributed to [http://www.gibthon.org/ Gibthon], an open-source collection of web-based tools for construct design, fully compatible with both BioBrick standards and newer assembly techniques.
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*Greatly improved import and display of fragments.
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*Greatly improved import and display of fragments (including support for [http://partsregistry.org/Main_Page partsregistry.org]).
*Added tools to allow management of uploaded parts.
*Added tools to allow management of uploaded parts.
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<html><div style='clear:both'></div></html>
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===[[Team:Cambridge/Project/Microscopy | Microscopy]]===
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==[[Team:Cambridge/Project/Future | Future work]]==
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Initially we looked at some squid tissue using a confocal microscope, to see the optical effect of reflectins in squid cells.  We also imaged bacterial cells engineered to produce reflectins.
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===[[Team:Cambridge/Project/In_Vitro | In Vitro]]===
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When we over-expressed reflectin in E.Coli, we found (by making a GFP fusion) that while reflectin is surprisingly non-toxic to E.Coli, it formed inclusion bodies. We then extracted these inclusion bodies, purified the protein using a number of different techniques, and made [[Team:Cambridge/Experiments/Reflectin_Thin_Films_I | thin films]] by spin-coating and flow-coating.
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===[[Team:Cambridge/Project/In_Vivo | In Vivo]]===
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In order to express reflectin at lower levels, we made an arabinose-inducible  version, both with and without a GFP-fusion. We found that at lower levels of expression, the reflectin-GFP fusion would not form inclusion bodies, but appeared to be uniformly distributed throughout the cell [https://2011.igem.org/Team:Cambridge/Project/Microscopy (see the Microscopy page)] . We made several attempts to image this – hoping to find some change in optical properties – but found that while the induced cells did appear to exhibit iridescence, so did the uninduced. This, we theorise, is due to thin film interference around the cell wall and membrane.
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We also attempted to export both our reflectin and our reflectin-GFP to the periplasm, in the hope that this environment would be more similar to the environment in which reflectin naturally folds and that the small space will promote reflectin's membrane-associating properties. As of writing, this has yet to be successful.
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===[[Team:Cambridge/Project/Conclusion | Conclusion and Future Work]]===
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By creating the first BioBrick parts for production of structural colour, we hope to facilitate further research. Although time did not allow us to explore the full potential of our project, we have some ideas for what could be done next.
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Our project attempts to lay some groundwork for future research in to reflectins. Reflectin has numerous possible future applications, from display technologies to rapid bio-reporters.  
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Latest revision as of 02:52, 22 September 2011

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OVERVIEW
home
Bactiridescence was based around the properties of reflectin, a squid protein with the highest refractive index of any known proteinaceous substance. In squid this protein forms complex platelets which act as [http://en.wikipedia.org/wiki/Bragg_reflector Bragg reflectors] to provide camouflage.

Contents

Project Goals

We aimed to express reflectin in E. coli and to investigate its optical properties in order to build the groundwork for the manipulation of living structural colour. We also looked at the over-expression of reflectin in E. coli, in order to obtain relatively pure samples of the protein for making thin films.

Much of our work (particularly the in vivo work) simply hadn't been tried before, so, while we had high hopes, we could not be sure as to what would happen.

Achievements

In one short summer the 2011 Cambridge team has produced a set of BioBrick parts to allow future researchers to explore synthetic biology applications for structural colour.

In Vivo

Working with living cells we have;

In Vitro

A multilayer thin film

By engineering E. coli to overexpress reflectins we have;

  • Purified reflectin and documented best practice for high purity yields.
  • Made thin films which show structural colours.
  • Demonstrated the rapid colour changes possible with reflectin.
    • Videos of our thin films are available on [http://www.youtube.com/user/cambridgeigem2011 youtube].

Software

The Gibthon logo

We contributed to [http://www.gibthon.org/ Gibthon], an open-source collection of web-based tools for construct design, fully compatible with both BioBrick standards and newer assembly techniques.

  • Greatly improved import and display of fragments (including support for [http://partsregistry.org/Main_Page partsregistry.org]).
  • Added tools to allow management of uploaded parts.

Future work

By creating the first BioBrick parts for production of structural colour, we hope to facilitate further research. Although time did not allow us to explore the full potential of our project, we have some ideas for what could be done next.


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