Team:Cambridge/Parts

From 2011.igem.org

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==System diagrams==
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Our parts were designed for the two main branches of our labwork - [[Team:Cambridge/Project/In_Vivo | ''in vivo'' expression and export]] to try and achieve structural colour, and [[Team:Cambridge/Project/In_Vitro | overexpression for purification]] and ''in vitro'' studies of our recombinant reflectins.
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===Overexpression for purification===
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[[File:Cam_Overexpression_Construct.png | center | 600px]]
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===Export===
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|You can write a background of your team here.  Give us a background of your team, the members, etc.  Or tell us more about something of your choosing.
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[[File:Cam_Export_Construct.png | center | 600px]]
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|[[Image:CAM_Bactiridescence_logo_Small.png|200px|right|frame]]
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''Tell us more about your project.  Give us background.  Use this is the abstract of your project.  Be descriptive but concise (1-2 paragraphs)''
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|[[Image:Team_photo.png|x200px|right|frame|Your team picture]]
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|align="center"|[[Team:Cambridge | Team Example]]
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==Featured Parts==
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===Parts===
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[[Image:Cam_Multilayer_drop_1.jpg|400px|right|thumb| A thin film composed of Reflectin A1 from Loligo pealeii, purified using a poly-His affinity column]]
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New for iGEM 2010 is the ''groupparts'' tag.  This tag will generate a table with all of the parts that your team adds to your team sandbox.  Note that if you want to document a part you need to document it on the [http://partsregistry.org Registry], not on your team wiki.
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===[http://partsregistry.org/wiki/index.php?title=Part:BBa_K638001 Reflectin A1]===
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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 [[Team:Cambridge/Project/Background | background page]] for more information about reflectins and their role in cephalopod camouflage.
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In order to isolate pure reflectin, we built and submitted [http://partsregistry.org/wiki/index.php?title=Part:BBa_K638202 Poly-His tagged Reflectin A1 generator]. In pure form reflectins may be used to create [[Team:Cambridge/Project/In_Vitro | vibrantly coloured thin films]] and other devices. We used a poly-His affinity column to purify reflectin from cells for this purpose.
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<br style='clear:both' />
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[[File:Cam Reflectin-GFP-inclusionbodies.jpg | thumb | 175px | left | ''E. coli'' transformed with our pBAD-ReflectinA1-GFP construct]]
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===[http://partsregistry.org/wiki/index.php?title=Part:BBa_K638301 Reflectin A1 GFP fusion]===
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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 [[Team:Cambridge/Project/In_Vitro#Over-Expression | inclusion bodies]] when expressed at a high level.
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===[http://partsregistry.org/wiki/index.php?title=Part:BBa_K638402 Improved TorA tag]===
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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 [http://www.ncbi.nlm.nih.gov/pubmed/11123687 here]. Our tag differs slightly from [http://partsregistry.org/wiki/index.php?title=Part:BBa_K233307 the TorA tag already in the registry] and should be cheaper to obtain from primer synthesis.
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We also submitted [http://partsregistry.org/wiki/index.php?title=Part:BBa_K638401 Reflectin A1 with N-terminal TorA tag] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K638403 Reflectin A1 with N-terminal TorA tag and C-terminal sfGFP], which we used in our [[Team:Cambridge/Project/In_Vivo#Periplasmic_Export | periplasm export]] attempt.
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===...and the rest===
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A complete list of parts submitted to the Registry by this year's Cambridge team can be found below.
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(We also added our [http://partsregistry.org/Part:BBa_I0500:Experience experience of part I0500] to the registry.)
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<link rel="stylesheet" type="text/css" href="/common/tablesorter/themes/groupparts/style.css" />
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</html>
<groupparts>iGEM011 Cambridge</groupparts>
<groupparts>iGEM011 Cambridge</groupparts>
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Latest revision as of 00:16, 22 September 2011

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OVERVIEW
home

Contents

System diagrams

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

Cam Overexpression Construct.png

Export

Cam Export Construct.png

Featured Parts

A thin film composed of Reflectin A1 from Loligo pealeii, purified using a poly-His affinity column

Reflectin A1

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.

E. coli transformed with our pBAD-ReflectinA1-GFP construct

Reflectin A1 GFP fusion

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.

Improved TorA tag

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.)

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