Team:Cambridge/Experiments/Initial Exercise Group One

From 2011.igem.org

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(Primer Design)
(Initial Exercise: Katy, Heather, Marta and Veronica)
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[[File:Cam_Oligo design_sketch.jpeg|frameless|700px|A sketch of design concept]]
[[File:Cam_Oligo design_sketch.jpeg|frameless|700px|A sketch of design concept]]
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===Primer Design===
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used the Gibthon Construct Designer developed by the 2010 iGEM Cambridge team
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[[File:name.fmt|frameless|700px|Left side of Div IVA sequence]]
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{{Template:Team:Cambridge/CAM_2011_TEMPLATE_FOOT}}
{{Template:Team:Cambridge/CAM_2011_TEMPLATE_FOOT}}

Revision as of 23:02, 6 July 2011

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OVERVIEW
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Contents

Initial Exercise: Katy, Heather, Marta and Veronica

Fusion of Div IVA protein with GFP

Role of Div IVA

Div IVA controls septum positioning in Bacillus Subtilis

Mutants show

  • inhibition of cell division initiation*
  • abnormally placed septum*
  • some small anucleate cells*

Reasons for Choice

  • constitutively expressed*
  • Distinct characteristic distribution that is easy to observe*
  • successful divIVA-GFP fusion documented by Edwards and ERrington, 1997*

Construct Design

We wish to:

  • keep the original promotor for the Div IVA*
  • remove GFP promotor sequence*
  • remove DivivA stop codon*
  • Remove ComGA stabiliser sequence*
  • ‘Linkers’ were described by Edwards and Errington - we suspect that this is related to the restriction enzyme assembly process they used and hence didn’t include it in our design*

A sketch of design concept

Primer Design

used the Gibthon Construct Designer developed by the 2010 iGEM Cambridge team Left side of Div IVA sequence