Team:Cambridge/Experiments/Initial Exercise Group One

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(Primer Design)
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===Primer Design===
===Primer Design===
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====Left side of the Div IVA coding sequence(site 1)====
used the Gibthon Construct Designer developed by the 2010 iGEM Cambridge team
used the Gibthon Construct Designer developed by the 2010 iGEM Cambridge team
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[[File:Cam_Oligo design_site1.png|frameless|700px|Left side of Div IVA sequence]]
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[[File:Cam_Oligo design_site1.png|frameless|900px|Left side of Div IVA sequence]]
{{Template:Team:Cambridge/CAM_2011_TEMPLATE_FOOT}}
{{Template:Team:Cambridge/CAM_2011_TEMPLATE_FOOT}}

Revision as of 23:05, 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

Left side of the Div IVA coding sequence(site 1)

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