Team:Cambridge/Experiments/Initial Exercise Group One

From 2011.igem.org

(Difference between revisions)
(Primer Design)
(Primer Design)
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*successful divIVA-GFP fusion documented by Edwards and ERrington, 1997*
*successful divIVA-GFP fusion documented by Edwards and ERrington, 1997*
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===Primer Design===
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===Construct Design===
We wish to:
We wish to:
*keep the original promotor for the Div IVA*
*keep the original promotor for the Div IVA*

Revision as of 23:00, 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