Team:Cambridge/Experiments/Initial Exercise Group One

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OVERVIEW
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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*

Primer 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