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Team:Cambridge/Experiments/Initial Exercise Group One
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Revision as of 23:08, 6 July 2011
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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*
Primer Design
used the Gibthon Construct Designer developed by the 2010 iGEM Cambridge team
Left side of the Div IVA coding sequence(site 1)
Right side of the Div IVA coding sequence(site 2)
