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Team:Cambridge/Experiments/Initial Exercise Group One
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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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Revision as of 23:05, 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
Left side of the Div IVA coding sequence(site 1)
used the Gibthon Construct Designer developed by the 2010 iGEM Cambridge team
