Team:Cambridge/Experiments/Initial Exercise Group One

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