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From 2010.igem.org

Thursday 8th July - Meeting with Qaiser in the morning about the meeting in the afternoon. Unfortunately we couldn't do any more labs, as we're missing the DH5-alpha strain, but we had a funny email from Kate to lighten our moods. It read:

Hi all,

Hope you had a good gay yesterday - I'm still ploughing away with this work which I have to hand in tomorrow, but will be there tomorrow fingers crossed!

Hope it's all going good...

Kate

Note the spelling mistake. Yes, we are immature for laughing.

We worked through lunch ordering parts and plasmids. As we have not yet found the CqsA plasmid that 2008 used, we ordered another sample from the Bonnie Bassler institute, in the hope they'll once again be kind enough to supply us again. At 2pm we met with one of our microbial systems experts, Professor Jeff Green to talk about our idea and whether everything makes sense. He confirmed everything seems in order, but also liked our idea of trying the system two ways.

We should probably record how far the planning has gone - so here goes: Plan BarA is a sensory histidine kinase that sits in the E.coli cell membrane. CqsS is a cholera sensory histidine kinase, which detecs cholera autoinducers (quorum sensing molecules secreted extracellularly). We want to swap the outside sensory domain of the BarA protein with the outside domain with CqsS, then put the recombined BarA gene (with CqsS extracellular domain) into a high copy plasmid.

Meanwhile, we want to put a visible reporter, one of Cambridge's vibrantly coloured E. chromi protiens into the operon affected by the BarA internal kinase mechanism. This will be made in a BarA knockout strain (which we've ordered and hope to receive soon).

The alternative, which we will also give a go, is transforming the whole cholera CqsS system into E.coli, again with a visible reporter downstream.

When we arrived back from the meeting, productivity took a gradual decline and instead we finished the day discussing music and listening to Rhianna on youtube.