Team:KULeuven/24 July 2008

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Contents

Lab Work

Wet Lab

Stefanie and Hanne made a glycerolstock of all the parts we received from the iGEM headquarters. Everything is now stored in two -80°C freezers. You can find the cells with the parts in box S&PF7. You can look up in the cmpg database where you should search for a specific part. In the afternoon we started the protocol to make the right E.coli strain. We made a liquid culture of the donor and acceptor cells. Phage P1 was incubated with the acceptor. Tomorrow we'll continue the proces and bring phage P1 and the acceptor cells (MC0106) together, this should result in the right cells.

Dry Lab

Primers for the construction of T7 polymerase with UmuD tag were written out.

Modeling

Pulse generator has been reviewed, the parameters have been checked but there are issues... the pulse generator doesn't work (pure physically): when a step signal of T7 is applied cI is produced, which will then repress its transcription. Of course, this will develop to an steady state with a cI level equal to a non zero value.

A zero cI value would result in zero repression, thereby restarting the transcription of the cI gene and start producing cI again anyway.

Because the cI value remains non zero, so will the lactonase production, and so there will always be a constant amount of lactonase present. Question is how much this concentration will affect the system considering the case of no light present. No light present means HSL production which should bind to LuxR, but instead will react with lactonase to an hydroxy-acid. If the concentration of lactonase is considerable, and remains somewhat steady, cell death would never occur.

Sim pulse generator Lac.jpg Sim pulse generator cI.jpg

Interesting link to a pulse generator system that seems to work: Pulse Generator Keasling Lab

The memory system has been reviewed as well. The old system seemed unsuccesful, a new system has been found proven to work. There are however not enough repressors available to implement this system without having interference with other parts. ...

Wiki has been updated a bit more. SBMLSqueezer was used to produce pdf files with the celldesigner reactions, species and parameters. Biobricks pictures were added and some other stuff.

Remarks

iGEM gave us plasmids in cells. Which cells?

Quote of the day

GAS!