It'd more be a proof of concept, since you can have toxic protein stable under the rhamnose promoter.
-- If it works well enough, I'll switch it out for a cl/cro system or a flp system so you only have to induce once.
-Koeng
On Sunday, March 22, 2015 at 10:21:05 PM UTC-7, Dakota wrote:
On Sunday, March 22, 2015 at 10:21:05 PM UTC-7, Dakota wrote:
Rhamnose seems more expensive than common antibiotics for selection , or at the very least, compatible.
On Mar 22, 2015 8:55 PM, "Koeng" <koen...@gmail.com> wrote:The immunity mechanism actually (hypothetically) involves blocking binding to an inner membrane protein sdaC. TolC is used in conjunction with cvaA/B for export--I have an experiment planned which I think will work for DIY transformations, will update if it works. Essentially, I want to integrate the ColV operon without the immunity protein into the genome under the rhamnose promoter. Upon activation of rhamnose, all cells should die. Or alternatively they can have the cvi protein on a plasmid to survive. This entire experiment should only take 2 cloning reactions, which is nice. Quick to complete, relatively.-Koeng
On Sunday, March 22, 2015 at 2:56:15 AM UTC-7, Mega [Andreas Stuermer] wrote:According to wikipedia, one molecule was speculated to be able to kill a cell. However, pUC Ori copy number is 100 to 500 per cell and so you should have tons of colV to kill other cells.
I haven't looked into the immunity mechanism but it involves blocking the TolC receptor ehere colicinV usually binds too.
You could definitely try to induce colV-immunity with IPGT to see if that increases survival.
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