plasmid rescue (plasmids with incompatible ORI) - (Jul/07/2009 )
I'm trying to ligate two plasmids (one with kanamycin and other with ampicillin selection markers) containing similar (pMB1 and ColE1) ORI's. I think as they fall in the same plasmid incompatibility group, I don't get any transformants on LB Kan/Amp selection plate. I'm using DH5 alpha chemically competent cells for transformation. Can someone please suggest if there is any bacterial strain where in a plasmid with two similar ORI's can exist? Or is there any other way to get this work done.
Thanks a lot
I don't think there is a way to get two plasmids of the same ORI to exist together..
Instead of reinventing the weal, ask around if anyone has a pCR vecter from Invitrogen. It has both kanamycin and ampicillin resistances.