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[Qundo12]

I'm working with the pUB110 (from S. aureus) derived vector but I've got failed with the electroporation of the plasmid into an isolated Staphyloccocus. Many things can be the culprit for this but the first thing in my mind is maybe the large size of the plasmid I constructed inhibits the plasmid replication in Staph. This is a shuttle vector having both pUB110 ori for Staph and pBR322 ori for E. coli. I constructed several genes on this vector, so now I have 2 plasmids with the size of 15kb and 18kb.
Is there anyone here had experience dialing with this vector in Staph? Do you know what is the maximum length of DNA it can control?
I really appreciate any reply. Thank you all!

Edited by Quasimondo, 24 August 2009 - 06:35 AM.