low copy origin for expression plasmid

Tosso Leeb tleeb at gwdg.de
Thu Jan 22 05:28:54 EST 1998


Dear Netters,

I am trying to clone a large eukaryotic membrane protein into the =

mammalian expression vector pIRES-EGFP from Clontech. It looks like this =

construct is toxic for the E. coli because I can only isolate mutated =

DNA. My vector contains a ColE1 origin. I am thinking of replacing this =

high copy origin with a low copy origin to reduce the toxic effects on =

the bacteria. Does anyone have a suggestion which origin might be useful =

and where I can get DNA so that I can clone it? Any help is =

appreciated, thank you very much,
Tosso Leeb
-- =

Institute of Veterinary Medicine
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