activity of mammalian promoters in E.coli?

Chris Baumann baumannc at pop.nci.nih.gov
Tue Nov 11 07:46:46 EST 1997


Are there any E. coli promoters in the vector (i.e. from the AmpR or B-gal) which read
in the same direction as your clone.  You will often get low levels of read through txn
from these promoters and if your gene is indeed  toxic, this may be enough to kill your
cells.

Stephan Kuppig wrote:

> Dear Netters,
>
> the problem I faced recently was like this: I attempted to clone the cds of my
> protein of interest into a mammalian expression vector and failed to get
> positive E.coli clones. This protein (if expressed) is potentially toxic for
> bacteria - even in small amounts. Does anybody  know wheter the human beta
> actin promoter (that´s the one my expression vector has) can drive expression
> in E.coli? And what about other promoters  (e.g. CMV)? Any suggestions are welcome!
>
> Thanks in advance
>
> Stephan
>
> Stephan Kuppig
> Max-Planck-Institute for Immunobiology
> Stuebeweg 51
> D-79108 Freiburg, Germany
>
> Email: kuppig at immunbio.mpg.de






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