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Pseudomonas => E. coli expression...

Michael Coyne mjcoyne at warren.med.harvard.edu
Wed Jul 20 15:34:32 EST 1994


In article <1994Jul18.133039.1 at molbiol.ox.ac.uk>, rhubner at molbiol.ox.ac.uk
wrote:

> hi fellow bionetters,
>  does somebody know about expressing Pseudomonas genes under their own 
> promotors in E. coli? we have no antibodies for our gene at hand and it seems 
> -from a quick literature check- that Pseudomonas promoters are badly
> recognized IN GENERAL...
>  Thanks for any hint re:efficient systems... or approaches to avoid!
>   Roland


Roland...

Wellll...  What usually happens is that if it works, it's not a problem,
and if your gene is not expressed in E. coli, you can always blame it on a
missing upstream sequence :).

This topic actually comes up alot in here.  We know *some* genes are
efficiently expressed in E. coli (see, for example, Goldberg, et. al. 1992.
Cloning and surface expression of Pseudomonas aeruginosa O antigen in
Escherichia coli.  PNAS 89:10716-10720), and other P. aeruginosa genes
appear not to be (see Coyne, et. al. 1994. The Pseudomonas aeruginosa algC
gene encodes phosphoglucomutase, required for the synthesis of a complete
lipopolysaccharide core. J. Bacteriol. 176:12).  In the last paper (a
shameless plug), see the section on the complementation of the E. coli pgm
mutant with the P. aeruginosa gene.

For a more general review, see V. Deretic, et al. 1989. Common denominators
of promoter control in Pseudomonas and other bacteria. Bio/Technology.
7:1249-1254.  A bit old, perhaps, but still useful.

There are, of course, other methods of detecting protein production than
the use of antibodies.  Minicell and or maxicell analysis spring to mind.

Good luck...


Mike



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