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