As I want to study the interaction of two proteins in E. coli, I'm
looking for some advice. I already expressed one protein in E. coli and
now I'm up to the second protein.
My question is: Wchich is the most feasible method to make this proteins
interact in E. coli inner membranes? Is this coexpression by means of
compatible plasmids or polycistronic plasmids or even genomic integration
of one of the proteins, or might it be more intruiging to try to
reconstitute that interaction by fusion of phospholipid vesicles
(containing the purified second protein from a mammalian source) and E.
coli membranes (or even lysozyme treated bacteria). In the last case I
have the advantage that the second protein is posttranslational modified,
but it hasn't really been shown that this modification is necessary for
both proteins to interact.
Since this is a very critical decision to make, I'd really appreciate any
comment or suggestions.
Jurgen Vanhauwe, Ph. D. student