I would like to ask for the advice of those of you who had the experience
with expressing plant chloroplast envelope proteins in E. coli.
* Does a predicted transit peptide need to be removed from the E. coli
* Do codons need to be modified to reflect optimal E.coli codon usage?
* Would the plant protein be targeted into the E.coli cell membrane by
does a signal sequence need to be added?
* Is there any reliable way to predict the topology of the protein in the
i.e. which way (in or out of the cell) the N- and C-termini would face?
* If the chloroplast inner membrane protein works as a transporter and the
hypothesis is that in plants it pumps the substrate out of the plastid, is
way to predict the direction of substrate flow in E. coli (assuming the
is functional in bacteria)?
* If the plant chloroplast protein (minus the transit peptide) is
yeast, does all of the above hold true?
Thank you for your help,