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Yes, I understand that long-term stability might be an issue. I also
realize that copy number control of the individual plasmids is something
to consider.
Thanks for the caution. I am also in the process of making of the dual
expression construct, which for this particular pair of proteins is
taking longer than I thought.
Raji
Artem Evdokimov wrote:
Two plasmids bearing same origin of replication will not co-exist in
the cell for very long
* although the above is correct, using plasmids that are in principle not
* compatible
* but have different resistance to antibiotics, works well in a number of
* cases and is easy to do.
Yes, I agree - this often does work for a while - which is why I wrote that
the two plasmids won't coexist for very long. So if you're only interested
in quick-and-dirty screening, you might want to try this. I would not
recommend this method for any work that requires long-term stability and
reproducibility of expression - the two plasmids of the same origin, that
bear different dominant selection markers, will eventually recombine into
one or more new plasmids (concatemers or recombinants bearing both markers)
and it's hard to say what will happen with respect to expression.
Artem
