Hello Everyone,
I am developing a system with two plasmids. One of the plasmids have been assigned the incompatibility group of IncH, the other is plasmid pK19mobsacB in which I am looking into the group.
Assuming they have different incompatibility groups, would it be still proper or necessary for publication for me to check this? Keep in mind that I did insert a gene into pK19mobsacB, but it shouldn't effect its replication pattern.
If I can not assume that the two plasmids are compatible based off of the groups they have been assigned to, what would be the best way of testing their compatibility?
Thanks.
Plasmid incompatibility is typically only a problem when you need the two plasmids with the same group maintained stably over multiple growth cycles without antibiotic selection. It's typically not an instantaneous thing.
By the way, pK19, which I believe is the backbone plasmid, seems to have a pBR322 origin. That would make it a ColE1 plasmid. I can't remember which incompatibility group that is. Source: https://www.addgene.org/vector-database/3302/
Even incompatible plasmids will not be lost immediately if they are multi-copy. See here: https://academic.oup.com/peds/article/20/7/309/1546372/Plasmid-incompatibility-more-compatible-than
https://academic.oup.com/peds/article/20/7/309/1546372/Plasmid-incompatibility-more-compatible-than
Dear Adam,
It is very unlikely that you would observe an incompatibility reaction between your IncH-group plasmid and pK19mobsac (which probably contains the IncA-group origin of replication from pMB1 rather than the IncA-group replicon from ColE1 as suggested by Alexandra) in Escherichia coli.
Most significant incompatibility reactions result from problems associated from similar replicons (belonging to the same Inc group), but weaker incompatibility can result from similar partitioning and conjugation systems: none of these should effect your experiments as most engineered vectors such as pK19mobsac do not contain these additional plasmid maintenance functions. However, you should check to make sure that you are using different antibiotics to maintain selection for each plasmid during your experiments.
Regards, Andrew.
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