Co-expression of different proteins in Escherichia coli using plasmids with identical origins of replication

It is generally accepted that the use of two different plasmids with the identical origins of replication in bacteria is not desirable due to their “incompatibility”. The utilization of the same bacterial enzymatic apparatus for replication of different plasmids is thought to cause a significant redistribution in favor of one of them. In the present work, examining co-expression of two different fluorescent proteins in Escherichia coli, we have shown that the use of highly homologous plasmids with identical origins of replication and providing resistance to different antibiotics results in high representation of both plasmids in bacteria. Meanwhile, the level of gene expression and the amount of proteins produced may differ and is determined mostly by their sequence rather than by the “incompatibility” of the plasmids.