Abstract

Plasmids containing the same origin of replication (pBBR1MCS-2 KmR and pBBR1MCS-3 TcR) have been used to express simultaneous and independently different proteins in P. putida U and in E. coli. Thus, when P. putida was transformed with different genetic constructions made in the same plasmid (pBBR1MCS-3 TcR), or with plasmids containing the same replication origin but with different antibiotic resistant genes (KmR and TcR), they co-existed inside the same microbe. Furthermore, when E. coli DH10B was transformed with the plasmids recovered from the recombinant P. putida U, we noticed that all the bacteria isolated from single colonies are resistant to Km and Tc, suggesting that these plasmids were also present in E. coli. This observation facilitates the genetic manipulation of these strains (i.e. avoiding the use of different plasmids in double or multiple complementation experiments), and could be an interesting tool to approach many metabolic and biotechnological studies.

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