A simple method for construction of pir+ Enterobacterial hosts for maintenance of R6K replicon plasmids.

ABSTRACT:

Background

The R6K replicon is one of the best studied bacterial plasmid replicons. Replication of the R6K plasmid and derivatives harboring its γ origin of replication (ori(R6Kγ)) is dependent on the pir gene-encoded π protein. Originally encoded by R6K, this protein is usually provided in trans in hosts engineered to support replication of plasmids harboring ori(R6Kγ). In Escherichia coli this is commonly achieved by chromosomal integration of pir either via lysogenization with a λpir phage or homologous recombination at a pre-determined locus.

Findings

Current methods for construction of host strains for ori(R6Kγ)-containing plasmids involve procedures that do not allow selection for presence of the pir gene and require cumbersome and time-consuming screening steps. In th

SUBMITTER: Kvitko BH 

PROVIDER: S-EPMC3338088 | biostudies-literature | 2012 Mar

REPOSITORIES: biostudies-literature