Intergeneric conjugation between Escherichia coli and Streptomyces species

We have constructed Escherichia coli-Streptomyces shuttle plasmids which are capable of conjugal transfer from E. coli to Streptomyces spp. These plasmids contained the pBR322 and pIJ101 origins of replication and the RK2 (IncP) origin of transfer. The transfer of plasmid was specifically dependent the presence of a 760-base-pair, cis-acting, oriT-containing fragment and on RP4 (IncP) functions supplied in trans. Conditions of mating and selection of exconjugants were analyzed with Streptomyces lividans as recipient. Plasmid transfer to other Streptomyces species was also demonstrated.