Maintenance of the recombinant plasmid pIJ2 in chemostat cultures of Streptomyces lividans 66 (pIJ2)

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Abstract
The maintenance of the recombinant plasmid pIJ2 in chemostat cultures of Streptomyces lividans 66 (pIJ2) was investigated. The presence of the plasmid coding for a neomycin phosphotransferase was detected by plating samples from the continuous cultures on nonselective and selective agar medium containing neomycin.The plasmid was lost from the host strain under all conditions tested. However, the kinetics of the plasmid segregation from the chemostat populations were dependent on the growth‐limiting substrate of the medium, the dilution rate of the continuous culture and the cultivation temperature.Size differences between the original plasmid and plasmid DNA isolated from neomycin‐resistant clones after long‐term chemostat cultivation were not observed. In neomycin‐sensitive clones no extrachromosomal DNA was found.