A new method of plasmid DNA transfer from the donor strainEscherichia coliS17-1 to the erythromycin-producing strainSaccharopolyspora erythraeaand avermectin-producing strain Streptomyces avermitilis via intergeneric conjugation was proposed. The optimal parameters of the method were chosen for increasing the efficiency of crosses and ensuring easily reproducible results. The behavior of the multicopy plasmid pPM803 and the integration vector pTO1 along with a number of new plasmids specially created by us, was examined in these strains. A new plasmid vector (pSI60) capable of integrating into the chromosome of actinomycetes at the integration site of the temperate actinophage φC31 was constructed. This vector possesses unique sites convenient for cloning and may be stably maintained in exconjugants of S. avermitilis and in the model strain Streptomyces lividans.The gene encoding resistance to spectinomycin and streptomycin was cloned into the vector pSI60 in this strain. For cloning in strain Sac. erythraea, vectors pSI261–280, which integrate into the chromosome via homology with the cloned DNA and can be stably maintained in exconjugants, were constructed.
Russian Journal of Genetics – Springer Journals
Published: Oct 7, 2004
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