Cloning of the kanamycin resistance gene from a kanamycin-producing Streptomyces species

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Abstract
A kanamycin-producing strain, S. kanamyceticus ISP5500, is resistant to kanamycin. A kanamycin resistance determinant was cloned from S. kanamyceticus into S. lividans 1326, using the plasmid vector pIJ702. The resulting plasmid, pMCP5, could also transform S. lavendulae S985 and S. parvullus 2283 to kanamycin resistance. Transformants carrying pMCP5 were markedly more resistant than S. kanamyceticus to the aminoglycoside antibiotics sisomicin, tobramycin, amikacin and gentamicin. In vitro polyphenylalanine synthesis studies showed that strains carrying pMCP5 contained kanamycin-resistant ribosomes. Growing S. kanamyceticus contained kanamycin-sensitive ribosomes. Ribosomes from S. kanamyceticus grown under kanamycin-producing conditions were kanamycin resistant.