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

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