Kanamycin acetyltransferase gene from kanamycin-producingStreptomyces kanamyceticus IFO 13414

A kanamycin producer,Streptomyces kanamyceticus IFO 13414 is highly resistant to kanamycin. Cloning of the kanamycin resistance genes inS. lividans 1326 with pIJ702 gave several kanamycin resistant transformants. Two transformants,S. lividans SNUS 90041 andS. lividans SNUS 91051 showed similar resistance patterns to various aminoglycoside antibiotics. Gene mapping experiments revealed that plasmids pSJ5030 and pSJ2131 isolated from the transformants have common resistant gene fragments. Subcloning of pSJ5030 gave a 1.8 Kb gene fragment which showed resistance to kanamycin. Cell free extracts ofS. lividans SNUS 90041,S. lividans SNUS 91051 and subclone aS. lividans SNUS 91064 showed kanamycin acetyltransferase activity. The detailed gene map is included.