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Aminoglycoside antibiotics are among the oldest,and the most clinically important aminosugars primarily isolated from the various Streptomyces and Micromonospora.This study is mainly focused on the establishment of heterlogous expression of the biosynthetic gene cluster for the identification of aminoglycoside genes and expression of the recombinant minimal sets of candidates genes to prove the biosynthetic pathway of kanamycin [1-2].2-Deoxystreptamine and paromamine are two key intermediates in kanamycin biosynthesis.Two gene cassettes were constructed and expressed in Streptomyces fividans TK24.The expression of first set of cassette produced kanamycin B along with kanamycin C,where as only kanamycin C was produced from the second set of cassette.