We have developed an efficient high yielding neomycin resistant Strain of Streptomyces fradiae for improved production of neomycin. Streptomyces fradie was submerged in the solution of10 ug/ml, 15 ug/ml, 20 ug/ml,30 ug/ml,40 ug/ml,50 ug/ml, 60 ug/ml,70 ug/ml, 80 ug/ml, for five day for screeing out high yielding Neomycin resistant Strain of Streptomyces fradiae . Streptomyces fradie in the solution of 70 ug/ml was the high yielding Neomycin resistant Strain of Streptomyces fradiae It was same observed as prevous authors that optimal levels of the elements Ca, Fe, and Zn per milliliter of a synthetic medium for neomycin production . K2HP04 was required at a concentration of 0.1% for maximal yield of neomycin but observed improved production of neomycin. It was observed 252 ug/ml, neomycin production.