In this study, antimicrobial resistance of E. coli isolated from domestic beef on sale in Busan and Gyeongnam province was investigated from March to October 2008. A total of 400 beef samples were collected for the monitoring of antimicrobial resistance, and 39 (9.8%) strains of E. coli were isolated. Antimicrobial resistance test was carried out by agar disc diffusion method with 17 antimicrobials. In general, E. coli isolates showed the highest antimicrobial resistance to tetracycline (85.3%), followed by doxycycline (76.5%), streptomycin (61.8%) and sulfamethoxazole/trimethoprim (61.8%). Then they showed higher resistance to several antimicrobials like kanamycin and neomycin (55.9%). However, They had low antimicrobial resistance to amikacin (8.8%), amoxicillin/clavulanic acid (2.9%). Of 39 isolates, 31 (79.5%) were resistant to more than 2 antimicrobials. Among 17 antimicrobials examined, tetracyclines were the most resistant, followed by aminoglycosides, sulfonamides. The resistance was seemed to be correlated to amounts of antimicrobial use. In the result of this study, we suggest that there be need to regulate the abuse of antimicrobial on food-producing animals in Korea because the concern on antimicrobial resistant is gradually increased worldwide.