The purpose of this study was to isolate and characterize the Fusobacterium nucleatum (F. nucleatum) from subgingival plaque in Korean periodontitis patients. The subgingival plaque samples of periodontitis patient were collected with sterilized paper point. The paper point was put into reduced transfer medium and then immediately transferred to laboratory. The subgingival samples were diluted by 10,000 folds and plated on F. nucleatum-selective media agar plate. The plates were incubated at 37℃ in an anaerobic chamber for 3 days. The violet-colored colonies were selected and subjected to further verification whether those are F. nucleatum or not. For further confirmation, 16S rRNA genes (rDNA) were cloned from each of bacterial clones and determined sequence of 16S rDNA. In this study, we found 17 distinct clinical isolates of F. nucleatum from subgingival plaque. The clinical isolates will be a useful in various studies in periodontology.