In this study, we investigated relationship of muscular activity in thigh muscle and knee joint angle from bicycle exercise. The EMG signals of 16 persons were measured from rectus femoris and vastus lateralis muscles. The experiment was performed in 5 steps according to saddle distance and 60RPM/200W loads were applied for 1 minute at each step. EMG activation of rectus femoris and vastus lateralis muscles and knee joint angle were recorded using surface EMG and motion analysis system, respectively. Experimental results of inter relationship between EMG activation of thigh muscles and knee joint angle showed high correlation from Step 1. The unified EMG activation of two muscles and knee joint angle showed negative correlation(-0.97).