The purpose of this study was to identify the effect of the hip internal rotation on gluteal and erector spinae muscle electromyographic (EMG) activity during treadmill walking. Eleven healthy subjects were recruited. All subjects performed treadmill walking while maintaining the hip in neutral position (condition 1) and in internal rotation (condition 2). Surface EMG activity was recorded from four muscles (gluteus maximus (GM), gluteus medius (GMED), tensor fascia latae (TFL), and erector spinae (ES)) and the hip internal rotation angle was measured using a three dimensional motion analysis system. The gait cycle was determined with two foot switches, and stance phase was normalized as 100% stance phase (SP) for each condition using the MatLab 7.0 program. The normalized EMG activities according to the hip rotation (neutral or internal rotation) were compared using a paired t-test. During the entire SP of treadmill walking, the EMG activities of GM in condition 1 were significantly greater than in condition 2 (p<.05). The EMG activities of TFL and ES in condition 2 were significantly greater than in condition 1 (p<.05). The EMG activities of the GMED in condition 1 were significantly greater than in condition 1 (p>.05) except for 80~100% SP. Further studies need randomized control trials regarding the effect of hip internal rotation on the hip and lumbar spine muscle activity. Kinetic variables during gait or going up and down stairs are also needed.