To find out the main excitation sources of a bus floor's vibration, modal analysis and spectral analysis were respectively performed in the paper. First we tested the vibration modal of the bus's floor under the full-load condition, and the first ten natural frequencies and vibration modes were obtained for the source identification of the bus floor's vibration. Second the vibration characteristic of the bus floor was measured in an on-road experiment. The acceleration sensors were arranged on the bus's floor and the possible excitation sources of the bus, which includes engine mounting system, driveline system, exhaust system, and wheels. Then the on-road experiment was carefully conducted on a highway under the four kinds of test condition: in-situ acceleration, uniform velocity (90km/h, 100km/h, 110km/h, 120km/h), uniform acceleration with top gear, and stall sliding condition with neutral gear. After that, by performing order tracking analysis and spectral analysis, the 1st order rotation frequency of the driveline and the 2nd order frequency of engine were identified to be the main cause of bus floor's vibration. At last, the reasonable measures were proposed to reduce the vibration of the bus's floor.