Dynamic tests of three bonded and two unbonded full-prestressed concrete beams were carried out. The purpose was to seek the relation between prestressing force and natural frequency.Test results indicate that the frequency of prestressed concrete (PSC) beam increases with the increase in prestressing force approximately. The results are different from the dynamic characteristics of isotropic material beam subjected to compressive axial force which were put forward by Clough et al. The reason is that the beams were considered as isotropic, homogeneous, linear elastic material in the traditional analysis method. However, more accurate results are required in the analysis of frequency of PSC beam. The constitutive mode of PSC member is analyzed based on microstructure of concrete in this paper. The orthotropic linear elastic mode is used to analyze the relation between dynamic frequency and prestressing force of concrete beam, at the same time the equivalent stiffness of prestressed tendon relating to the prestressing force is added to the bending deformation stiffness of the beam. The analytical value agrees well with the test result, indicating that the current analysis method in this paper is feasible to full-prestressed concrete beam.