A novel precast concrete-encased steel composite beam, which can be abbreviated as PCES beam, is introduced in this paper. In order to investigate the shear behavior of this PCES beam, a test of eight full-scale PCES beam specimens was carried out, in which the specimens were subjected to positive bending moment or negative bending moment, respectively. The factors which affected the shear behavior, such as the shear span-to-depth aspect ratio and the existence of concrete flange, were taken into account. During the test, the load-deflection curves of the test specimens were recorded, while the crack propagation patterns together with the failure patterns were observed as well. From the test results, it could be concluded that the tested PCES beams could all exhibit ductile shear behavior, and the innovative shear connectors between the precast concrete and cast-in-place concrete, namely the precast concrete transverse diaphragms, were verified to be effective. Then, based on the shear deformation compatibility, a theoretical model for predicting the shear capacity of the proposed PCES beams was put forward and verified to be valid with the good agreement of the shear capacities calculated using the proposed method and those from the experiments. Finally, in order to facilitate the preliminary design in practical applications, a simplified calculation method for predicting the shear capacity of the proposed PCES beams was also put forward and validated using available test results.