Roof joint is a critical connection in space truss structures, which connects the upper steel space truss roofs and reinforced concrete structures. Because of its simple structural detailing and easy construction, a typical kind of roof joint is comprised of four embedded anchor rods, a mortar layer, a base plate, and stiffeners are popular in small- to medium-span space truss structures. Among them, base plates with slotted bolt holes are commonly used to release the deformation caused by temperature change or earthquake input. The roof joints were considered either as pinned or rigid connections in the design. In this paper, quasi-static tests investigating the influence of the applied axial force and the shape of bolt holes on the shear resistance and seismic behavior of roof joints were performed. The roof joints exhibited semi-rigid connection behavior and two types of initial resistance mechanisms, sliding and overturning, were observed. With the increase of applied axial force, the behavior of roof joint changes from sliding to overturning. Finally, effective methods to determine the initial resistance mechanism and calculate the ultimate shear strength of roof joints are proposed. These findings and analyses may provide guidelines for roof joint design practice for space truss structures.