The paper attempts to research the seismic behavior and mechanical mechanisms under repeated low-cycle loading of vertical connections in prefabricated reinforced concrete structures. Eighteen specimens were designed and tested, the test process and the mechanical mechanisms are studied, the seismic behavior of connections is analyzed, and the shear resistance formulas are proposed. The theoretical values based on proposed formulas are good agreement with test results. Under repeated loading, the shear resistance decreases, the connection ductility increases with connecting bar, but decreases with connection width. However, all the connections fail crisply with poor ductility. At sudden-cracking, the shear resistance increases with connecting bar, but changes nonlinearly with connection width. The resistance is composed of attributions of mechanism of diagonal-compressive column (MDCC) and mechanism of compressive friction action (MCFA). After sudden-cracking, the shear resistance is provided by weakened MDCC and dowel action of connecting bars (DACB). It increases with connecting bar and connection width.