The invention discloses a structure and construction method of aperture steel plate shear keys. The structure comprises a steel beam composed of a steel beam flange plate (1) and a steel beam web (2), a T-shaped shear key is welded to the steel beam flange plate (1), the T-shaped shear key comprises a shear key flange plate (4) and a shear key web (5) with U-shaped grooves, each U-shaped groove and the steel beam flange plate (1) form a closed 'door hole' in which a main bar (6) is disposed, and the main bars (6) are provided with closed hoops (7). A concrete panel and the steel beam are integrated by using the T-shaped shear key, the structure not only has the advantages of simple structure, construction convenience and good integrity but also is good in pull resistance, high in longitudinal shear resistance and particularly applicable to hogging moment areas.