In the life forms, the biomolecules such as DNA and protein are interacting with each other to maintain their life activity. Simultaneously, these biomolecules form DNA–protein complexes; as a result, the life forms can adjust to the external environment and continue its life activities. Therefore, using these characteristics of the DNA recognition ability of proteins, the novel molecular devices can be established. Here, we show the application of DNA binding and bending protein for design of the DNA actuator and its practical realization. The single polypeptide chain integrated host factor 2 (scIHF2), which is a DNA binding and bending protein, was used to bind to and/or bend the specific domains of DNA. Using this protein and designing the sequences of DNA, mechanical-functionalized DNA-based biomolecular device could be fabricated. Using these mechanisms, DNA binding and bending proteins have great potentials for establishing the DNA actuator for nanobiotechnology and nanotechnology applications.