Private higher education in China has gradually adapted to marketing economy and entered into a benign circle of self-support and self-development. As an important part in the management of private universities, human resource construction has become a great concern. Motivation is of great importance in the construction. This paper starts from the reality of private universities, and studies the application of motivation mechanism through literature survey and empirical research. Firstly, the concept of motivation and the operating mode of motivation mechanism are made clear in the paper. Then, the current situation of motivation in private universities is analyzed to explore the potential problems. Finally, suggested measures like construction of fair system, integrated application of different motivation mechanisms, establishment of communication channels, and combination of individual motivation with group motivation, are put forward to provide reference for the human resource construction in private universities.