The provincial laboratories are scientific and technological innovation platforms built by the provinces for national strategic objectives and provincial strategic needs, focusing on major scientific frontiers, scientific and technological tasks, and critical core technology breakthroughs. First, this study analyzes the development process and construction situation of provincial laboratories in China. It concludes that the provinces adhere to the “two-wheel drive” construction model of scientific and technological innovation and institutional innovation. More than 100 provincial laboratories have been established and constructed in China, and a series of construction and operation experiences and institutional innovation achievements have been achieved. Secondly, based on innovation ecosystem theory, this study summarizes the essential elements involved in the construction system of provincial laboratories from four aspects: innovation goals, innovation subjects, innovation networks, and innovation environments. Combined with PDCA cycle management theory, this study analyzes vital problems to be solved in the construction system of provincial laboratories. Finally, the current dilemma of provincial laboratories construction is analyzed, and the provincial laboratory’s high-quality development and national science and technology innovation system optimization can be effectively promoted through means such as optimizing top-level design, strengthening subjects’ responsibility, improving management system, innovating operation mechanisms, improving evaluation system and other ways.