Abstract: Acidithiobacillus ferrooxidans lives in aerobic environments where concentration of iron is extremely high and the pH is low. It can synthesize intra-cellular electron-dense magnetite, called magnetic-particle，which can be extensively used as bio-nano-materials. In order to explore the influence of oxygen on magnetosome formation in Acidithiobacillus ferrooxidans, the different shaking frequency responses of feoB, mpsA, tonB and ABC gene were analyzed by using real-time PCR. Transmission electron microscopy was also used to analyze the magnetic-particle formed by this bacterium in different shaking frequencies. The result shows that the oxygen has some effect on the expression of the genes and the formation of the magnetic-particle, which lays the foundation for the deep research of the mechanism of the formation of magnetic-particle.