Mycoplasma pneumoniae (M.pneumoniae), one of the smallest and simplest cell wall-deficient bacteria that can cause community-acquired pneumonia (CAP).Vaccination is a possible way to reduce the infection effectively.The transmembrane protein P116N(N-terminal of protein), P1C(C-terminal of protein), P30, and the chimeric protein P116N-P1C-P30 (MP559) were expressed and purified by genetic engineering respectively.The four proteins were respectively synergized with freund adjuvant for immunizing New Zealand white rabbits three times, including boosted immunization twice and shock immunization once.The IgG antibodies in the sera of immunized rabbits were detected by western blot and ELISA to analyze the antigenicity and immunogenicity.