Abstract: As an important virulence factor of Brucella, lipopolysaccharide（LPS） plays a significant role in the process of Brucella infecting the host. Based on the bioinformatics analysis of the important proteins related to the synthesis of Brucella LPS, this study selected the key gene per of the Brucella LPS O side chain for cloning and expression, aiming to explore the function and application potential of Brucella LPS. According to the results, the Brucella LPS synthesis-related 30 important proteins are mainly hydrophilic proteins without signal peptides and transmembrane helix structures, all of which have strong reactogenicity and methylation and phosphorylation modification sites. Among them, 18 proteins（18/30） can be modified by glycosylation, and 4 proteins（4/30） can be modified by acetylation. Subcellular location showed that most of the proteins function in the cytoplasm, and a few of them function in the periplasm and the inner membrane of the cell. In addition to the Wbpw protein, there can be interactions between important proteins related to the synthesis of Brucella LPS. The pET28 a-per recombinant expression plasmid was constructed, and 45.7 kDa protein was expressed in the supernatant after induction. Through the Western Blot, the protein was verified to be highly reactive. For the follow-up research on the function of Brucella LPS and the research and development of new vaccines as well as diagnostic technologies, this research is considered a scientific foundation.