Pasteurella Multocida Outer Membrane Proteins And Secreted Proteins Protective Antigen Screening

Swine Pasteurella multocida disease is a kind of common disease and still have not been solved now in farms, causing economic losses for pig farms. There is an urgent need for a stable and reliable vaccine to prevent the disease.The experiment subcellular localizated more than 2000 open reading frame gene of Pasteurella multocida serum type D. We screened Forty-six genes, including Thirty-nine protein outer membrane protein, seven protein is secreted protein.After PCR amplificated the gene and transfered it into plasmid and E. coli expression, Thirty-six protein were expressed, include five soluble protein, and the inclusion of 31 spheroidin protein.These 36 proteins were formulated into a mouse vaccine, and 7 proteins were found to be protective. Omp87 protein, as a kind of outer membrane protein, was confirmed by foreign scholars for a certain protective, this experiment carried out the construction of the protein gene and the successful expression. It was found that the Omp87 protein has a protective rate of 40%.hypothetical protein(NO.1), Omp87-1(NO.42), HasR protein(NO.46) have a 40% protection rate, and hypothetical protein (NO.40), Omp87-2 (NO.43),TonB-dependent receptor(NO.47) protein has a 20% protection rate.By after the first immunizated 13d and the second immunizated after 13d with whole serum collected bacteria coated 96-well ELISA plates to detect antibodies. Found that value in addition to a free 13d vaccine group other groups relatively low. After two free detection antibody rised after 13 d, in which positive antibody vaccine group the highest value, commodity groups, and Omp-87-1(No.42) groups and TonB-dependent receptor (No.47) on the revaluation is relatively high, and the hypothetical protein (No.40), Omp87-2 (No.43), HasR protein (No.46) antibody is relatively low, compared to the negative control than the obvious difference.By western blot experiment we found that these protein can reacted with swine Pasteurella multocida positive serum.All there are seven protein immune mice can against Pasteurella multocida affection. These protein can be used as a candidate antigen of Pasteurella multocida.