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| Immune Protection Research Against Challenge Infection in Mice (Mus musculus) Induced by APP rOmpP2 of Porcine (Sus scrofa) |
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Abstract Abstract Porcine contagious pleuropneumonia (PCP) is a respiratory disease caused by Actinobacillus pleuropneumoniae (APP) infection, which is one of the 5 major epidemics in the world's pig industry. Based on APP OmpP2 sequences in GenBank, the primers were designed. APP OmpP2 was amplified by PCR from APP genome, sequenced and submitted to NCBI (Access No. KM357903). Bioinformatics analysis showed that the APP OmpP2 gene was composed of 1 131 bp, encoding a 376 amino acid protein. This protein was a porin protein with high homology among different APP serotypes. APP OmpP2 gene was subcloned into pET32a (+) to construct recombinant plasmid pET-OmpP2. rOmpP2 expression in Escherichia coli BL21 was induced by isopropyl β-D-1- thiogalactopyranoside (IPTG), and determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS- PAGE) and Western blot. The results showed that rOmpP2 with 42 kD was detected. Animal experiment was conducted by BALB/c mice (Mus musculus). The serum immunoglobulin G (IgG) level was detected by enzyme-linked immuno sorbent assay (ELISA). The results showed that survival rate of infected mice with aluminium adjuvant+50 μg rOmpP2, aluminium adjuvant+30 μg rOmpP2 and aluminium adjuvant+10 μg rOmpP2 were 50%, 30% and 10%, respectively, and all mice died in adjuvant group and control group. The surviving mice slowly returned to normal health status. Mice that were subcutaneously immunized with rOmpP2 showed an partial protection against the challenge with APP. This study provides basic data for screening new molecular vaccines of porcine contagious pleuropneumonia.
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Received: 30 June 2016
Published: 13 January 2017
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