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| Recombination Porcine Pm MetQ Induced Immune Protective Efficacy Against Challenged Infection of Pasteurella multocidain in Mice (Mus musculus) |
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Abstract Porcine (Sus scrofa) Pasteurella multocidain (Pm) infection caused serious diseases including swine plague, hemorrhagic septicaemia and infectious atrophic rhinitis. The present inactivated vaccine had poor cross protection effect and short immune protection period, and the recombinant vaccine still did not provide high protection rate. Based on Pm methionine transporter Q (metQ) sequences in GenBank, the primers were designed after removing the corresponding nucleotide sequence of expressing hydrophobic amino acid regions to express metQ in Escherichia coli. The partial sequence of Pm metQ was amplified by PCR from Pm genome. Bioinformatics analysis showed that the Pm metQ gene was composed of 768 bp, encoding a protein of 255 amino acids. This protein was an extracellular toxic protein and high homology among different Pm serotypes. Pm metQ gene was subcloned into pET32a (+) to construct pET-metQ. After induced by isopropylβ-D-1- thiogalactopyranoside (IPTG) rMetQ expressed in E.coli BL21, and determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that rMetQ with 28 kD was detected. Animal experiment was conducted by Kunming mice (Mus musculus). The serum immunoglobulin G (IgG) level was determined by enzyme-linked immuno sorbent assay (ELISA). The results showed that serum IgG level significantly increased in immuned group (aluminium adjuvant+rMetQ) at 14 d after third immunization. The infected mice in control groups were 20% survival rate at 3 d and all died at 7 d. The mice in the immunized group (aluminium adjuvant+30 μg rMetQ) showed 40% survival rate at 3 d and 10% survival rate at 7 d. But the mice in the immunized group (aluminium adjuvant+50 μg rMetQ) showed 80% survival rate at 3 days and 40% survival rate at 7 d. Autopsy of dead mice showed that the liver was the main diseased organ with haemorrhagic spot and even necrosis.Mice subcutaneously vaccinated with rMetQ revealed a partial protection against the challenge infection of Pm. This study provides data for screening new molecular vaccines against Pm infection.
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Received: 15 March 2017
Published: 30 October 2017
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