To detect the immune effectiveness of Pseudomonas aeruginosa oprI gene DNA vaccine and its recombinant subunit vaccine.The target gene fragment was cloned into the eukaryotic expression vector pCAGGS-HA to construct recombinant plasmid pCAGGS-oprI.After enzyme digestion,the DNA vaccine was obtained.At the same time,the oprI gene fragment was cloned into the prokaryotic expression vector pET32a to construct recombinant plasmid pET32a-oprI.Then the recombinant plasmid was identified by digestion and transferred into Escherichia coli induced with IPTG.After the recombinant protein was expressed and purified by affinity chromoatography,the recombinant subunit vaccine was prepared and tested with SDS-PAGE.BALB/c mice were vaccinated with DNA vaccine,recombinant subunit vaccine,and outer membrane vaccine and inactivated vaccines as the controls.Then sera were collected every 7 days,and the levels of antibody,interferon-γ(IFN-γ),interleukin-2(IL-2)and interleukin-4(IL-4)concentrations were detected by indirect ELISA.The mice were challenged with virulent Pseudomonas aeruginosa following primary immunication for 42 days,and protective efficacy of vaccines were evaluated.The results showed that the serum antibody and cytokine levels in the four vaccine groups were significantly higher than those in the negative control group(P＜0.05).The serum antibodies level and IL-4 concentration in the recombinant subunit vaccine group were higher than those in the DNA vaccine group(P＜0.05).No significant differences were detected for the concentrations of IFN-y and IL-2 between the DNA vaccine group and recombinant subunit vaccine group(P＞0.05).The protective efficacy rates of DNA vaccine group,recombinant protein vaccine group,outer membrane vaccine group and inactive vaccine group were 45％,55％,70％and 95％,respectively.The results indicated for the first time that both the DNA vaccine and recombinant subunit vaccine based on the oprI gene of Pseudomonas aeruginosa could induce immune response and provide immune protection from Pseudomonas aeruginosa infection in mice.However,The immune effectiveness of these two vaccines need to be enchanced further.Our study was to lay the foundation for the development of vaccine against the Pseudomonas aeruginosa.
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