Recombinant Expression and Immunogenicity Analysis of the Porin Protein OmpF of Aeromonas hydrophila

doi:10.13560/j.cnki.biotech.bull.1985.2019-0628

Abstract

Abstract: Outer membrane protein（Omp）F in Aeromonas spp. is β-barrel like porin,and involves in the adaption of osmotic pressure and is highly conservative in this common pathogen for human and fish,thus poses a promising research value as immune-detection target. The A. hydrophila OmpF fragment published in GenBank was synthesized,and inserted into plasmid pET-28a（+）. The recombinant plasmid was transformed into Escherichia coli BL21（DE3）and induced by isopropy-β-D-thiogalactoside（IPTG）. The molecular size of the expressed protein was about 40 kD,which mainly expressed as inclusion bodies after optimization of inducing temperatures and IPTG concentrations. BALB/c mice were immunized with purified OmpF inclusion bodies. The results of ELISA showed that the immunized mice serum cross-reacted with 10 of 11 boiled and deactivated strains of Aeromonas spp.,the serum titers were around 1∶81 000. However,the mice serum did not react with Vibrio cholerae,Bacillus licheniformis,V. parahaemolyticus,V. vulnificus,and V. anguillarum. The above results indicate that the protein OmpF is immunogenic,conserved and was promising in developing Aeromonas cross-reactive antibodies.

Key words: Aeromonas hydrophila, outer membrane protein OmpF, recombinant expression, immunogenicity

GAO Yun-shan, LIU Dan-dan, XU Jun-lin, SANG Yu-nong, LIANG Xia-xia, LIU Jian-xin, WANG Wen-bin. Recombinant Expression and Immunogenicity Analysis of the Porin Protein OmpF of Aeromonas hydrophila[J]. Biotechnology Bulletin, 2019, 35(9): 234-243.

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