Expression and Purification of Outer Membrane Lipoprotein OprF in Recombinant Pseudomonas aeruginosa,and Preparation and Identification of Polyclonal Antibody

doi:10.13560/j.cnki.biotech.bull.1985.2015.08.031

Abstract

Abstract: The outer membrane lipoprotein of Pseudomonas aeruginosa（OprF）could antagonize bacterial infection by efficiently activating body immunologic mechanism, and therefore has an important perspective in vaccine development. The expression strain for OprF protein of P. aeruginosa was obtained by molecular clone;OprF protein was purified by the way of SDS-PAGE gel extraction and urea gradient renaturation, and the purified protein was used to immunize mice to prepare the polyclonal antibody. The titer of OprF antibody was 1∶12 800 according to ELISA, and Western blotting proved that the antiserum had good specificity. Homology analysis of OprF by DNAMAN showed that the C-terminal regions shared high homology in different bacteria. Phylogenetic analysis by MEGA revealed that genetic relationship with Pseudomonas genus bacteria was higher than that with others.

Key words: OprF protein, polyclonal antibody, ELASA, Western blotting, phylogenetic analysis

Chen Chunlin, Liu Xiang, Ju Xiong. Expression and Purification of Outer Membrane Lipoprotein OprF in Recombinant Pseudomonas aeruginosa,and Preparation and Identification of Polyclonal Antibody[J]. Biotechnology Bulletin, 2015, 31(8): 213-218.

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