title: Direct expression of Bordetella pertussis filamentous hemagglutinin in Escherichia coli and Salmonella typhimurium aroA.
abstract: Nonfused (i.e., nonhybrid) filamentous hemagglutinin (FHA) of Bordetella pertussis was efficiently expressed in Escherichia coli K-12 and Salmonella typhimurium aroA at levels higher than those found in wild-type B. pertussis when the upstream signals of the gene were replaced and the translation initiation region was engineered to optimize translational efficiency. Inclusion of part of the C-terminal FHA open reading frame, whose translation product does not appear to be part of the major secreted species of FHA, was shown to be important in achieving protein expression in both E. coli and S. typhimurium aroA; removal of the downstream gene sequence abolished recombinant FHA production. The levels of expression observed varied widely according to the construct and host bacterium used.
authors.affiliation: Department of Microbiology, GBF-National Research Center for Biotechnology, Braunschweig, Germany.
authors.forename: C A
authors.initials: CA
authors.lastname: Guzmán
authors.forename: M J
authors.initials: MJ
authors.lastname: Walker
authors.forename: M
authors.initials: M
authors.lastname: Rohde
authors.forename: K N
authors.initials: KN
authors.lastname: Timmis
mesh_terms: D018829:Adhesins, Bacterial; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D001886:Bordetella pertussis / Q000235:genetics*; D002874:Chromosome Mapping; D000939:Epitopes; D004926:Escherichia coli / Q000235:genetics*; D006388:Hemagglutinins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D014176:Protein Biosynthesis; D012486:Salmonella typhimurium / Q000235:genetics*; D010566:Virulence Factors, Bordetella
chemical_list: D018829:Adhesins, Bacterial; D000911:Antibodies, Monoclonal; D000939:Epitopes; D006388:Hemagglutinins; D010566:Virulence Factors, Bordetella; C065273:filamentous hemagglutinin adhesin, Bordetella pertussis