Molecular cloning and characterization of protective outer membrane protein P.69 from Bordetella pertussis.

• DOI: 10.1073/pnas.86.10.3554
• Publication Date: 2006-05-31T10:55:09Z
• Authors: I G Charles; G Dougan; D Pickard; S Chatfield; M Smith; P Novotny; P Morrissey; N F Fairweather
• Keywords: Bordetella; Cosmid; Stop codon; Adenylate Cyclase Toxin

Protein P.69 is localized on the outer membrane of Bordetella pertussis and one virulence factors believed to contribute disease state whooping cough. We demonstrate that protein synthesis under genetic control vir locus. Using oligonucleotide probes derived from sequence a cyanogen bromide fragment, we have cloned gene for B. CN2992. Analysis DNA reveals G + C-rich capable encoding 910 amino acids with Mr 93,478, suggesting processed form larger precursor. In common some genes in toxin operon, CCTGG was found 5' ATG initiation codon. At 3' end, 29 bases after TAA stop codon, GTTTTTCCT may function transcription termination. A full-length clone carried by cosmid pBPI69 unable direct expression an Escherichia coli host. The generation P.69-fusion products allowed detection P.69-specific synthesized E. coli.