Cloning and sequencing of the structural gene for the porin protein of Bordetella pertussis

Abstract

Bordetella pertussis produces a porin protein which is a prominent outer membrane component found in both virulent and avirulent strains. N-terminal amino acid analysis of purified B. pertussis porin was performed and this amino acid sequence was used to design an oligonucleotide that was then utilized to screen a lambda gt11 library containing randomly sheared fragments of DNA from B. pertussis strain 347. One clone, lambda BpPor, was identified and subcloned into pUC18. A portion of the DNA insert in this subclone, pBpPor1, was sequenced and shown to contain the N-terminal region of the structural porin gene. This truncated gene sequence was used to design an additional oligonucleotide that was used to identify a clone, pBpPor2, which overlapped with pBpPor1 and contained a termination codon. The structural gene deduced from this sequence would encode a 365-amino-acid polypeptide with a predicted mass of 39,103 daltons. The predicted product also contains a signal sequence of 20 residues that is similar to that found in other porin genes. The predicted B. pertussis porin protein sequence contains regions that are homologous to regions found in porins expressed by Neisseria species and Escherichia coli, including the presence of phenylalanine as the carboxy-terminal amino acid. DNA hybridization studies indicated that both virulent and avirulent strains of B. pertussis contain only one copy of this gene and that Bordetella bronchiseptica and Bordetella parapertussis contain a similar gene.

Membrane Localization of the S1 Subunit of Pertussis Toxin in Bordetella pertussis and Implications for Pertussis Toxin Secretion

“…The lack of solubility of the membrane-bound, full-length S1 subunit in Triton X-100 was similar to the lack of solubility of PtlF (Fig. 6B), a protein which is believed to localize to the outer membrane of B. pertussis (23), and to the lack of solubility of the major porin protein of B. pertussis (data not shown), which is known to localize to the outer membrane (31). In contrast, a significant fraction of PtlE, a protein believed to localize to the inner membrane (23), was soluble in Triton X-100 (Fig.…”

Proteome approaches combined with Fourier transform infrared spectroscopy revealed a distinctive biofilm physiology in Bordetella pertussis

“…For example the autotransporter pertactin, other putative adhesins such as BipA and the prominent outer membrane porin protein BP0840 [44][45][46] were significantly upregulated in biofilm. For a number of microorganisms the expression of surface proteins, like BapA in Staphylococcus aureus and Ag43 in Escherichia coli [47,48], has been reported to be essential for proper biofilm formation.…”

Metabolic Pathway for Biosynthesis of Poly (3‐Hydroxybutyrate‐co‐4‐Hydroxybutyrate) from 4‐Hydroxybutyrate by Alcaligenes eutrophus

“…Downstream of ORF2, and in the opposite orientation, ORF9 was identified. The amino acid sequence deduced from the nucleotide sequence of ORF9 exhibited 34.3% amino acid identity to a porin of Bordetella pertussis (Li et al, 1991).…”