P.70 pertactin, an outer‐membrane protein from Bordetella parapertussis: cloning, nucleotide sequence and surface expression in Escherichia coli

Abstract

The gene prn encoding the outer-membrane protein P.70 (pertactin) from Bordetella parapertussis has been cloned in Escherichia coli and its DNA sequence determined. Analysis of the DNA sequence reveals that the gene has an open reading frame comprising 922 amino acids capable of encoding a protein with a molecular weight of 95,177 (P.95). In vivo processing of this precursor yields a protein with an estimated Mr of 70 kDa (P.70) which is located on the surface of B. parapertussis. Homology between the prn gene from B. parapertussis and that from Bordetella pertussis is 91.3%. The homology is 93% when the protein sequence of P.95 is aligned with that of P.93 from B. pertussis. The major differences between the P.70 pertactin from B. parapertussis and the P.69 pertactin from B. pertussis occur in the number of reiterated units within the repeat motifs found in both proteins; the sequence Gly-Gly-Xaa-Xaa-Pro is repeated four times in the P.70 pertactin, and five times in the P.69 pertactin, while the sequence Pro-Gln-Pro occurs nine times in P.70 pertactin and five times in P.69 pertactin. Cloning of the gene for P.95 in an E. coli expression vector results in the synthesis of a protein that mimics native gene expression in B. parapertussis, i.e. the P.95 protein is synthesized and subsequently processed to yield the P.70 form of the protein on the surface of the cell.

Novel, host-restricted genotypes of Bordetella bronchiseptica associated with phocine respiratory tract isolates

“…An additional human isolate, 97-219, was reported, paradoxically, as having both the 1-4a1/2-6a1 and the 1-3a/2-6a1 alleles (Diavatopoulos et al, 2005). The region 1 variant 1-4a1 has additionally been reported for 12 human isolates of Bordetella parapertussis, but accompanied only by region 2 sequences comprising eight to ten PQP repeats (Li et al, 1991;Mäkinen et al, 2003;Diavatopoulos et al, 2005;Bouchez et al, 2011). In B. bronchiseptica, 1-4a1 has been found in association with only the six-repeat region 2 variant 2-6a1 Isolate 22/M63/4/09, the sole seal isolate in our study with a genotype found in other hosts (RT1/ST14), was cultured from the intestine of a dead seal recovered on the Atlantic coast of south-west England.…”

Role of the polymorphic region 1 of the Bordetella pertussis protein pertactin in immunity

“…The pertactin genes of isolates used in this study were sequenced previously or in this study (Mooi et al, 1998Mastrantonio et al, 1999 ;Li et al, 1991) (Table 1). Except for the clinical isolates, the prn genes were sequenced completely on both strands.…”

Bi-Antigenic Immunoassay Models Based on the Recombinant PvpA Proteins for Mycoplasma Gallisepticum Diagnosis in Chickens

“…Most likely, the major antigenic domains take place in the C-terminal region containing proline-rich repeat units, known to be highly immunoreactive and involved in the binding of surface-exposed adhesive molecules to host cells. 1,7,13,18 Further investigation is needed to determine the immunodominant fragments of the M. gallisepticum PvpA cytadhesin and to evaluate their adhesive functionality.…”