Hybrid genes over-express pertactin from Bordetella pertussis.
Pertactin is a surface adhesin of Bordetella pertussis which is produced in small quantities when expressed from the native prn promoter. Hybrid genes were constructed in which the prn promoter was replaced by either the fha or tox promoter. Recombinant B. pertussis strains containing chromosomally integrated hybrid tox promoter/prn (toxpprn) or fha promoter/prn (fhapprn) genes expressed pertactin at approximately 5- and 8-fold the wild-type level, respectively. The pertactin was correctly processed and secreted and was biochemically and antigenically comparable to its wild-type counterpart, as determined by N-terminal sequence analysis, immunoblotting, peptide mapping, circular dichroism and antigenicity studies. In an adherence assay, a strain over-expressing pertactin was no more adherent than the wild-type strain, but a pertactin-deficient strain was less adherent.
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D001886:Bordetella pertussis / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D005784:Gene Amplification; D015870:Gene Expression; D005798:Genes, Bacterial; D006168:Guinea Pigs; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins; D011995:Recombination, Genetic; D010566:Virulence Factors, Bordetella
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004274:DNA, Recombinant; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; C068929:pertactin