Molecular analysis of a haemagglutinin of Haemophilus paragallinarum

Microbiology (Reading). 2002 Jul;148(Pt 7):2171-2179. doi: 10.1099/00221287-148-7-2171.

Abstract

The gene encoding a haemagglutinin of H. paragallinarum, hagA, has been identified and the full-length nucleotide sequence determined. A approximately 39 kDa protein, recognized by an anti-haemagglutinin monoclonal antibody, mAb4D, was purified from H. paragallinarum strain 0083 and the N-terminal sequence obtained. The full-length nucleotide sequence was obtained by inverse PCR and the deduced amino acid sequence of the protein encoded was shown to be similar to other outer-membrane proteins of closely related organisms in the HAP group (Haemophilus, Actinobacillus, Pasteurella), especially the P5 protein of Haemophilus influenzae. The hagA gene was cloned into a His-tag expression vector and overexpressed in Escherichia coli strain M15(pREP4). The identity of the purified recombinant protein as a H. paragallinarum haemagglutinin was confirmed by haemagglutination of chicken red blood cells and reactivity, in a Western blot, with the monoclonal antibody specific for the serovar A haemagglutinin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bacterial Proteins*
  • Chickens / microbiology*
  • Cloning, Molecular
  • Haemophilus / classification
  • Haemophilus / genetics
  • Haemophilus / metabolism*
  • Hemagglutination Inhibition Tests
  • Hemagglutination Tests
  • Hemagglutinins / chemistry
  • Hemagglutinins / genetics*
  • Hemagglutinins / isolation & purification
  • Hemagglutinins / metabolism
  • Immunoblotting
  • Lectins
  • Molecular Sequence Data
  • Recombinant Proteins
  • Sequence Alignment
  • Sequence Analysis, DNA

Substances

  • Bacterial Proteins
  • Hemagglutinins
  • Lectins
  • Recombinant Proteins
  • hemagglutinin A, Porphyromonas gingivalis

Associated data

  • GENBANK/AF491817
  • GENBANK/AF491818
  • GENBANK/AF491819
  • GENBANK/AF491820
  • GENBANK/AF491821
  • GENBANK/AF491822
  • GENBANK/AF491823
  • GENBANK/AF491824
  • GENBANK/AF491825
  • GENBANK/AF491826
  • GENBANK/AF491827