Molecular analysis of a haemagglutinin of Haemophilus paragallinarum.
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.
Hobb
Rhonda I
RI
Department of Microbiology and Parasitology, School of Molecular and Microbial Sciences, University of Queensland, St Lucia, QLD 4072, Australia1.
Tseng
Hsing-Ju
HJ
Department of Microbiology and Parasitology, School of Molecular and Microbial Sciences, University of Queensland, St Lucia, QLD 4072, Australia1.
Downes
John E
JE
Department of Microbiology and Parasitology, School of Molecular and Microbial Sciences, University of Queensland, St Lucia, QLD 4072, Australia1.
Terry
Tamsin D
TD
Department of Microbiology and Parasitology, School of Molecular and Microbial Sciences, University of Queensland, St Lucia, QLD 4072, Australia1.
Blackall
Patrick J
PJ
Agency for Food and Fibre Sciences, Animal Research Institute, Department of Primary Industries, Yeerongpilly, Australia2.
Takagi
Masami
M
National Veterinary Assay Laboratory, Dobutsu Iyakuhin Kensajo, 1-15-1 Tokura, Kokubunji-shi, Tokyo, Japan3.
Jennings
Michael P
MP
Department of Microbiology and Parasitology, School of Molecular and Microbial Sciences, University of Queensland, St Lucia, QLD 4072, Australia1.
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins; D002645:Chickens / Q000382:microbiology*; D003001:Cloning, Molecular; D006190:Haemophilus / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D006385:Hemagglutination Inhibition Tests; D006386:Hemagglutination Tests; D006388:Hemagglutinins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015151:Immunoblotting; D037102:Lectins; D008969:Molecular Sequence Data; D011994:Recombinant Proteins; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins; D006388:Hemagglutinins; D037102:Lectins; D011994:Recombinant Proteins; C114923:hemagglutinin A, Porphyromonas gingivalis