Cloning and expression of the Haemophilus influenzae transferrin receptor genes

• Published in: Molecular microbiology Vol. 19; no. 3; p. 575
• Main Authors: Loosmore, S M, Yang, Y P, Coleman, D C, Shortreed, J M, England, D M, Harkness, R E, Chong, P S, Klein, M H
• Format: Journal Article
• Language: English
• Published: England 01-02-1996
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Blotting, Western; Carrier Proteins - chemistry; Carrier Proteins - genetics; Carrier Proteins - immunology; Cloning, Molecular; Conserved Sequence; Cross Reactions; Escherichia coli - genetics; Genes, Bacterial; Haemophilus Infections - immunology; Haemophilus Infections - metabolism; Haemophilus influenzae - chemistry; Haemophilus influenzae - genetics; Immune Sera - immunology; Immunization, Passive; Iron-Binding Proteins; Molecular Sequence Data; Plasmids; Rats; Receptors, Transferrin - chemistry; Receptors, Transferrin - genetics; Receptors, Transferrin - immunology; Recombinant Proteins - chemistry; Restriction Mapping; Sequence Homology, Amino Acid; Transferrin-Binding Proteins
• ISSN: 0950-382X
• DOI: 10.1046/j.1365-2958.1996.406943.x

The genomic transferrin receptor genes (tbpA and tbpB) from two strains of Haemophilus influenzae type b (Hib) and two strains of non-typable H. influenzae (NTHi) have been cloned and sequenced. The deduced protein sequences of the H. influenzae tbpA genes were 95-100% conserved and those of the tbpB genes were 66-100% conserved. The tbpB gene from one strain of NTHi was found to encode a truncated Tbp2 protein. The tbpB genes from four additional NTHi strains were amplified by the polymerase chain reaction (PCR) utilizing primers derived from the conserved N-terminal sequences of Tbp1 and Tbp2 and were found to encode full-length proteins. Although several bacterial species express transferrin receptors, when the Tbp1 and Tbp2 sequences from different organisms were compared, there was only limited homology. Recombinant Tbp1 and Tbp2 proteins were expressed from Escherichia coli and antisera were raised to the purified proteins. There was significant antigenic conservation of both Tbp1 and Tbp2 amongst H. influenzae strains, as determined by Western blot analysis. In a passive model of bacteraemia, infant rats were protected from challenge with Hib after transfer of anti-rTbp2 antiserum, but not after anti-rTbp1 antiserum.