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Cloning and expression of the Haemophilus influenzae transferrin receptor genes
Author(s) -
Loosmore Sheena M.,
Yang Yanping,
Coleman Debbie C.,
Shortreed Jean M.,
England Diane M.,
Harkness Robin E.,
Chong Pele S.C.,
Klein Michel H.
Publication year - 1996
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1046/j.1365-2958.1996.406943.x
Subject(s) - biology , cloning (programming) , transferrin , haemophilus influenzae , transferrin receptor , gene , genetics , haemophilus , microbiology and biotechnology , computational biology , receptor , bacteria , biochemistry , computer science , programming language
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.