Neisseria meningitidis transferrin-binding protein 1 expressed in Escherichia coli is surface exposed and binds human transferrin |
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Authors: | Helen M. Palmer Nicholas B.L. Powell Dlawer A. Ala'Aldeen Jane Wilton S. Peter Borriello |
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Affiliation: | Microbial Pathogenicity Research Group, Clinical Research Initiative in Bacterial Infections, Department of Microbiology, Queens Medical Centre, University Hospital, Nottingham, UK |
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Abstract: | Abstract A gene library of Neisseria meningitidis B15 P1.16 DNA was established in λ Zap II and clones containing DNA encoding transferrin binding protein 1 (TBP-1) identified following hybridisation with a 63-bp DNA probe based on the codon assignment for the first 21 N-terminal amino acids of TBP-1. Sequencing of the cloned DNA demonstrated that all of the intergenic DNA (i.e. upstream of bp-1 running through to the 3' end of the transferrin-binding protein 2 gene) and approx. 15% of tbp-1 had been cloned. The complete gene was generated using a polymerase chain reaction, with the primer for the 3' end being based on tbp-A of N. gonorrhoeae , and the approx. 2.9-kb DNA product cloned into pGem-3Z. The expressed protein (approx. 100 kDa) reacted with antiserum to an N-terminal peptide of TBP-1. In addition, the native product was surface-expressed by Escherichia coli and bound human transferrin. |
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Keywords: | Neisseria meningitidis Transferrin Polymerase chain reaction Transferrin binding protein |
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