title: Neisseria meningitidis transferrin-binding protein 1 expressed in Escherichia coli is surface exposed and binds human transferrin.
abstract: A gene library of Neisseria meningitidis B15 P1.16 DNA was established in lambda 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 tbp-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.
authors.affiliation: Department of Microbiology, Queens Medical Centre, University Hospital, Nottingham, UK.
authors.forename: H M
authors.initials: HM
authors.lastname: Palmer
authors.forename: N B
authors.initials: NB
authors.lastname: Powell
authors.forename: D A
authors.initials: DA
authors.lastname: Ala'Aldeen
authors.forename: J
authors.initials: J
authors.lastname: Wilton
authors.forename: S P
authors.initials: SP
authors.lastname: Borriello
mesh_terms: D000595:Amino Acid Sequence; D001419:Bacteria / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism / Q000648:ultrastructure; D015723:Gene Library; D005798:Genes, Bacterial; D006801:Humans; D007501:Iron / Q000378:metabolism; D033862:Iron-Binding Proteins; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013045:Species Specificity; D014168:Transferrin / Q000378:metabolism*; D033882:Transferrin-Binding Protein B; D033863:Transferrin-Binding Proteins
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D033862:Iron-Binding Proteins; D011993:Recombinant Fusion Proteins; D014168:Transferrin; D033882:Transferrin-Binding Protein B; D033863:Transferrin-Binding Proteins; D007501:Iron