NEISSERIA-MENINGITIDIS TRANSFERRIN-BINDING PROTEIN-1 EXPRESSED IN ESCHERICHIA-COLI IS SURFACE-EXPOSED AND BINDS HUMAN TRANSFERRIN

A gene library of Neisseria meningitidis B15 P1.16 DNA was established in lambdaZap II and clones containing DNA encoding transferrin bindin g protein 1 (TBP-1) identified following hybridisation with a 63-bp DN A probe based on the codon assignment for the first 21 N-terminal amin o acids of TBP-1. Sequencing of the cloned DNA demonstrated that all o f 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 o f 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 su rface-expressed by Escherichia coli and bound human transferrin.