Open AccessNeisseria meningitidis transferrin‐binding protein 1 expressed in Escherichia coli is surface exposed and binds human transferrin
Author(s) -
Palmer Helen M.,
Powell Nicholas B.L.,
Ala'Aldeen Dlawer A.,
Wilton Jane,
Peter Borriello S.
Publication year - 1993
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1993.tb06310.x
Subject(s) - microbiology and biotechnology , biology , transferrin , gene , dna , gene product , primer (cosmetics) , biochemistry , chemistry , gene expression , organic chemistry
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.