Cloning and characterization of the ponA gene encoding penicillin-binding protein 1 from Neisseria gonorrhoeae and Neisseria meningitidis

J Bacteriol. 1997 Apr;179(8):2783-7. doi: 10.1128/jb.179.8.2783-2787.1997.

Abstract

The ponA gene encoding penicillin-binding protein 1 (PBP 1) from Neisseria gonorrhoeae was cloned by a reverse genetic approach. PBP 1 was purified from solubilized membranes of penicillin-susceptible strain FA19 by covalent ampicillin affinity chromatography and used to obtain an NH2-terminal amino acid sequence. A degenerate oligonucleotide based on this protein sequence and a highly degenerate oligonucleotide based on a conserved amino acid motif found in all class A high-molecular-mass PBPs were used to isolate the PBP 1 gene (ponA). The ponA gene encodes a protein containing all of the conserved sequence motifs found in class A PBPs, and expression of the gene in Escherichia coli resulted in the appearance of a new PBP that comigrated with PBP 1 purified from N. gonorrhoeae. A comparison of the gonococcal ponA gene to its homolog isolated from Neisseria meningitidis revealed a high degree of identity between the two gene products, with the greatest variability found at the carboxy terminus of the two deduced PBP 1 protein sequences.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins*
  • Base Sequence
  • Carrier Proteins*
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Gene Expression
  • Genes, Bacterial / genetics*
  • Hexosyltransferases / genetics*
  • Hexosyltransferases / isolation & purification
  • Hexosyltransferases / metabolism
  • Molecular Sequence Data
  • Multienzyme Complexes / genetics*
  • Multienzyme Complexes / isolation & purification
  • Multienzyme Complexes / metabolism
  • Muramoylpentapeptide Carboxypeptidase*
  • Neisseria gonorrhoeae / genetics*
  • Neisseria meningitidis / genetics*
  • Penicillin V / metabolism
  • Penicillin-Binding Proteins
  • Peptidyl Transferases / genetics*
  • Peptidyl Transferases / isolation & purification
  • Peptidyl Transferases / metabolism
  • Protein Binding
  • Recombinant Fusion Proteins / metabolism
  • Restriction Mapping
  • Sequence Analysis
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid

Substances

  • Bacterial Proteins
  • Carrier Proteins
  • Multienzyme Complexes
  • Penicillin-Binding Proteins
  • Recombinant Fusion Proteins
  • Peptidyl Transferases
  • Hexosyltransferases
  • Muramoylpentapeptide Carboxypeptidase
  • Penicillin V

Associated data

  • GENBANK/U72876
  • GENBANK/U80933