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
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Bacterial Proteins*
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Base Sequence
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Carrier Proteins*
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Cloning, Molecular
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Escherichia coli / genetics
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Gene Expression
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Genes, Bacterial / genetics*
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Hexosyltransferases / genetics*
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Hexosyltransferases / isolation & purification
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Hexosyltransferases / metabolism
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Molecular Sequence Data
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Multienzyme Complexes / genetics*
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Multienzyme Complexes / isolation & purification
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Multienzyme Complexes / metabolism
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Muramoylpentapeptide Carboxypeptidase*
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Neisseria gonorrhoeae / genetics*
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Neisseria meningitidis / genetics*
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Penicillin V / metabolism
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Penicillin-Binding Proteins
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Peptidyl Transferases / genetics*
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Peptidyl Transferases / isolation & purification
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Peptidyl Transferases / metabolism
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Protein Binding
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Recombinant Fusion Proteins / metabolism
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Restriction Mapping
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Sequence Analysis
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Sequence Analysis, DNA
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Sequence Homology, Amino Acid
Substances
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Bacterial Proteins
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Carrier Proteins
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Multienzyme Complexes
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Penicillin-Binding Proteins
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Recombinant Fusion Proteins
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Peptidyl Transferases
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Hexosyltransferases
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Muramoylpentapeptide Carboxypeptidase
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Penicillin V
Associated data
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GENBANK/U72876
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GENBANK/U80933