Cloning and characterization of the Pseudomonas aeruginosa pbpB gene encoding penicillin-binding protein 3

Clones containing the pbpB gene which encodes penicillin-binding protein (PBP) 3 of Pseudomonas aeruginosa were detected by hybridization by PCR amplification with primers based on the conserved sequences of high- molecular-weight PBPs. The translated amino acid sequence demonstrated 45% identity and had a total of 66% conserved amino acids relative to the Escherichia coli PBP3. The pbpB gene was located upstream of a gene homologous to the E. coli murE gene, which encodes uridine diphosphate-N- acetyl muramic acid-tripeptide synthetase. The overexpressed pbpB gene product reacted with ³H-penicillin G and had an apparent molecular weight of 60,000.