Expression and characterization of the ponA (ORF I) gene of Haemophilus influenzae: functional complementation in a heterologous system

A 3.5-kb EcoRI-SphI fragment encompassing the entire wild type ponA gene of haemophilus influenzae is obtained from plasmid pBS98 and cloned into the EcoRI and SphI sites of pMAK705 to obtain pARC0583, which is subsequently used to construct E. coli ARC401. The expression vectors or penicillin-binding proteins (PBPs) of the ARC401 specimen is then studied for their in vivo enzymatic activities. Results indicate that the PBPs possess transglycosylase activity and is capable of maintaining the viability of the E.coli cells in transcomplementations activities.