Pap pili as a vector system for surface exposition of an immunoglobulin G-binding domain of protein A of Staphylococcus aureus in Escherichia coli

Abstract

Fusion genes between papA, the gene coding for the major Pap pilus subunit, and fragments coding for an immunoglobulin G-binding domain of the Staphylococcus aureus protein A were constructed in such a way that the spa fragments were inserted following either codon 7 or 68 of the coding sequence for the mature portion of PapA. Peptides in the area of amino acids 7 and 68 of PapA are localized at the external side of the pilus. A set of pL expression plasmids containing papA and derivatives suitable for insertion were constructed. A papA gene carrying a spa insert following codon 68 was cloned back into the pap operon. The presence of this altered operon in a bacterial strain allowed the detection of immunoglobulin G-binding activity at the surfaces of the bacterial cells.