Pap pili as a vector system for surface exposition of an immunoglobulin G-binding domain of protein A of Staphylococcus aureus in Escherichia coli.
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.
Steidler
L
L
Laboratory of Molecular Biology, Gent University, Belgium.
Remaut
E
E
Fiers
W
W
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial; D005822:Genetic Vectors; D007074:Immunoglobulin G / Q000378:metabolism*; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013205:Staphylococcal Protein A / Q000235:genetics* / Q000378:metabolism; D013211:Staphylococcus aureus / Q000235:genetics*
C078802:AtpA protein, E coli; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D007074:Immunoglobulin G; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D036781:Fimbriae Proteins