Expression cloning of Pneumocystis carinii antigens.
We undertook expression cloning of Pneumocystis carinii antigens to overcome the difficulties encountered in purification of these antigens. Using monoclonal antibodies to the P. carinii gp120 antigen and polyclonal rabbit antiserum to rat-derived P. carinii, we have isolated cDNA clones encoding immunoreactive moieties. A cDNA clone encoding the 3' portion of a 45-55 kDa antigen of rat-derived P. carinii, was the most abundant clone isolated. The peptide encoded by this cDNA has a novel sequence with a repeated motif rich in glutamic acid residues. Affinity-purified antibodies to this peptide reacted with the 45-55 kDa band of rat-derived P. carinii. The fusion protein was recognized by serum antibodies from rats with natural exposure to P. carinii. The production of this recombinant protein should allow more detailed studies of the host-parasite relationship of this important opportunistic infection.
Smulian
A G
AG
Department of Medicine, University of Cincinnati College of Medicine, Ohio 45267.
Stringer
J R
JR
Linke
M J
MJ
Walzer
P D
PD
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal / Q000235:genetics*; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000302:isolation & purification; D005656:Fungal Proteins / Q000235:genetics / Q000276:immunology; D015723:Gene Library; D008969:Molecular Sequence Data; D011010:Pneumocystis / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis
D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D004271:DNA, Fungal; D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins