Cloning structural genes for Treponema pallidum immunogens and characterisation of recombinant treponemal surface protein, P2 (P2 star).
A genomic library consisting of partially digested 10 to 20 kilobase pair fragments of Treponema pallidum deoxyribonucleic acid (DNA) was constructed using bacteriophage lambda EMBL-3 as the vector. Positive clones expressing T pallidum antigens were detected with sera from experimentally infected rabbits. Treponemal proteins ranging in molecular weight from 37,000 daltons to 120,000 daltons were identified by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immunoblotting of phage lysate proteins. One recombinant phage was examined further and contained an insert encoding a prominent treponemal 37,000 dalton protein. The recombinant protein was not recognised by antiserum directed against a fibronectin binding treponemal adhesion that contained the same electrophoretic mobility. Neither did antibody to the recombinant 37,000 dalton protein react with any treponemal proteins purified by fibronectin affinity chromatography. The recombinant protein in Escherichia coli lysates was labelled P2 (P2 star) to differentiate it from the comigrating adhesin protein called P2. Native P2 protein was present on T pallidum surfaces as shown by radioimmunoprecipitation assays with extrinsically labelled organisms. A cross reactive molecule like P2 was not synthesised by the avirulent spirochaete, T phagedenis biotype Reiter, which indicated that P2 is a protein specific to virulent T pallidum organisms. Finally, only sera of patients with primary syphilis possessed appreciable concentrations of antibody to recombinant P2 protein.
Peterson
K M
KM
Department of Microbiology, University of Texas Health Science Center, San Antonio 78284.
Baseman
J B
JB
Alderete
J F
JF
D000907:Antibodies, Bacterial / Q000032:analysis; D001426:Bacterial Proteins / Q000276:immunology / Q000302:isolation & purification*; D003001:Cloning, Molecular / Q000379:methods*; D005796:Genes; D008565:Membrane Proteins / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification*; D013587:Syphilis / Q000276:immunology; D014210:Treponema pallidum / Q000235:genetics*
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins