Serological characterization and gene localization of an Escherichia coli-expressed 37-kilodalton Treponema pallidum antigen.
A recombinant plasmid containing a 5.6-kilobase-pair DNA fragment of the Treponema pallidum genome was characterized by endonuclease mapping, and the encoded proteins were expressed in Escherichia coli and analyzed by use of in vitro transcription and translation. One of the proteins, identified as having a molecular weight of 37,000 (37K protein), was selected for further study. Initially, the seroreactivity of the partially purified 37K antigen was demonstrated by immunoblotting. After its purification to near homogeneity, the cloned T. pallidum protein was assessed for diagnostic significance by radioimmunoassay. Although first identified as seroreactive by screening with secondary syphilitic sera (T. E. Fehniger, A. M. Walfield, T. M. Cunningham, J. D. Radolf, J. N. Miller, and M. A. Lovett, Abstr. Annu. Meet. Am. Soc. Microbiol. 1985, B156, p. 44), the antigen was shown to be serologically reactive with antibodies in serum from all stages of syphilis but was not recognized by serum from controls by both immunoblotting and radioimmune assay. Further, a monospecific polyclonal rabbit antiserum generated to the 37K antigen recognized a polypeptide of the same molecular weight from T. pallidum but did not efficiently recognize proteins from five nonpathogenic treponemes tested. Therefore, because of reactivity with and specificity for T. pallidum antibodies, the 37K antigen may be of serodiagnostic value in the detection of syphilis.
D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D003429:Cross Reactions; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007163:Immunosorbent Techniques; D008970:Molecular Weight; D010957:Plasmids; D011863:Radioimmunoassay; D011994:Recombinant Proteins / Q000276:immunology; D012698:Serologic Tests; D013587:Syphilis / Q000175:diagnosis / Q000276:immunology*; D014210:Treponema pallidum / Q000235:genetics / Q000276:immunology*
D000953:Antigens, Protozoan; D011994:Recombinant Proteins