| 摘要 |
The present disclosure provides the complete primary amino acid, and underlying DNA, sequence for the 47-kilodalton surface immunogen of Treponema pallidum, subsp. pallidum. The sequence was obtained by using a combined strategy of DNA sequencing of the cloned gene as well as confirmatory N-terminal amino acid sequencing of the native antigen. An open reading frame corresponding to the 47-kDa antigen was comprised of 367 amino acid codons, which gave rise to a calculated molecular weight for the corresponding antigen of about 40,701. Also disclosed are methods for preparing variant and mutant molecules having biologically similar attributes, as well as methods for preparing particular antigenic/immunogenic subportions of the 47-kDa protein. In particular aspects, antigenic/immunogenic subportions are identified by hydrophilicity analysis of the protein sequence. The 47-kDa antigen and antigenic subportions of the present invention can be used both as antigens in the detection of clinical materials having anti-47-kDa antibodies therein, as well as in the preparation of vaccines for use in connection with promoting an immune state in vaccinated individuals. Also disclosed are DNA sequences which may be useful both in the preparation of second generation antigens, and as hybridization probes in the detection of pathogenic T. pallidum in clinical samples. Particular methods and embodiments are also disclosed which allow greatly improved recombinant DNA production of the 47-kDa antigen, including placement of the gene under the control of the T7 RNA polymerase promoter.
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