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Molecular cloning and sequence analysis of antigen gene tdpA of Treponema denticola.

机译:密螺旋体抗原基因tdpA的分子克隆和序列分析。

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摘要

We isolated and characterized an immunogenic protein of an oral spirochete, Treponema denticola Johnson. A genomic DNA library constructed with bacteriophage lambda EMBL3 as a vector was immunologically screened with a rabbit antiserum against the whole cells. Using Western immunoblot analysis, we found a particular clone encoding an antigen with a molecular weight of 53,000; we designated the antigen as T. denticola protein A (TdpA). Complete sequence determination revealed an open reading frame of 1,419 bp and a signal peptide sequence that was homologous to that of bacterial lipoprotein. Southern hybridization analysis revealed that the tdpA gene is highly conserved in six tested strains of T. denticola species. Furthermore, we found that sera from some periodontitis patients contained antibody against the TdpA protein, although the reactivities of the antibodies varied among individuals.
机译:我们分离并表征了一种口服螺旋体,螺旋体密螺旋体约翰逊的免疫原性蛋白。用兔噬菌体EMBL3作为载体构建的基因组DNA文库用针对全细胞的兔抗血清进行免疫筛选。使用Western免疫印迹分析,我们发现了一个特定的克隆,该克隆编码分子量为53,000的抗原。我们将抗原命名为T. denticola蛋白A(TdpA)。完整的序列确定揭示了一个1,419 bp的开放阅读框和一个与细菌脂蛋白同源的信号肽序列。 Southern杂交分析表明,tdpA基因在6个测试的T. denticola种菌株中高度保守。此外,我们发现一些牙周炎患者的血清中含有针对TdpA蛋白的抗体,尽管抗体的反应性因人而异。

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