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齿垢密螺旋体抗原基因tdpA的分子克隆及序列分析

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

作者信息

Miyamoto M, Noji S, Kokeguchi S, Kato K, Kurihara H, Murayama Y, Taniguchi S

机构信息

Department of Periodontology and Endodontology, Okayama University Dental School, Japan.

出版信息

Infect Immun. 1991 Jun;59(6):1941-7. doi: 10.1128/iai.59.6.1941-1947.1991.

Abstract

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.

摘要

我们分离并鉴定了一种口腔螺旋体——齿垢密螺旋体约翰逊株的免疫原性蛋白。以λ噬菌体EMBL3为载体构建的基因组DNA文库,用兔抗全细胞血清进行免疫筛选。通过蛋白质免疫印迹分析,我们发现了一个编码分子量为53,000抗原的特定克隆;我们将该抗原命名为齿垢密螺旋体蛋白A(TdpA)。完整序列测定揭示了一个1419 bp的开放阅读框和一个与细菌脂蛋白信号肽序列同源的信号肽序列。Southern杂交分析表明,tdpA基因在六个测试的齿垢密螺旋体菌株中高度保守。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,尽管抗体的反应性在个体之间有所不同。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3670/257947/c28af803d623/iai00042-0081-a.jpg

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