Peterson K M, Baseman J B, Alderete J F
Department of Microbiology, University of Texas Health Science Center, San Antonio 78284.
Genitourin Med. 1987 Oct;63(5):289-96. doi: 10.1136/sti.63.5.289.
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.
以噬菌体λEMBL - 3为载体构建了一个基因组文库，该文库由部分消化的梅毒螺旋体脱氧核糖核酸（DNA）的10至20千碱基对片段组成。用实验感染兔子的血清检测表达梅毒螺旋体抗原的阳性克隆。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳和噬菌体裂解物蛋白的免疫印迹法鉴定了分子量在37,000道尔顿至120,000道尔顿之间的梅毒螺旋体蛋白。进一步检查了一个重组噬菌体，其包含一个编码一种突出的37,000道尔顿梅毒螺旋体蛋白的插入片段。该重组蛋白不被针对具有相同电泳迁移率的纤连蛋白结合梅毒螺旋体黏附素的抗血清识别。针对重组37,000道尔顿蛋白的抗体也不与通过纤连蛋白亲和层析纯化的任何梅毒螺旋体蛋白发生反应。大肠杆菌裂解物中的重组蛋白被标记为P2（P2星号），以将其与称为P2的共迁移黏附素蛋白区分开来。用外部标记的生物体进行放射免疫沉淀分析表明，天然P2蛋白存在于梅毒螺旋体表面。无毒螺旋体T phagedenis生物型Reiter不合成类似P2的交叉反应分子，这表明P2是毒力梅毒螺旋体特有的蛋白质。最后，只有一期梅毒患者的血清含有可观浓度的针对重组P2蛋白的抗体。
