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目的表达苍白密螺旋体(梅毒螺旋体)黏附蛋白Tp0751,纯化表达产物并进行免疫反应性分析,为探索Tp0751重组蛋白在梅毒致病过程中的作用奠定基础。方法通过生物信息学分析,去除Tp0751信号肽序列,构建原核表达体进行诱导表达;Ni亲合层析柱纯化重组蛋白,Western-印迹检测其免疫反应性。结果成功构建了PET-28a(+)-0751原核表达载体,经表达、纯化后获得了相对分子量约为26×103的融合蛋白,其表达产物占全菌总蛋白的>30%,并且主要可溶性形式存在;Western-印迹检测其能与梅毒患者阳性血清发生特异性反应,具有良好的抗原性。结论重组表达的Tp0751黏附蛋白为可溶性蛋白,且具有良好的免疫反应性,为进一步研究其在Tp0751黏附蛋白在梅毒致病过程中的作用和其生物学功能奠定了基础。
Objective To express the adhesion protein Tp0751 of Treponema pallidum (Treponema pallidum) and to purify the expressed product and analyze its immunoreactivity. This study may provide a basis for exploring the role of Tp0751 in the pathogenesis of syphilis. Methods Tp0751 signal peptide sequence was removed by bioinformatics analysis, and the prokaryotic expression vector was constructed and induced to express. The recombinant protein was purified by Ni affinity chromatography and its immunoreactivity was detected by Western blotting. Results The prokaryotic expression vector PET-28a (+) - 0751 was successfully constructed. After fusion, the fusion protein with the relative molecular weight of 26 × 103 was obtained. The expressed product accounted for> 30% of the total bacterial total protein and the main soluble Form; Western-blot detection of syphilis patients with positive serum-specific reaction has good antigenicity. Conclusion The recombinant Tp0751 adherent protein is a soluble protein and has good immunoreactivity, which lays the foundation for further study on the role of Tp0751 adhesion protein in the pathogenesis of syphilis and its biological functions.