梅毒螺旋体重组蛋白Tp0965的表达与鉴定

Expression and Identification of Recombinant Protein Tp0965 of Treponema Pallidum

目的:研究梅毒螺旋体(Tp)重组蛋白Tp0965(r Tp0965)的表达并鉴定其免疫反应性,为进一步探讨其在梅毒血清学诊断与疫苗中的应用价值奠定基础。方法:构建Tp0965原核表达重组体p ET-28a(+)/Tp0965,诱导转化宿主菌表达重组蛋白,Ni-NTA亲和层析法纯化蛋白,Western blot鉴定其免疫反应性。结果:成功构建了原核表达重组体p ET-28a(+)/Tp0965,经诱导高效表达了一分子量约为43 KDa的可溶性重组蛋白,纯化蛋白纯度>95%;Western blot显示该纯化蛋白能被梅毒患者血清特异性识别。结论:高效表达了可溶性r Tp0965,该重组抗原有良好的免疫反应性。

Objective：To express and identify immunoreactivity of recombinant Tp0965（r Tp0965） protein of Treponema pallidum（Tp）, provid a basis for further investigation of its significance in syphilis serodiagnosis and vaccine.Method：The prokaryotic expression recombinant p ET-28a（＋）/Tp0965 was constructed and induced to express recombinant proteins which then were purified with Ni-NTA affinity chromatography. Western blot was used to identify immunoreactivity of r Tp0965.Result：The recombinant p ET-28a（＋）/Tp0965 was constructed successfully and a soluble recombinant protein with approximate 43 KDa molecular weight was expressed efficiently in E.coli with more than 95% of purity. Western blot indicated that purified proteins were able to be recognized specially by pooled sera from syphilis patients.Conclusion：The recombinant protein Tp0965 is expressed efficiently in E.coli and has good immunoreactivity.