梅毒螺旋体重组蛋白Tp0463的表达与抗原性鉴定

目的表达梅毒螺旋体（Tp）重组蛋白Tp0463（rTp0463）并鉴定其抗原性,为进一步探讨其在梅毒血清学诊断和免疫保护中的作用奠定基础。方法 PCR扩增Tp0463基因,构建原核表达重组体pET-28a（＋）/Tp0463,转化宿主菌诱导表达重组蛋白,Ni-NTA亲和层析法纯化蛋白,Westernblot鉴定表达产物的抗原性。结果成功构建了原核表达重组体pET-28a（＋）/Tp0463,经诱导高效表达了一分子量大约为16KDa的重组蛋白,以包涵体表达形式为主,纯化蛋白的纯度〉90%;Westernblot结果显示纯化蛋白能被梅毒患者混合血清特异性识别。结论高效表达了rTp0463,该重组抗原有良好的抗原性。

Expression and antigenicity idenfication of recombinant protein Tp0463 of Treponema pallidum

Objective To express and identify antigenicity of recombinant protein Tp0463（rTp0463） of Treponema pallidum（Tp）,and to provide a basis for further investigation of its significance in syphilis serodiagnosis and immunoprotection.Methods Tp0463 gene was amplified by PCR.The prokaryotic expression recombinant pET-28a（＋）/Tp0463 was constructed and transformed into E.coli BL21 for induced expression of recombinant proteins which were purified with Ni-NTA affinity chromatography.Western blot was performed to identify antigenicity of rTp0463.Results The prokaryotic recombinant pET-28a（＋）/Tp0463 was constructed successfully and an approximate 16KDa recombinant protein was expressed efficiently in E.coli with inclusion body as dominant form and more than 90% purity.Western blot indicated that purified proteins were able to be recognized specially by pooled sera from syphilis patients.Conclusion The recombinant protein Tp0463 expressed efficiently in E.coli has good antigenicity.