期刊论文

Expression and Identification of Recombinant Protein Tp1038 of Treponema Pallidum

中文

医学信息

1006-1959

2013

14

266-267

10.3969/j.issn.1006-1959.2013.14.366

湖南省研究生培养创新项目 湖南省自然科学基金重点项目 国家自然科学基金

本校为第一机构

目的表达梅毒螺旋体(Treponema pal idum, Tp)重组蛋白Tp1038(rTp1038)，鉴定其抗原性，为进一步探讨其在梅毒血清学诊断中的价值奠定基础。方法PCR扩增Tp1038全长基因，构建含6个组氨酸(His)标签的原核表达重组体pET-28a(+)/Tp1038，转化宿主菌 E. coli诱导表达 rTp1038，以 Ni-NTA亲和层析柱纯化蛋白，SDS-PAGE分析蛋白表达形式与纯度，Western blot鉴定rTp1038的抗原性。结果成功构建了表达重组体pET-28a/Tp1038，经诱导宿主菌高效表达了一分子量大约为20 KDa的重组蛋白，以可溶性表达形式为主，纯化蛋白的纯度>98%；Western blot结果显示该纯化蛋白仅能被抗 His单抗和梅毒患者混合血清所特异...

Objective To express and identify recombinant protein Tp1038 (rTp1038) of Treponema pal idum (Tp), providing a basis for further investigation of its significance in syphilis serodiagnosis. Methods PCR was used to amplify ful length of Tp1038 gene. The prokaryotic expression recombinant pET-28a (+)/Tp1038 was constructed and transformed into E.coli BL21 for expressing rTp1038. rTp1038 were purified with Ni-NTA af inity chromatography. SDS-PAGE was used to analyse expression form and purity of rTp1038. Western blot was performed to identify antigenicity of rTp1038. Results The prokaryotic recom...