期刊论文

Expression and antigenicity idenfication of recombinant protein Tp0463 of Treponema pallidum

中文

中国现代医药杂志

1672-9463

2011

13

9

4-6

10.3969/j.issn.1672-9463.2011.09.002

湖南省科技厅重点项目（编号:2010FJ2008）； 湖南省自然科学基金重点项目（编号:11JJ2044）；

本校为第一机构

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,Western blot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%;Western blot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.

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.Re...