| 刘小军，牛淑会， 蔡嘉怡，赵飞骏，余坚，姚玲， 吴移谋， 曾铁兵.牛淑会为并列第一作者.梅毒螺旋体重组蛋白Tp0608的表达与鉴定.[J].中南医学科学杂志.,2013,41(4):341-343. |
| 牛淑会为并列第一作者.梅毒螺旋体重组蛋白Tp0608的表达与鉴定 |
| Expression and Identification of Recombinant Protein Tp0608 of Treponema Pallidum |
| 投稿时间：2013-04-01 |
| DOI： |
| 中文关键词: 梅毒 梅毒螺旋体 Tp0608 重组抗原 抗原性 |
| 英文关键词:syphilis Treponema pallidum Tp0608 recombinant antigen antigenicity |
| 基金项目:国家自然科学基金(81273322)；湖南省自然科学基金重点项目(11JJ2044)；湖南省研究生科研创新项目(CX2011B380). |
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| 中文摘要: |
| 目的表达梅毒螺旋体(Tp)重组蛋白Tp0608(rTp0608)并鉴定其抗原性，为深入探讨其在梅毒血清学诊断中的价值奠定基础。方法PCR扩增Tp0608全长基因，构建原核表达重组体pET-28a(+)/Tp0608，转化宿主菌诱导表达rTp0608，Ni-NTA亲和层析法纯化蛋白，SDS-PAGE分析蛋白表达形式与纯度，Western blot鉴定rTp0608的抗原性。结果原核表达重组体pET-28a(+)/Tp0608在宿主菌内经诱导表达了分子量大约为38 kDa的重组蛋白，以包涵体表达形式为主，纯化蛋白的纯度>95%；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PET-28a（+）表达载体高效表达了rTp0608，该重组抗原有良好的抗原性。 |
| 英文摘要: |
| ObjectiveTo express and identify antigenicity of recombinant protein Tp0608 (rTp0608) of Treponema pallidum (Tp),providing a basis for further investigation of its significance in syphilis serodiagnosis.MethodsFull length of Tp0608 gene was amplified by using PCR.The prokaryotic expression recombinant pET-28a(+)/Tp0608 was constructed and transformed into E.coli BL21 to express rTp0608,and rTp0608 was purified with Ni-NTA affinity chromatography.Expression and purity of rTp0608 were analysed by SDS-PAGE.Antigenicity of rTp0608 was tested by Western blot.ResultsThe prokaryotic recombinant pET-28a(+)/Tp0608 was constructed successfully and an approximate 38 KDa recombinant protein was expressed efficiently as inclusion body in E.coli and the protein purity was over 95%.Western blot indicated that the purified proteins were able to be recognized specifically by sera from syphilis patients.Conclusions The efficiently expressed recombinant Tp0608 has good antigenicity. |
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