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梅毒螺旋体重组蛋白Tp0965的表达与鉴定 被引量:2
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作者 周秀萍 杨长顺 +4 位作者 牛淑会 李树平 阳大庆 付祥 曾铁兵 《中国医学创新》 CAS 2014年第33期15-17,共3页
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... 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。 展开更多
关键词 梅毒 梅毒螺旋体 tp0965 重组抗原
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Seroreactivity and immunogenicity of Tp0965, a hypothetical membrane protein of Treponema pallidum 被引量:7
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作者 LONG Fu-quan ZHANG Jin-ping +3 位作者 SHANG Guang-dong SHANG Shu-xian GONG Kuang-long WANG Qian-qiu 《Chinese Medical Journal》 SCIE CAS CSCD 2012年第11期1920-1924,共5页
Background Treponema pallidum (T. pallidum) subsp, pallidum is the causative agent of syphilis. Analysis of recombinant antigens of T. pallidum led to the identification of potential candidate antigens for vaccine d... Background Treponema pallidum (T. pallidum) subsp, pallidum is the causative agent of syphilis. Analysis of recombinant antigens of T. pallidum led to the identification of potential candidate antigens for vaccine development and syphilis serodiagnosis. Tp0965 was predicted to be a membrane fusion protein and was found to be reactive with infected human sera in previous studies, but the results were controversial. In this research, the antigenicity and immunoreactivity of recombinant protein Tp0965 were assessed. Methods T. pallidum subsp, pallidum (Nichols strain) was propagated and isolated and the genomic DNA was extracted. The Tp0965 gene was amplified by polymerase chain reaction (PCR). Then the recombinant protein Tp0965 was expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid (Ni-NTA) purification system. The reactivities of protein Tp0965 were examined by immunoblot analysis and indirect enzyme-linked immunosorbent assay. The antisera against protein Tp0965 were obtained by immune rabbits and the immunogenicity of antisera were detected by indirect enzyme-linked immunosorbent assay. Results Recombinant protein Tp0965 was expressed successfully in vitro. Immunoblot assay showed that the recombinant protein Tp0965 could be recognized by human syphilitic sera of all stages. Indirect enzyme-linked immunosorbent assay showed there were only 4 of 74 human syphilitic sera that failed to show reactivity to recombinant antigen Tp0965, and lack of reactivity of Tp0965 to all 28 uninfected sera. A low titer of antiserum against Tp0965 in immune rabbits could be detected after the third time of immunization. Conclusions The recombinant antigen Tp0965 shows excellent sensitivity for the reactivity with sera from syphilitic individuals at all stages. The results also demonstrate a potential application for the serodiagnosis of syphilis. 展开更多
关键词 Treponema pallidum membrane protein recombination reactivity diagnosis tp0965
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