| 结核杆菌DnaA蛋白的原核表达及免疫血清的制备和检测 |
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| 引用本文： | 周爱萍,韩蕾,王娟,段艳,吴晓康,徐纪茹.结核杆菌DnaA蛋白的原核表达及免疫血清的制备和检测[J].西安交通大学学报(医学版),2011,32(1):50-53,88. |
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| 作者姓名： | 周爱萍 韩蕾 王娟 段艳 吴晓康 徐纪茹 |
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| 作者单位： | 1. 西安交通大学医学院免疫学与病原生物学系,陕西西安,710061
2. 西安市结核病院,陕西西安,710067 |
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| 基金项目： | 陕西省卫生厅科学研究基金项目 |
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| 摘 要： | 目的 在大肠杆菌中表达重组结核杆菌DnaA蛋白,对表达产物进行纯化、鉴定后制备DnaA蛋白特异性免疫血清并进行Western blot检测.方法 以结核分枝杆菌标准菌株H37Rv基因组为模板,PCR扩增DnaA基因,构建DnaA原核表达质粒pET-DnaA,转化表达宿主大肠杆菌BL21(DE3),IPTG诱导重组蛋白表...
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| 关 键 词： | 结核分枝杆菌 DnaA蛋白 DNA复制起始 原核表达 |
Prokaryotic expression of Mycobacterial Tuberculosis DnaA as well as the preparation and identification of immune serum against DnaA protein |
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| ZHOU Ai-ping,HAN Lei,WANG Juan,DUAN Yan,WU Xiao-kang,XU Ji-ru.Prokaryotic expression of Mycobacterial Tuberculosis DnaA as well as the preparation and identification of immune serum against DnaA protein[J].Journal of Xi‘an Jiaotong University:Medical Sciences,2011,32(1):50-53,88. |
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| Authors: | ZHOU Ai-ping HAN Lei WANG Juan DUAN Yan WU Xiao-kang XU Ji-ru |
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| Institution: | ZHOU Ai-ping1,HAN Lei1,WANG Juan1,DUAN Yan2,WU Xiao-kang1,XU Ji-ru1(1.Department of Immunology and Pathogenic Biology,Medical School ofXi'an Jiaotong University,Xi'an 710061,2.Xi'an Tuberculosis Hospital,Xi'an 710067,China) |
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| Abstract: | Objective To express the recombinant Mycobacterium tuberculosis(Mtb) DnaA protein in E.coli and obtain the specific immune serum against it by purifying and identifying of the expressed product.Methods The DnaA gene of Mtb was amplified by PCR from the Mtb genome and cloned into the expression vector pET-22a(+) to generate the pET-DnaA.The recombinant protein was induced by IPTG and purified via Ni-NTA affinity chromatography.Rabbit was immunized with preliminary purified protein.The immune serum was collec... |
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| Keywords: | Mycobacterium tuberculosis(Mtb) DnaA protein DNA replication prokaryotic expression |
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