|
|||||
|
|
| 结核分枝杆菌ESAT-6基因在毕赤酵母中的构建及其表达 | |
| 引用本文： | 付小强,刘洪波,吴少庭.结核分枝杆菌ESAT-6基因在毕赤酵母中的构建及其表达[J].中国热带医学,2011,11(9):1048-1050. |
| 作者姓名： | 付小强 刘洪波 吴少庭 |
| 作者单位： | 1. 鄂州市疾病预防控制中心,湖北鄂州,436000 2. 中山市疾病预防控制中心,广东中山,528400 3. 深圳市疾病预防控制中心,广东深圳,518020 |
| 摘 要： | 目的在毕赤酵母（Pichia pastoris）表达系统中表达结核分枝杆菌ESAT-6基因。方法以重组质粒pET23a-ESAT-6为模板,亚克隆目的片段ESAT-6至酵母菌分泌表达载体pPICZaA,重组质粒经线性化后电转化转染至毕赤酵母菌GS115菌株,经抗生素Zeocin筛选得到高拷贝转化子。经甲醇诱导表达,取细胞裂解上清进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质印迹法分析。结果成功构建了pPICZaA-ESAT-6毕赤酵母表达重组质粒。经甲醇诱导,在酵母细胞内表达分子质量单位为18kDa的蛋白。蛋白质印迹（Western blot）显示,18kDa蛋白被活动性肺结核病人血清抗体识别。结论在毕赤酵母菌中成功表达了带有信号肽ESAT-6基因,为进一步研究新型单位疫苗打下坚实的基础。 |
| 关 键 词： | 结核分枝杆菌 ESAT-6 毕赤酵母菌 基因表达 |
Expression and immunogenicity of the ESAT-6 gene of Mycobaterium tuberculosis in Pichia pastoris |
|
| FU Xiao-qiang,LIU Hong-bo,WU Shao-ting.Expression and immunogenicity of the ESAT-6 gene of Mycobaterium tuberculosis in Pichia pastoris[J].China Tropical Medicine,2011,11(9):1048-1050. | |
| Authors: | FU Xiao-qiang LIU Hong-bo WU Shao-ting |
| Institution: | FU Xiao-qiang,LIU Hong-bo,WU Shao-ting.(1.Ezhou Municipal Centre for Disease Control and Prevention,Ezhou 436000,Hubei,P.R.China,Corresponding author:WU Shao-ting,E-mail:397319588@qq.com) |
| Abstract: | Objective To construct and express the recombinant plasmid pPICZaA-ESAT-6 of Mycobacterium tuberculosis(M.tuberculosis) in Pichia pastoris(P.pastors).Methods ESAT-6 gene was amplified by PCR and cloned into the expression vector pPICZaA of P.pastoris to construct pPICZaA-ESAT-6.After the recombinant was isolated and linearized,it was transformed into P.pastoris GS115 by electro transformation and screened for Zeocin resistance and phenotyping.The high resistant Mut clones were cloned and the lysate of cells... |
| Keywords: | Mycobaterium tuberculosis ESAT-6 genes Pichia pastoris Gene expression Immunogenicity |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |