结核分枝杆菌阿拉伯糖基转移酶AftA的原核表达及鉴定.PDF

中国人兽共患病学报 400 2008．24(5) ChineseournalofZoonoses J 文章编号：1002—2694(2008)05—0400—04 结核分枝杆菌阿拉伯糖基转移酶 AftA的原核表达及鉴定 王爽，朱道银，帖儒修，李娜，王渝伟 摘 要：目的 构建离效表达菌株．以大量表达结核分枝杆菌AftA蛋白。为后续的抗结核药物的筛选奠定基础．方法 产物．结果 生特异性反应．结论 成功制备出重组的A以A蛋白，为新型抗结核药物的筛选奠定了物质基础。 关键词：结核分枝杆菌l阿拉伯糖基转移酶IaftA基因；原核表达 中图分类号：R378，91文献标识码：A andidentificationof tuberculosis Expression Mycobacterium protein WANG Ru-xiu，LINa，WANGYu-wei Dao—yin，TIE Shuang，ZHU Medical 400016．China) (Departmentofimmunology。ChongqingUniversity·Chongqing the tO for ABSTRACT矗ocloneand arabinofuranosyltransferase(AftA)proteinprovideactive a8a express proteintarget new was PCRfromgenomeof tuberculosisH37Rvstrainandin- drugdevelopment。theariAgene amplifiedby Mycobacterium serted vector tOobtainrecombinant recombinantvector into expressionpQE30 plasmidpQE30一afta．The pQE30- prokaryotie restrictionendonucleaseand wastrarmfonnedintoE．coliafter by sequenceanalysis．TheE．coliM15trans- aftA digestion ／ormedwith wasinducedwithIPTG，andthe was SDSandWestern-blot— pQE30-aftA expressionproductanalyzedby Ni-NTA wasfoundthat the was by purificationsystem．It therecombinanthada ring．Thenexpressedproteinpurified protein relativemolecul