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| 新型佐剂——鞭毛蛋白及其突变体的原核表达和纯化 | |
| 引用本文： | 吴晓芳,李树龙,戴红梅,付桂明,方宏清.新型佐剂——鞭毛蛋白及其突变体的原核表达和纯化[J].生物技术通讯,2011,22(5):691-695. |
| 作者姓名： | 吴晓芳 李树龙 戴红梅 付桂明 方宏清 |
| 作者单位： | 1. 南昌大学食品科学与技术国家重点实验室;南昌大学生命科学与食品工程学院,江西南昌330047;军事医学科学院生物工程研究所,北京100850 2. 军事医学科学院生物工程研究所,北京,100850 3. 南昌大学食品科学与技术国家重点实验室;南昌大学生命科学与食品工程学院,江西南昌330047 |
| 摘 要： | 目的:改构鞭毛蛋白可变区,并优化目的蛋白表达与纯化参数,为评价并获得新型高效蛋白佐剂奠定基础。方法:以鼠伤寒沙门菌基因组为模板钓取2种鞭毛蛋白FljB和FliC的编码基因,分别构建FljB和FliC及其对应的删除可变区的突变体BNLC和CNLC的表达载体,在大肠杆菌中诱导表达,并纯化目的蛋白。结果与结论:原初鞭毛蛋白FljB和FliC均以胞内可溶形式表达,而改造后的鞭毛蛋白BNLC和CNLC则以包涵体形式表达。通过优化纯化条件,分别建立了针对4种蛋白的纯化方法,得到无热源、纯度大于95%的目的蛋白,为进一步研究鞭毛蛋白的结构与佐剂效应的关系奠定了基础。 |
| 关 键 词： | 鞭毛蛋白 突变体 原核表达 纯化 |
Prokaryotic Expression and Purification of Flagellins and Their Mutants as the Neotype Adjuvants |
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| WU Xiao-Fang,LI Shu-Long,DAI Hong-Mei,FU Gui-Ming,FANG Hong-Qing.Prokaryotic Expression and Purification of Flagellins and Their Mutants as the Neotype Adjuvants[J].Letters in Biotechnology,2011,22(5):691-695. | |
| Authors: | WU Xiao-Fang LI Shu-Long DAI Hong-Mei FU Gui-Ming FANG Hong-Qing |
| Institution: | WU Xiao-Fang 1,2,3,LI Shu-Long 3,DAI Hong-Mei 3,FU Gui-Ming 1,FANG Hong-Qing 3 1.State Key Laboratory of Food Science and Technology,Nanchang University,Nanchang 330047,2.Life Science and Food Engineering College,3.Beijing Institute of Biotechnology,Bei-jing 100850,China |
| Abstract: | Objective: This study for deleting of flagellin's central hypervariable domain,optimizating parameters of their protein's expression and purification was carried out,and it should contribute to gaining new and high effi-cient adjuvants actively.Methods: Have fished the genes of two flagellins FljB and FliC from genome of Salmonel-la typhimurium,we constructed the expression vectors for FljB and FliC and their relative mutants BNLC and CNLC,which variable regions were deleted selectively,then we induced the ... |
| Keywords: | flagellin mutants prokaryotic expression protein purification |
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