| 弗氏2a志贺氏菌2457T株YciD蛋白的融合表达和纯化 |
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| 引用本文： | 刘先凯,高美琴,孙忠科,冯尔玲,朱力,王恒樑. 弗氏2a志贺氏菌2457T株YciD蛋白的融合表达和纯化[J]. 生物技术通讯, 2009, 20(1): 1-3. DOI: 10.3969/j.issn.1009-0002.2009.01.001 |
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| 作者姓名： | 刘先凯 高美琴 孙忠科 冯尔玲 朱力 王恒樑 |
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| 作者单位： | 1. 军事医学科学院,生物工程研究所,北京,100071
2. 军事医学科学院,生物工程研究所,北京,100071;华中农业大学,食品科技学院,湖北,武汉,430070 |
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| 基金项目： | 国家自然科学基金，国家重点基础研究发展规划(973计划) |
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| 摘 要： | 目的：原核表达重组弗氏2a志贺氏菌2457T株YciD蛋白，为其功能研究奠定基础。方法：用PCR方法从弗氏2a志贺氏菌2457T株染色体中扩增YciD蛋白编码序列，经过纯化、酶切后克隆到原核表达载体pET32a中，构建重组载体pET32a-yciD，转化大肠杆菌BL21（DE3）菌株获得工程菌株，对其表达和纯化条件进行优化；利用Western Blot检测融合蛋白的表达。结果：构建了YciD蛋白的融合表达载体，并在大肠杆菌中获得高效表达；经Ni-NTA亲和层析柱纯化获得了高纯度的YciD蛋白；Western Blot表明，此蛋白可与His标签抗体反应，表明获得了目的蛋白。结论：在原核表达系统中表达、纯化弗氏2a志贺氏菌2457T株YciD蛋白，为进一步对其进行功能研究奠定了基础。
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| 关 键 词： | 弗氏2a志贺氏菌2457T株 YeiD蛋白 融合蛋白 表达 纯化 |
Expression and Purification of the YciD Protein of Shigella flexneri 2a Strain 2457T |
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| LIU Xian-Kai,GAO Mei-Qin,SUN Zhong-Ke,FENG Er-Ling,ZHU Li,WANG Heng-Liang. Expression and Purification of the YciD Protein of Shigella flexneri 2a Strain 2457T[J]. Letters in Biotechnology, 2009, 20(1): 1-3. DOI: 10.3969/j.issn.1009-0002.2009.01.001 |
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| Authors: | LIU Xian-Kai GAO Mei-Qin SUN Zhong-Ke FENG Er-Ling ZHU Li WANG Heng-Liang |
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| Affiliation: | LIU Xian-Kai1,GAO Mei-Qin1,2,SUN Zhong-Ke1,FENG Er-Ling1,ZHU Li1,WANG Heng-Liang11. Beijing Institute of Biotechnology,Beijing 100071,2. Collage of Food Science and Technology,Huazhong Agariculture University,Wuhan 430070,China |
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| Abstract: | Objective: To express and purify YciD protein of Shigella flexneri 2a strain 2457T in vitro. Methods: Thegene fragment coding YciD protein was amplified by PCR and inserted into plasmid pET32a to construct a recombinantvector pET32a-yciD, then the recombinant vector was transformed into E.coli BL21(DE3), and the expression and purifica-tion of recombinant YciD protein was optimized. The fusion protein was characterized by Western blot. Results: ThepET32a-yciD vector was successfully constructed and the reco... |
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| Keywords: | Shigella flexneri 2a strain 2457T YciD protein protein expression protein purification |
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