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| 大肠杆菌邻氨基苯甲酸合成酶编码基因trpED的克隆与表达 | |
| 引用本文： | 李剑欣,郭长江,刘云,林维平,张绪梅,徐琪寿.大肠杆菌邻氨基苯甲酸合成酶编码基因trpED的克隆与表达[J].生物技术通讯,2007,18(2):183-185. |
| 作者姓名： | 李剑欣 郭长江 刘云 林维平 张绪梅 徐琪寿 |
| 作者单位： | 1. 军事医学科学院,放射医学研究所,北京,100850 2. 军事医学科学院,食品与环境卫生研究所,天津,300050 3. 南昌大学,生命科学学院,江西,南昌,330000 |
| 基金项目： | 国家自然科学基金;天津市科技攻关项目 |
| 摘 要： | 目的:通过基因重组的方法获得在体外具有邻氨基苯甲酸合成酶活性的基因trpED,为进一步解除反馈抑制的基因改造寻找靶标。方法:分别构建重组质粒pBV220-trpE和pBV220-trpD;由于trpE和trpD存在翻译偶联现象,将其自大肠杆菌基因组中经PCR扩增得到,构建新的表达质粒pBV220-trpED。结果:SDS-PAGE显示,包含质粒pBV220-trpED的重组菌在相对分子质量约53000(trpE基因表达)和56000(trpD基因表达)处的蛋白表达量明显增多,且邻氨基苯甲酸合成酶活性是对照的3倍。结论:翻译偶联的存在直接影响蛋白活性的发挥;重组质粒pBV220-trpED的获得为进一步的基因改造,以提高色氨酸产量奠定了研究基础。 |
| 关 键 词： | trpED基因 邻氨基苯甲酸合成酶 色氨酸 生物合成 |
| 文章编号： | 1009-0002（2007）02-0183-03 |
| 修稿时间： | 2006-09-13 |
The Clone and Expression of the Anthranilate Synthetase Coding Gene in Escherichia Coli |
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| LI Jian-xin,GUO Chang-jiang,LIU Yun,LIN Wei-ping,ZHANG Xu-mei,XU Qi-shou.The Clone and Expression of the Anthranilate Synthetase Coding Gene in Escherichia Coli[J].Letters in Biotechnology,2007,18(2):183-185. | |
| Authors: | LI Jian-xin GUO Chang-jiang LIU Yun LIN Wei-ping ZHANG Xu-mei XU Qi-shou |
| Institution: | 1. a. Institute of Radiation Medicine, Beijing 100850; b. Institute of Nutration and Sanitation, Tianjin 300050; Academy of Military Medical Sciences; 2. Biology Academy of Nanchang University, Nanchang 330000; China |
| Abstract: | Objective:To get the most active anthranilate synthetase coding gene in vitro,and to build the fundation for the further releasing feed-back inhibition.Methods:Because of the overlapping gene between trpE and trpD,the trpED was cloned integrallty to produce a new high-expression vector pBV220-trpED.Results:The SDS-PAGE indicated that the protein expression of the recombined bacterium had an obvious increase in aim sites,and the enzyme assays showed the recombined one has three times activity of the antitheses.Conclusion:The overlapping phenomenon in Escherichia coli directly influenced the protein activity;and all of these production and data will be elements of the following work to improve the output of tryptophan. |
| Keywords: | trpED gene anthranilate synthetase tryptophan biosynthesis |
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