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| Mx基因稀有密码子和mRNA结构及大肠杆菌表达 优化 | |
| 引用本文： | 尹春光,杜立新,赵桂平,李宏滨.Mx基因稀有密码子和mRNA结构及大肠杆菌表达 优化[J].遗传,2009,31(1):75-82. |
| 作者姓名： | 尹春光 杜立新 赵桂平 李宏滨 |
| 作者单位： | 1. 中国农业科学院北京畜牧兽医研究所国家畜禽分子遗传育种中心,北京,100193;山东省济宁学院生物系,济宁,273155 2. 中国农业科学院北京畜牧兽医研究所国家畜禽分子遗传育种中心,北京,100193 3. 中国农业科学院北京畜牧兽医研究所家禽遗传育种研究室,北京,100193 |
| 基金项目： | 北京市自然科学基金，国家高技术研究发展计划(863计划) |
| 摘 要： | 通过对稀有密码子和mRNA翻译起始区二级结构的分析, 构建了4种重组表达菌株BL21(DE3)/pET-Mx, Rosseta(DE3)/pET-Mx, BL21(DE3)/pGEX-Mx和Rosseta(DE3)/pGEX-Mx, 在大肠杆菌中进行Mx基因的表达, Rosseta(DE3)/pET-Mx和Rosseta(DE3)/pGEX-Mx重组表达菌中都获得了表达, Western blotting检测到了特异的75 kDa表达产物。实验结果证明稀有密码子和mRNA翻译起始区二级结构对Mx 蛋白表达都有影响, 选择适用于稀有密码子表达的菌株Rosetta(DE3)有利于Mx蛋白的表达, 同时翻译起始区二级结构能值较低的表达载体pGEX-Mx获得的表达量明显增高。实验中首次获得了重组表达鸡全长Mx蛋白的大肠杆菌重组菌。 |
| 关 键 词： | Mx蛋白 翻译起始区 稀有密码子 |
| 收稿时间： | 2008-05-08 |
| 修稿时间： | 2008-09-15 |
Optimizing the expression of Mx gene in Escherichia coli based on rare codon and mRNA structure |
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| YIN Chun-Guang,DU Li-Xin,ZHAO Gui-Ping,LI Hong-Bin.Optimizing the expression of Mx gene in Escherichia coli based on rare codon and mRNA structure[J].Hereditas,2009,31(1):75-82. | |
| Authors: | YIN Chun-Guang DU Li-Xin ZHAO Gui-Ping LI Hong-Bin |
| Institution: | 1. National Center for Molecular Genetics and Breeding of Animal, Institute of BeiJing Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China; 2. Biology Departmeng of Jining University, Shandong 273155, China; 3. Poultry Genetic and Breeding Department, Institute of Beijing Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China |
| Abstract: | Rare codon and mRNA secondary structure of translation initiation region were analyzed. Four bioengineered bacterium BL21(DE3)/pET-Mx, Rosseta (DE3)/pET-Mx, BL21(DE3)/pGEX-Mx, and Rosseta (DE3)/pGEX-Mx were obtained. Through optimizing the rare codon and mRNA secondary structure of translation initiation region, Mx protein was expressed in Rosseta (DE3)/pET-Mx, and Rosseta (DE3)/pGEX-Mx. The specified product of 75 kDa was detected by means of Western blotting analysis. The result showed that Rosetta (DE3) strain expressing some rare codon used in our experiment can obtain Mx protein expression, and the lower energy of mRNA structure can improve the expression level of Mx protein. This is the first report on the expression of the full open reading frame of Mx gene. |
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