Mx基因稀有密码子和mRNA结构及其大肠杆菌表达 优化.pdfVIP

HEREDITAS (Beijing) 2009 1 , 31(1): 75―82 ISSN 0253-9772 研究报告 DOI: 10.3724/SP.J.1005.2009.00075 Mx 基因稀有密码子和mRNA 结构及大肠杆菌表达 优化 1, 2 1 3 1 尹春光 , 杜立新 , 赵桂平 , 李宏滨 1. , 100193; 2. , 273155; 3. , 100193 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。 Mx 蛋白; 翻译起始区; 稀有密码子 Optimizing the expression of Mx gene in Escherichia coli based on rare codon and mRNA structure 1,2 1 3 1 YIN Chun-Guang , DU Li-Xin , ZHAO Gui-Ping , LI Hong-Bin 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 ob- tained. Through optimizing the rare codon and mRNA secondary structure of translation initiation region, Mx protein was expressed i