遗传 ›› 2009, Vol. 31 ›› Issue (1): 75-82.doi: 10.3724/SP.J.1005.2009.00075

• 研究报告 • 上一篇    下一篇

Mx基因稀有密码子和mRNA结构及大肠杆菌表达 优化

尹春光1, 2;杜立新1;赵桂平3;李宏滨1

  

  1. 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;
    2. 山东省济宁学院生物系, 济宁 273155;
    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

  • 收稿日期:2008-05-08 修回日期:2008-09-15 出版日期:2009-01-10 发布日期:2009-01-10
  • 通讯作者: 杜立新

Optimizing the expression of Mx gene in Escherichia coli based on rare codon and mRNA structure

YIN Chun-Guang1,2;DU Li-Xin1;ZHAO Gui-Ping3;LI Hong-Bin1   

  1. 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
  • Received:2008-05-08 Revised:2008-09-15 Online:2009-01-10 Published:2009-01-10
  • Contact: DU Li-Xin1

关键词: 稀有密码子, 翻译起始区, Mx蛋白

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.