[Optimizing the expression of Mx gene in Escherichia coli based on rare codon and mRNA structure].
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.
Yin
Chun-Guang
CG
National Center for Molecular Genetics and Breeding of Animal, Institute of BeiJing Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China. sdjnycg@yahoo.com.cn
DU
Li-Xin
LX
Zhao
Gui-Ping
GP
Li
Hong-Bin
HB
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D003062:Codon / Q000737:chemistry* / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics*; D019204:GTP-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D064547:Myxovirus Resistance Proteins; D009690:Nucleic Acid Conformation; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics*
D003062:Codon; D064547:Myxovirus Resistance Proteins; D012333:RNA, Messenger; D019204:GTP-Binding Proteins