[Codon optimization of the H5N1 influenza virus HA gene gets high expression in mammalian cells].
In order to improve the expression of human avian influenza virus hemagglutinin (HA) and meet the pandemic influenza vaccine needs, we optimized and synthesized the whole length of HA gene of H5N1 (A/Anhui/1/2005) influenza virus in accordance with the human's codon preference, and inserted it to the eukaryotic expression vector pDC315 to construct a eukaryotic expression vector pDC315-Mod. HA. This plasmid and the eukaryotic expression vector pDC315-Wt. HA containing wild HA gene were transfected into 293T cells respectively to compare the expression of HA protein. The results showed that according to the comparison and identification by indirect immunofluorescence assay and Western blot test, the expression of HA protein in 293T cells was significantly improved after codon optimization. This laid a foundation for pandemic influenza vaccine research.
State Key Laboratory for Infectious Disease Prevention and Control, Chinese National Influenza Center, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China. kuibiao@gmail.com
Kui-Biao
KB
Li
Xiao-Guang
XG
Zhang
Jing
J
Ma
Xiao-Jun
XJ
Jia
Min
M
Wang
Jie
J
Dong
Xiao-Mei
XM
Zhang
Hong
H
Xu
Yue-Long
YL
Shu
D015153:Blotting, Western; D002460:Cell Line; D003062:Codon; D019084:Fluorescent Antibody Technique, Indirect; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics*; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics*; D010957:Plasmids
D003062:Codon; D019267:Hemagglutinin Glycoproteins, Influenza Virus