Construction and primary expression of eukaryotic expressing vectors of codon-optimized H1N1 influenza virus HA and HA1 gene.

【Objective】The study was done to construct influenza A virus H1N1-Mod.HAand H1N1 Mod.HA1eukaryotic expressing plasmids and test their expression in BHK-21 cells.【Method】HAand HA1 DNA sequences of H1N1 influenza A virus were optimized in accordance with human codon preference so as to enhance its protein expression in eukaryotic expressing system.Synthetic HAand HA1 genes were inserted into the eukaryotic expressing vector pSNAV,which constructed Eukaryotic Expressing Plasmids of Mod.HA and Mod.HA1. Mod.HAand Mod.HA1 eukaryotic expressing plasmids were transfected into BHK-21 cells.G 418 selection acquired monoclone.Immunofluorescence assay and Western blot detected expression in BHK-21 cells.【Result】It was confirmed that the construction of Mod.HAand Mod.HA1 eukaryotic expressing plasmids was made successfully.The 10 generations BHK-21 cells were tested with immunofluorescence and Western blot,which showed HAand HA1 were both stably and accu-rately expressed in BHK-21 cells.【Conclusion】The experiment successfully constructed eukaryotic expressing plasmids for Mod.HAand Mod.HA1,which could provide a foundation for the further study on the development of genetic engineering vaccine and the tested reagent of H1N1 influenza.