Construction of pseudoviruses containing a partial NS1 gene of Japanese encephalitis virus
Construction of pseudoviruses containing a partial non-structural protein 1(NS1)gene of Japanese encephalitis virus(JEV)with nuclease resistance for application as quality control reagents in laboratory assay.Firstly,the plasmid pET-32a-MS2 underwent optimization and modification.Secondary,fragments of the partial NS1 gene of the JEV,along with inter-nal reference genes from mosquitoes and humans(mosquito 18S RNA and human GAPDH),were inserted into this plasmid,resulting in the construction of the recombinant plasmid pET-MS2-JEV.Subsequent steps involved the expression and purifica-tion of pseudoviruses.The construction of the recombinant plasmid pET-MS2-JEV yielded pseudoviruses,which obtained resil-ience against nucleases and various physical and chemical factors.These pseudoviruses exhibited stability when stored at various temperature conditions,including 4 ℃ and-20 ℃.This study accomplished the production of pseudoviruses characterized by outstanding stability,thus serving as reliable quality control reagents for JEV laboratory assays.
Japanese encephalitis virusbacteriophage MS2pseudovirus