过表达JMJD6基因的293T细胞系的构建及对水疱性口炎病毒增殖效率的评价OA北大核心CSTPCD

In order to enhance the efficiency of vesicular stomatitis virus(VSV)proliferation within cells,this study employed a lentivirus packaging system to establish a HEK-293T/JMJD6 cell line that exhibited an overexpression of Jumonji domain containing protein 6(JMJD6).Subsequently,the replication disparities of VSV between HEK-293T/JMJD6 and HEK-293T cells were analyzed.To begin,a re-combinant plasmid pLV-puro-3×Flag-JMJD6 gene was constructed and co-transfected with auxiliary plasmids pMD2.G and psPAX2 into HEK-293T cells for the purpose of lentivirus packaging.Following this,HEK-293T cells were infected with the packaged lentivirus,and positive cells overexpressing the JMJD6 gene were obtained by puromycin selection.To assess the replication ability of VSV in the HEK-293T/JMJD6 cell line,Western-blot,indirect immunofluorescence,flow cytometry,and plaque assay were employed.The results demonstrated that the proliferation ability of vesicular stomatitis virus(VSV)in the HEK-293T/JMJD6 cell line was significantly greater compared to wild-type cells.Reverse transcription polymerase chain reaction(RT-PCR)analysis of interferon downstream gene expression in VSV-infected HEK-293T/JMJD6 cell lines and wild-type cells revealed that the HEK-293T/JMJD6 cell line suppressed the production of type Ⅰ interferon downstream genes induced by VSV.The establishment of the HEK-293T/JMJD6 cell line in this study effectively enhances VSV proliferation and inhibits the type Ⅰ interferon signaling pathway,thereby providing a basis for the identification of potential VSV vaccine candidate cell lines.