中国兽医科学2024，Vol.54Issue(5)：569-576,8.DOI:10.16656/j.issn.1673-4696.2024.0068
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
Construction of 293T cell line overexpressing JMJD6 gene and evaluation of its incremental efficiency against vesicular stomatitis virus
摘要
Abstract
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.关键词
JMJD6/水疱性口炎病毒/慢病毒包装/过表达细胞系/干扰素Key words
JMJD6/vesicular stomatitis virus/lentivirus packaging/overexpressed cell lines/in-terferon分类
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黄梦瑶,张伟,郑海学,杨帆,杨洋,邵文华,王家丽,吕岳芹,赵晓义,陈治彤,曹伟军..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[J].中国兽医科学,2024,54(5):569-576,8.基金项目
国家自然科学基金项目(32102639,32072831) （32102639,32072831）
国家重点研发计划项目(2021YFD1800300) （2021YFD1800300）
甘肃省杰出青年基金项目(21JR7RA026) （21JR7RA026）
兰州大学中央高校基本科研业务费专项资金(lzujbky-2022-ey20) （lzujbky-2022-ey20）
"十四五"广东省农业科技创新十大主攻方向"揭榜挂帅"项目(2023SDZG02) （2023SDZG02）
国家生猪产业技术体系(CARS-35). （CARS-35）
