中国兽医科学2025，Vol.55Issue(11)：1421-1428,8.DOI:10.16656/j.issn.1673-4696.2025.0192
稳定表达KCTD1基因的PK15细胞系构建及其对FMDV和VSV复制影响的初步研究
Construction of PK15 cell line with stable expression of KCTD1 and its impacts on the replication of FMDV and VSV
摘要
Abstract
A cell line stably expressing potassium channel tetramerization domain-containing 1(KCTD1)was established using a lentiviral packaging system.This model was then used to preliminarily investigate the role of KCTD1 in the replication of foot-and-mouth disease virus(FMDV)and vesicular stomatitis virus(VSV),which contributes to a better understanding of the regulatory function of KCTD1 in viral replication.In this study,the recombinant plasmid PLV-puro-3 ×Flag-KCTD1-HA was constructed and transfected into HEK293T cells to analyze its effect on VSV replication.Subsequently,a PK15 cell line with stable KCTD1 overexpression(PK15/KCTD1)was successfully generated using lentivirus packaging technology.The replication levels of FMDV and VSV in the PK15/KCTD1 cell line were assessed by Western-blot,qPCR,and virus titer assays.The results demonstrated that in the PK15/KCTD1 cell line,the levels of FMDV protein,RNA load,and virus titer were significantly higher compared with the control group transfected with an empty vector.Similarly,the replication level of VSV was also significantly elevated in the PK15/KCTD1 cell line.In conclusion,this study successfully established a PK15/KCTD1 cell line and demonstrated that KCTD1 promotes the replication of FMDV and VSV.关键词
KCTD1/慢病毒包装/E3泛素连接酶/细胞系/口蹄疫病毒/水疱性口炎病毒Key words
KCTD1/lentivirus packaging/E3 ubiquitin ligase/overexpression cell line/foot-and-mouth disease virus/vesicular stomatitis virus分类
农业科技引用本文复制引用
张艳,董苏洁,张志东,李彦敏..稳定表达KCTD1基因的PK15细胞系构建及其对FMDV和VSV复制影响的初步研究[J].中国兽医科学,2025,55(11):1421-1428,8.基金项目
国家重点研发计划项目(2024YFD1800304) （2024YFD1800304）
