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为了研究干扰素刺激基因1(Interferon-induced protein with tetratricopeptide repeats 1,IFIT1)在蓝舌病病毒1型（Bluetongue virus serotype 1,BTV1）感染复制过程中的作用，首先利用实时定量PCR检测到BTV1感染绵羊睾丸细胞后IFIT1基因的转录水平明显升高，利用RT-PCR方法扩增获得羊IFIT1基因，测序后进行生物信息学分析，将其克隆到质粒载体pcDNA3.1/（+）上，构建重组表达质粒pcDNA3.1-OV-IFIT1，将其转染BHK-21细胞，观察到IFIT1基因在细胞内的成功表达，然后利用BTV1感染质粒pcDNA3.1-OV-IFIT1转染的细胞，从病毒的mRNA转录、蛋白表达和病毒滴度的变化评价IFIT1对BTV1复制的影响。结果显示，IFIT1在细胞中的过表达可显著抑制BTV1复制，相反，敲低IFIT1的表达可促进BTV1的复制。本研究首次报道了干扰素刺激基因IFIT1在BTV1感染复制过程中的作用，这将有助于揭示BTV1和宿主细胞IFIT1的相互作用机制，同时也为抗病毒药物研发提供了理论指导。
Abstract：To investigate the role of IFIT1 in the replication of bluetongue virus, real-time quantitative PCR was performed to detect the transcription level of IFIT1 in primary sheep testicular cells infected with bluetongue virus 1(BTV1). The ovine IFIT1 gene was amplified with RT-PCR and sequenced, and then analyzed with bioinformatics. The IFIT1 gene was subcloned to pcDNA3.1/（+） and the recombinant expression plasmid pcDNA3.1-OV-IFIT1 was constructed. The expression plasmid was transfected into BHK-21 cells. The expression of IFIT1 protein in the transfected cells was observed. The transfected cells were infected with BTV1 to evaluate the effect of IFIT1 on BTV1 replication, according to changes in viral mRNA transcription, protein expression, and viral titer. The results showed that IFIT1 overexpression resulted in a significant inhibition of BTV1 replication, whereas an IFIT1 knockdown in MDOK cells increased BTV1 replication. This study is the first to explore the role of IFIT1 during BTV1 infection, which helps to reveal the interaction between BTV1 and IFIT1, and also provides theoretical guidance for the development of antiviral drugs.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.004425
中图分类号:S852.65
引用信息:
[1]张国芮,康棣,王轩莹,等.干扰素刺激基因IFIT1抑制蓝舌病病毒的复制[J].病毒学报,2023,39(06):1660-1669.DOI:10.13242/j.cnki.bingduxuebao.004425.
基金信息:
国家重点研发计划项目（项目号：2021YFD1800500）,题目：动物疫病口岸检疫与风险防范技术研发及应用; 甘肃省基础研究创新群体项目（项目号：22JR5RA024）,题目：虫媒与虫媒病基础研究创新群体; 甘肃省科技厅项目（项目号：22CX8NA011）,题目：肉牛寄生虫病防控技术集成及示范应用; 国家产业技术体系项目（项目号：CARS-37）,题目：国家肉牛牦牛产业技术体系; 农业科技创新工程（项目号：CAAS-ASTIP-2016-LVRI）,题目：外寄生虫与虫媒疫病~~
2023-11-22
2023-11-22
2023-11-22