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目的了解干扰素刺激基因15(ISG15)对严重发热伴血小板减少综合征病毒(SFTSV)复制的影响。方法1)ISG15基因克隆:通过逆转录聚合酶链式反应(RT-PCR)的方法,从Hela细胞中扩增获得ISG15基因,然后连接质粒形成ISG15表达载体。2)研究ISG15表达对SFTSV复制的影响:将ISG15质粒转染Hela细胞并过表达后,接种SFTSV,随后用荧光定量RT-PCR的方法观察ISG15对SFTSV复制的影响。结果成功构建ISG15表达质粒,并在Hela细胞内过表达;SFTSV成功在Hela细胞内增殖,同时过表达ISG15可以抑制SFTSV的复制。结论初步证明在Hela细胞中,过表达ISG15可以抑制SFTSV的复制,但具体机制还有待进一步探讨。
Objective To investigate the effect of interferon stimulating gene 15 (ISG15) on the replication of severe fever with thrombocytopenic syndrome virus (SFTSV). Methods 1) ISG15 gene cloning: The ISG15 gene was amplified from Hela cells by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated to form the ISG15 expression vector. 2) To investigate the effect of ISG15 expression on SFTSV replication: The ISG15 plasmid was transfected into Hela cells and overexpressed SFTSV, then the effect of ISG15 on SFTSV replication was observed by real-time fluorescence quantitative RT-PCR. Results The ISG15 expression plasmid was successfully constructed and was overexpressed in Hela cells. SFTSV successfully proliferated in Hela cells. Over-expression of ISG15 could inhibit the replication of SFTSV. Conclusion Preliminary evidence shows that in Hela cells, overexpression of ISG15 can inhibit the replication of SFTSV, but the specific mechanism remains to be further explored.