论文部分内容阅读
为研究机体干扰素诱导基因对日本乙型脑炎(JEV)感染的作用,通过IFN-α和JEV刺激猪睾丸(ST)细胞,实时荧光定量PCR检测Mx1、Mx2、OAS1、OAS2、PKR和干扰素诱导基因-15(ISG15)表达情况,发现除PKR外,其他基因相对mRNA量都有不同程度上升,其中ISG15上升最为显著。ISG15在ST细胞中过表达,检测病毒基因组和滴度,结果表明:ISG15可显著抑制JEV增殖;shRNA靶向干扰ISG15,JEV病毒量上升,说明ISG15在体外对JEV的增殖有抑制作用。将JEV的C、M、E、NS1、NS2A、NS2B、NS3、NS4A、NS4B基因克隆入pcDNA3.1真核表达载体,转染ST细胞后E和NS3基因引起ISG15相对mRNA量显著上升,说明JEV的E和NS3基因参与诱导ISG15的表达。
To investigate the effect of interferon-inducible genes on Japanese JEV infection, pig testes (ST) cells were stimulated with IFN-α and JEV, and Mx1, Mx2, OAS1, OAS2, PKR and interference were detected by real-time fluorescence quantitative PCR The expression of ISG15 was found to be significantly increased in ISG15, except for PKR. Among them, ISG15 increased most significantly. ISG15 was overexpressed in ST cells, and the virus genome and titer were detected. The results showed that ISG15 could significantly inhibit the proliferation of JEV. ShRNA targeted interference with ISG15 and JEV increased the amount of ISG15, indicating that ISG15 could inhibit the proliferation of JEV in vitro. The eukaryotic expression vectors of C, M, E, NS1, NS2A, NS2B, NS3, NS4A and NS4B of JEV were cloned into the pcDNA3.1 eukaryotic expression vector. After transfection of ST cells, E and NS3 genes caused a significant increase in the relative amount of mRNA of ISG15, indicating that JEV The E and NS3 genes are involved in the induction of ISG15 expression.