首页> 外文期刊>Turkish journal of biology >Expression profiles of interferon-related genes in cells infected with influenza A viruses or transiently transfected with plasmids encoding viral RNA polymerase
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Expression profiles of interferon-related genes in cells infected with influenza A viruses or transiently transfected with plasmids encoding viral RNA polymerase

机译:用甲型流感病毒感染的细胞中的干扰素相关基因的表达谱或用编码病毒RNA聚合酶的质粒瞬时转染

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Influenza A viruses frequently change their genetic characteristics, which leads to the emergence of new viruses. Consequently, elucidation of the relationship between influenza A virus and host cells has a great importance to cope with viral infections. In this study, it was aimed to determine expression profiles of interferon response genes in human embryonic kidney 293 (HEK293) cells infected with human (A/WSN-H1N1) and avian influenza A viruses (duck/Pennsylvania/10218/84/H5N2) or transfected with plasmids encoding viral RdRP subunits and, to obtain clues about the genes that may be important for the viral pathogenesis. The HEK293 cells cultured in a 12-well plate were infected with influenza A viruses or transfected with plasmids encoding viral polymerase. Total RNA extraction and cDNA preparation were carried out with commercial kits. Qiagen 96-well-RT2 Profiler PCR Array plates designated for interferons response genes were used for quantitation of the transcripts. The relative quantities of transcripts were normalized with STAT3 gen, and the results were evaluated. Quantitative RT-PCR results showed that there are substantial differences of the interferon response gene transcription in cells infected with viruses or transfected with plasmids. A higher number of interferon-related genes were found to be downregulated in the cells infected with DkPen compared to WSN. On the other hand, significant differences in the expression profiles of interferon response genes were observed in the cells expressing viral PA protein. In particular, avian influenza PA protein was found to cause more aggressive changes on the transcript levels. Human and avian influenza A viruses cause a substantial change in interferon response gene expression in HEK293 cells. However, a higher number of genes were downregulated in the cells infected with avian influenza DkPen compared to WSN. It has been also concluded that the viral PA protein is one of the important viral factors affecting the transcript level of host genes.
机译:流感病毒经常改变其遗传特征,导致新病毒的出现。因此,阐明了流感病毒和宿主细胞之间的关系具有重要意义来应对病毒感染。在本研究中,旨在确定感染人胚胎肾脏293(HEK293)细胞中干扰素反应基因的表达谱(A / WSN-H1N1)和禽流感的病毒(Duck / Pennsylvania / 10218/84 / H5N2)或用编码病毒RDRP亚基的质粒转染,并获得关于对病毒发病机制可能重要的基因的线索。在12孔板中培养的HEK293细胞用流感病毒感染或用编码病毒聚合酶的质粒转染。通过商业试剂盒进行总RNA提取和cDNA制备。 QIAGEN 96- RT2用于干扰素响应基因指定的勘探物PCR阵列板用于定量转录物。用STAT3 GEN标准化转录物的相对量,并评估结果。定量RT-PCR结果表明,感染病毒或用质粒转染细胞中的干扰素响应基因转录存在显着差异。与WSN相比,发现在用DKPen感染的细胞中发现了较多的干扰素相关基因。另一方面,在表达病毒PA蛋白的细胞中观察到干扰素反应基因的表达谱的显着差异。特别是,发现禽流感PA蛋白导致转录水平的更具侵略性变化。人和禽流感,病毒导致HEK293细胞中干扰素反应基因表达的大量变化。然而,与WSN相比,在感染禽流感DKPen的细胞中下调了更高数量的基因。还有人们还得出结论,病毒PA蛋白是影响宿主基因的转录物水平的重要病毒因素之一。

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