Fig2G_RACE_YKT6_R2_LG364.tif

Xrn1 knock out A549 cells were infected with WT A/Puerto Rico/8/1984 (H1N1) ("PR8"), A/Tennessee/1-560/2009 (H1N1) (“H1N1pdm09” for 2009 pandemic H1N1) , or  A/Perth/16/2009 (H3N2) ("Perth"), or PA(∆X) mutants of these strains, or mock infected for 16 hours before RNA extraction. 5’RACE was then performed using primers specific for STOML2 or YKT6, ~250-300 nucleotides (nt) downstream of the predicted cut sites. The PCR products were run on an agarose gel. The predicted size of DNA bands coming from cut sites identified by PyDegradome are indicated by the red dotted arrows.

将Xrn1基因敲除（Xrn1 knockout）的A549细胞分别感染野生型（Wild Type, WT）A/波多黎各/8/1984（H1N1）（简称PR8）、A/田纳西/1-560/2009（H1N1）（即2009年大流行性H1N1毒株，缩写为H1N1pdm09）、A/珀斯/16/2009（H3N2）（简称Perth），以及上述毒株的PA(∆X)突变株，同时设置模拟感染对照组，感染16小时后提取RNA。随后，在预测剪切位点下游约250~300个核苷酸（nt）处，针对STOML2或YKT6基因使用特异性引物开展5'末端快速扩增（5'-Rapid Amplification of cDNA Ends, 5'RACE）实验；将所得聚合酶链式反应产物进行琼脂糖凝胶电泳，由PyDegradome鉴定的剪切位点对应的预期DNA条带大小以红色虚线箭头标注。