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通过RT-PCR的方法克隆H5N1亚型禽流感病毒NS1基因,并构建了真核表达载体pCMV-Myc/NS1。将此真核表达质粒转染肺腺癌细胞A549,48 h后,经Western印迹检测,NS1基因能在细胞中正确表达。经荧光显微镜、透射电镜观察和流式细胞仪检测,发现该株流感病毒的NS1蛋白可诱导肺腺癌细胞A549凋亡。
The NS1 gene of H5N1 avian influenza virus was cloned by RT-PCR and the eukaryotic expression vector pCMV-Myc / NS1 was constructed. After transfection of this eukaryotic expression plasmid into human lung adenocarcinoma A549 cells for 48 h, the NS1 gene was correctly expressed in the cells by Western blot. Fluorescence microscopy, transmission electron microscopy and flow cytometry showed that the NS1 protein of the influenza virus could induce the apoptosis of lung adenocarcinoma A549 cells.