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Potent and specific inhibition of SARS-CoV antigen expression by RNA interference
Author(s): 
Pages: 166-169
Year: Issue:  3
Journal: CHINESE JOURNAL OF INFECTIOUS DISEASES

Keyword:  RNA干扰SARS病毒基因疗法基因表达;
Abstract: 目的构建绿色荧光蛋白(GFP)与严重急性呼吸综合征(SARS)冠状病毒 N蛋白的融合表达载体pEGFP-C1-N以及针对SARS冠状病毒 N基因的小发卡型shRNA表达载体pshRNA-N,观察shRNA对SARS冠状病毒 N基因复制和表达的影响.方法将构建成功的pEGFP-C1-N与psh RNA-N共转染293细胞,于转染后24、48、72 h观察GFP表达的强弱,采用Western Blot分别检测GFP与N蛋白的表达,逆转录PCR检测N基因的mRNA.结果 pshRNA-N作用组绿色荧光强度明显弱于未干扰组,Western Blot和逆转录PCR结果也证实pshRNA-N对N基因和N蛋白的表达有明显抑制作用,而无关序列的shRNA无此作用.结论针对SARS冠状病毒 N基因的shRNA具有显著和特异的抑制N蛋白表达的作用.
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