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目的探讨丙型肝炎病毒(HCV)核心蛋白对转录因子ETF和TxRE活性的调控作用。方法对于瞬时转染p IRES-Core/Neo载体的L02细胞采用双荧光素酶报告基因系统检测核心蛋白对转录因子ETF和TxRE活性影响,并提取2株细胞的核蛋白和RNA,分别以Western blot试验和实时荧光定量PCR检测2种转录因子的表达量。结果双荧光素酶报告基因检测在L02-Core和L02-Neo细胞中转录因子ETF和TxRE的活性比值分别为2.981和3.063。Western blot试验灰度值分析2株细胞的核蛋白提取物中这2种蛋白表达量的比值分别为1.510和2.717。实时荧光定量PCR检测这2株细胞中ETF和TxRE的核酸水平比值分别为4.123和3.703。结论在表达HCV核心蛋白的L02细胞中,转录因子ETF和TxRE转录表达水平及转录活性均较高,HCV核心蛋白可能具有增强转录因子ETF和TxRE转录活性的作用。
Objective To investigate the regulation of hepatitis C virus (HCV) core protein on the transcription factor ETF and TxRE activity. Methods L02 cells transiently transfected with pIRES-Core / Neo vector were used to detect the effect of core protein on ETF and TxRE activity of transcription factors and to extract the nuclear and RNA of the two cell lines. Western Blot Test and real-time fluorescence quantitative PCR detection of two transcription factor expression. Results The dual luciferase reporter assay showed that the ratios of ETF to TxRE in L02-Core and L02-Neo cells were 2.981 and 3.063, respectively. Western blot analysis of the gray value of the two cell nuclear extracts of the two protein expression ratio of 1.510 and 2.717 respectively. Real-time fluorescence quantitative PCR detection of these two strains ETF and TxRE nucleic acid levels were 4.123 and 3.703. CONCLUSION: The transcriptional level of ETF and TxRE in L02 cells expressing HCV core protein are high, and the transcriptional activity of ETRE is high. The HCV core protein may enhance the transcriptional activity of ETF and TxRE.