EGS对HCMV UL54基因表达的抑制及对病毒复制的影响

发布时间：2018-03-17 11:25

本文选题：EGS　切入点：抗病毒　出处：《暨南大学》2007年硕士论文　论文类型：学位论文

【摘要】： 目的：本实验拟在前期研究的基础上，筛选和检测已经经过体外实验证明有活性的HCMV UL54基因特异的EGS(dEGS-T6和dEGS-T7)的胞内抗病毒活性。方法：本课题分为三部分：首先用流式细胞仪检测了FITC标记的EGS转染细胞的效率；第二，，在感染病毒的培养细胞中，用相对定量PCR来检测转染的EGS对HCMV的U54基因表达的影响；第三，转染EGS的细胞感染病毒后，通过检测培养上清液中病毒量的变化，制作病毒生长曲线，以此评价EGS对HCMV复制的抑制作用。结果：虽然在高浓度(100nM和80nM)时，EGG表现出对病毒基因表达的非特异性抑制，但是在转染60nM的EGS时，dEGS-T7能明显抑制细胞内病毒U54基因mRNA的表达量而dEGS-T6和无关dEGS-TK对U54基因mRNA的表达无明显影响。因此，选择60hM为最佳转染浓度进一步检测dEGS-T7对胞内HCMV复制的影响时发现，dEGS-T7可以使病毒感染细胞培养液中的病毒减少约67.27±0.10％～76.47±1.23％，而转染无关EGS后与未转染EGS的对照组培养液中的病毒数无明显减少。结论：通过本研究的筛选步骤，获得一种具有胞内抗HCMV活性的EGG序列dEGS-T7，从而为新型抗HCMV反义药物的进一步研究与开发奠定了基础。
[Abstract]:Aim: to screen and detect the intracellular antiviral activities of HCMV UL54 gene specific EGS(dEGS-T6 and dEGS-T7, which have been proved to be active in vitro. Methods: this study was divided into three parts: firstly, the efficiency of FITC labeled EGS transfected cells was detected by flow cytometry; secondly, the effect of transfected EGS on the expression of U54 gene of HCMV was detected by relative quantitative PCR in infected virus culture cells. Thirdly, after the cells transfected with EGS were infected with virus, the virus growth curve was made by detecting the changes of virus content in the culture supernatant, so as to evaluate the inhibitory effect of EGS on the replication of HCMV. Results: although at high concentrations of 100nM and 80nM), EGG showed a nonspecific inhibition on viral gene expression. However, dEGS-T7 could significantly inhibit the expression of U54 gene mRNA in the cells transfected with 60nM EGS, but dEGS-T6 and unrelated dEGS-TK had no significant effect on the expression of U54 gene mRNA. The optimal transfection concentration of 60hM was chosen to further detect the effect of dEGS-T7 on the replication of intracellular HCMV. It was found that dEGS-T7 could reduce the virus content in the culture medium of virus-infected cells about 67.27 卤0.10 and 76.47 卤1.23. but after transfection of EGS and control medium without EGS transfection, the effect of DEGS-T7 on HCMV replication was observed. There was no significant decrease in the number of viruses. Conclusion: a EGG sequence dEGS-T7 with intracellular HCMV activity was obtained through the screening step of this study, which laid a foundation for the further research and development of novel antisense drugs against HCMV.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R373

【相似文献】