Abstract
The purpose of this thesis is to develop a DNA-based chip for studying gene expression in Epstein-Barr virus (EBV), including probe design, target preparation, membrane hybridization and data analysis. We intent to use the DNA chip to measure the expression level of many EBV genes simultaneously in a single hybridization assay. In the preliminary process established by us, the cDNA samples hybridized to the nylon array are l led by incorporating fluorescein tagged nucleotides during reverse primers-primed reverse transcription of total RNA. We have utilized an enzyme-linked colorimetry detection and chemiluminescent (CDP-Star) detection system to quantify gene expression level. To prove the feasibility of the process, we have investigated the expression pattern of EBV gene in P3HR-1 cells induced by two lytic cycle inducers, transforming growth factor-β (TGF-β) and 12-o-tetradecanoylphorbol- 13-acetate (TPA). We have been able to detect the changes in the expression of some of the genes, e.g. BZLF1, an immediate-early gene. Our study, based on the assay set up to supply information about the regulation of EBV genes, humbly but firmly suggests that the EBV gene chip certainly can provide a rapid and effective analytical tool for monitoring EBV gene expression.中文摘要----------------------------------------2.英文摘要----------------------------------------3.壹、 前言-----------------------------------4.貳、 材料與方法----------------------------15.參、 結果----------------------------------26.肆、 討論----------------------------------31.伍、 參考文獻------------------------------38.陸、 圖表----------------------------------43.