- 期刊
聚合酶連鎖反應法偵測EB病毒去氧核醣酸（DNA）的甲基化
Detection of EB Viral DNA Methylation by Polymerase Chain Reaction
摘要
為建立探討EB病毒（Epstein-Barr virus）基因體甲基化的研究模式，選擇兩種EB病毒細胞株，同時以南方墨點法及聚合酶連鎖反應法探討EB病毒C起動子（promoter）附近DNA的甲基化情形。兩種方法都顯示B95-8為非甲基化，Rael為甲基化。本研究提供聚合酶連鎖反應法來分析EB病毒DNA的甲基化情況，此種方法比起傳統的南方墨點法有以下的優點：所需的DNA量只及傳統方法的千分之一，操作步驟較簡便及可針對特定的CCmGG位置加以分析。值得注意的是想要成功的運用此種模式需要適量的DNA，高效率的限制酶作用，適當的增幅次數及謹慎的結果判讀。
關鍵字
甲基化，EB病毒，聚合酶連鎖反應並列摘要
In order to establish the model for detecting EB viral DNA methylation status, two cell lines were selected for comparing the EB viral DNA CCmGG methylation of C promoter (Cp) by using Southern blot and polymerase chain reaction (PCR). Both methods showed that the 5’sequence of Cp of B95-8 was unmethylated but Rael was methylated. It proved that polymerase chain reaction can be used as a tool for detection of EB viral DNA methylation. The advantages of this model are as follows: 1) Only 1/1000 amount of DNA used in Southern blot method is required. 2) The procedures are easy and take less time. 3) It can be used for detecting specific CCmGG methylation site. Appropriate DNA amount, efficient activity of restriction enzyme and suitable PCR cycles are successful assurance for this assay.
延伸閱讀
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