Detection of EBER-2 in infact EBV-trasnsformed lymphocytes

• Main Authors: Gou Jerm-Feng, 高振峰
• Other Authors: Lin Shu-Bin
• Format: Others
• Language: zh-TW
• Published: 1994
• Online Access: http://ndltd.ncl.edu.tw/handle/30098498364193342237

碩士 === 國立臺灣大學 === 醫事技術學系 === 82 === Oligonucleotides can specifically hybridize the complementary sequences on DNA or RNA, therefore have been used to detect the specific fragments of genes, or regulate the expression of the genes, thus are recognized as potentially powerful tools in gene therapy.In this thesis, oligonucleoside methl phosphonate (OMP) and phosphorothioate oligonucleotides (PTO) were used to detect the specificc RNA in cells. Fluorescein labeled oligonucleoside methyl phosphonates (FOMP) and phosphorothioate oligonucleotides (FPTO) are synthesized to detect the specific RNA --- EBERs (EBV encoded RNA) in lymphocytes. We observed the distribution of fluorescence intensity by flow cytometer. We observed the accumulation of FOMP and FPTO complementary to EBER-2 in the nucleus of the cells containing EBERs, and they exist in the cytoplasm of the cells not containing EBERs. FOMP and FPTO not complementary to EBER-2 are only discovered in the cytoplasm in cells. FOMP and FPTO complementary to poly A tail exist in the whole cells. The fluorescence intensity of FOMP complementary to EBER-2 in the cells containing EBERs is higher than the cells not containing EBERs. The ratio of fluorescence intensity is 1.8. On the contrary, the fluorescence intensity of FOMP complementary to EBER-2 in the cells containing EBERs is much higher than the cells not containing EBERs. is much lesser than HR-1. It will enhance the fluorescence intensity by treating cells with monensin. The effect of monensin is different with the cell types and the kinds of oligonucleotide analogues. Both FOMP and FPTO can released from the cells, and reach equilibrium. The release rate of FOMP is much faster than FPTO, and the oligonucleotide analogues without target sequence in cells is released from cells much faster than the oligonucleotide analogus with target sequence in cells.