| Summary: | 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.
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