| Summary: | 碩士 === 國立陽明大學 === 微生物暨免疫學研究所 === 88 === The double-spliced RNA (dsp-RNA) is a naturally produced RNA by HBV (10). From previous studies, it has been demonstrated that the double-spliced RNA, pCMV/DS/3081, played an inhibitory role in the expression of HBV genes and this inhibition was regulated at the level of the transcription initiation (29). In this thesis, we would like to study further on the inhibitory role of the double-spliced RNA in HBV. The results are as follows: First, it is demonstrated that the 5’ end of transcript from pCMV/DS/3081, in which 162 nucleotides was transcribed from the putative transcription start site of CMV promoter, is not essential for the inhibitory effect. Also the inhibitory effect of the dsp-RNA could be observed by 24 hours after transfection. Second, five deletion clones were used to map the important regions for the inhibition. The data indicate that the 5’ region of the double-spliced RNA is crucial to the inhibitory activity. Finally, luciferase gene that was controlled either by single transcription element (TATA box) or complex up-regulatory cis-elements (authentic cytomealovirus immediate early promoter). We found that the dsp-RNA exhibited similar inhibitory activity in both systems. This results suggest that dsp-RNA inhibits the gene expression through its interaction with common trancriptional factors of TATA box.
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