李嘉哲2006-07-262018-07-112006-07-262018-07-112003http://ntur.lib.ntu.edu.tw//handle/246246/23617D 型肝炎病毒錄製二種同類型的D 型抗原。小D 型抗原為D 型肝炎病毒複 製所必須，而大D 型抗原比小D 型抗原多了C 端19 個胺基酸，不但可抑制病毒 複製且為病毒組合所必須。目前已知D 型抗原可與RNA 聚合脢II 、核仁磷化蛋 白B23 及nucleolin 結合，且經由此結合而調節病毒複製。但是D 型抗原參與 病毒複製的詳細機轉仍有待探討。大部分的蛋白質交互作用皆用酵母菌二雜交 法來研究。此法只能探知二種蛋白質的交互作用。最近標籤蛋白吸附純化法加 上質譜儀分析被成功應用於酵母菌的研究。有上千的多蛋白質複合體的個別蛋 白被詳細區分出來。我們試了幾種標籤載體來測試與D 型抗原形成複合體的細 胞蛋白。結果以pCMV-TAP 為最理想。目前已找到幾種蛋白，正做質譜儀分析中。Hepatitis delta virus (HDV)encodes two isoforms of delta antigens (HDAgs).The small form of HDAg (SHDAg)is required for HDV RNA replication,while the large form of HDAg (LHDAg),with additional C- terminal 19 amino acids,inhibits the viral replication and is required for viral assembly.Although HDAg was found to bind to RNA polymerase II,nucleolar phosphoprotein B23,and nucleolin and modulate HDV RNA replication,the detail mechanism and the role of HDAg in viral replication remain to be elucidated.Most protein interaction has been studied with yeast two-hybrid assay,which only detects interaction between two proteins.Recently,tagged-protein purification combined with mass spectrometry was applied successfully to characterize hundreds of distinct multiprotein complexes in the yeast.We had tried several tagged-vector and found pCMV-TAP,which contains proteinA and calmodulin was the best.we had found some proteins,which are characterized by mass spectrometry.application/pdf25026 bytesapplication/pdfzh-TW國立臺灣大學醫學院內科D 型抗原標籤蛋白吸附純化法質譜儀delta antigentagged-protein affinity purificationmass spectrometry[SDGs]SDG3reporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/23617/1/912315B002011.pdf