Development and Use of a Kinetical and Real-Time Monitoring System to Analyze the Replication of Hepatitis C Virus

• Material Type: 博士論文
• Title: Development and Use of a Kinetical and Real-Time Monitoring System to Analyze the Replication of Hepatitis C Virus
• Author/Editor: Li, Xiaoyu
• Author Heading: Li, Xiaoyu
• Publication, Distribution, etc.: MDPI (Multidisciplinary Digital Publishing Institute)
• Publication Date: 2022-08-05
• Publication Date (W3CDTF): 2022-08-05
• Alternative Title: C型肝炎ウイルスの複製を解析するための動態学的かつリアルタイムモニタリングシステムの開発と利用
• Degree grantor/type: 浜松医科大学
• Date Granted: 2022-09-16
• Date Granted (W3CDTF): 2022-09-16
• Dissertation Number: 甲第913号
• Degree Type: 博士（医学）
• Text Language Code: eng
• Subject Heading: hepatitis C virus; genome replication; bioluminescence profile; real-time monitoring; kinetical analysis
• Target Audience: 一般
• Note (General): doctoral; 医学系研究科; In microbiological research, it is important to understand the time course of each step in a pathogen’s lifecycle and changes in the host cell environment induced by infection. This study is the first to develop a real-time monitoring system that kinetically detects luminescence reporter activity over time without sampling cells or culture supernatants for analyzing the virus replication. Subgenomic replicon experiments with hepatitis C virus (HCV) showed that transient translation and genome replication can be detected separately, with the first peak of translation observed at 3?4h and replication beginning around 20h after viral RNA introduction into cells. From the bioluminescence data set measured every 30min (48 measurements per day), the initial rates of translation and replication were calculated, and their capacity levels were expressed as the sums of the measured signals in each process, which correspond to the areas on the kinetics graphs. The comparison of various HuH-7-derived cell lines showed that the bioluminescence profile differs among cell lines, suggesting that both translation and replication capacities potentially influence differences in HCV susceptibility. The effects of RNA mutations within the 50 UTR of the replicon on viral translation and replication were further analyzed in the system developed, confirming that mutations to the miR-122 binding sites primarily reduce replication activity rather than translation. The newly developed real-time monitoring system should be applied to the studies of various viruses and contribute to the analysis of transitions and progression of each process of their life cycle.
• Persistent ID (NDL): info:ndljp/pid/12694247; https://dl.ndl.go.jp/pid/12694247
• Collection: 障害者向け資料
• Collection (Materials For Handicapped People:1): テキストデータ
• Collection (particular): 国立国会図書館デジタルコレクション > デジタル化資料 > 博士論文; https://dl.ndl.go.jp/collections/A00014
• Acquisition Basis: 博士論文（自動収集）
• Date Accepted (W3CDTF): 2023-03-07T16:07:55+09:00
• Format (IMT): application/pdf
• Access Restrictions: 国立国会図書館内限定公開
• Service for the Digitized Contents Transmission Service: 図書館・個人送信対象外
• Availability of remote photoduplication service: 可
• Periodical Title (URI): http://hdl.handle.net/10271/00004267; https://hama-med.repo.nii.ac.jp/records/4145
• Data Provider (Database): 国立国会図書館 : 国立国会図書館デジタルコレクション; https://dl.ndl.go.jp

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