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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 吳岳隆(Yueh-Lung Wu) | |
| dc.contributor.author | Yu-Wei Chen | en |
| dc.contributor.author | 陳昱瑋 | zh_TW |
| dc.date.accessioned | 2021-06-15T12:25:47Z | - |
| dc.date.available | 2018-08-24 | |
| dc.date.copyright | 2016-08-24 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-08-11 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49890 | - |
| dc.description.abstract | 桿狀病毒可感染超過六百種無脊椎動物並具有高度的寄主專一性，因桿狀病毒具有高度寄主專一性，而被認為是安全的生物實驗材料，大量應用於生技產業做為外源基因蛋白質表現載體、疫苗生產與蟲害控制生物製劑。加州苜蓿夜蛾核多角體病毒 (AcMNPV) 為商品化且全球廣泛使用的桿狀病毒，在基因體層級上AcMNPV與家蠶核多角體病毒 (BmNPV) 具有極高度的同源性，但寄主範圍沒有重疊的現象。因此我們好奇兩種如此相似的病毒為何在同一寄主上能有如此大的感染差異。本研究將藉由次世代定序剖析病毒與寄主間的交互作用，分析家蠶細胞株 (BmN) 受AcMNPV 與 BmNPV感染後寄主轉錄體的調節模式，篩選可能參與交互作用並影響寄主專一性之因子。而在轉錄體的比較中可發現寄主在面對兩個相似病毒仍表現出不同的反應，且應用小片段核酸干擾 (RNAi) 驗證免疫基因確實抑制AcMNPV在家蠶細胞上的複製。另一方面，小片段核糖核酸 (Small RNA) 在許多研究中證實參與寄主專一性，因此我們掃描家蠶細胞內生性微小核酸 (MicroRNA) 在受病毒感染後表現的上升狀況，並結合次世代定序寄主基因表現，預測病毒誘導之MicroRNA與寄主細胞轉錄體間存在的交互作用。 | zh_TW |
| dc.description.abstract | Baculoviruses are insect-specific DNA viruses with restricted host range, which serve as viral vectors for bioindustry applications such as foreign gene expression, vaccine production, and pest control. As an example, Autographa californica nucleopolyhedrovirus (AcMNPV), a commercially available and widely used baculovirus prototype, can infect 39 species in 13 families. Another example would be Bombyx mori nucleopolyhedrovirus (BmNPV), a major pathogen of silkworms which has developed high host specificity to Bombyx mori. Interestingly, on a genomic level, the AcMNPV and BmNPV are highly homologous, but they share no overlapping host range. These two similar viruses have extremely different infection outcomes in Bombyx mori. We theorize that the determination of host specificity may depend on virus-host interactions, and that several genes may be involved in determining host specificity. Therefore, we used next-generation sequencing (NGS) to analyze transcriptome responses of hosts to these viruses. A transcriptome library was constructed, annotated, and grouped after sequence assembly. A comparison of gene expressions shows several significant differences in the gene expression profiles of BmNPV and AcMNPV, especially in cases where genes involved in immune responses were verified by RNA interference. In addition, studies have shown that small RNA plays a role in virus-host interaction, and further determines host specificity. Therefore, we screened microRNA induced by AcMNPV infection, and then combined NGS data from cellular gens to predict possible regulation networks. The manipulation of virus-host specificity could provide a breakthrough for the application of baculovirus in protein expression systems and in the development of bio-control agents. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T12:25:47Z (GMT). No. of bitstreams: 1 ntu-105-R03632008-1.pdf: 6090249 bytes, checksum: 8787390a304397b3729bb3c9aa082979 (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | Chapter 1 Introduction 1
Chapter 2 Materials and methods 7 2.1 Cell lines and viruses 7 2.2 Southern blot hybridization 7 2.3 Recombinant virus construction 8 2.4 RNAi experiments and transfection 8 2.5 Quantitation of gene expression 9 2.6 cDNA library construction and Illumina sequencing 10 2.7 Differentially expressed transcripts annotation and quantification 11 2.8 Gene ontology analysis 11 2.9 Cellular miRNA Microarray assay 11 2.10 miRNA target prediction 12 Chapter 3 Results 13 3.1 Cross infection of two closely related viruses demonstrates significant differences in host range 13 3.2 The viral gene expression was inhibited at a later time period in the nonpermissive cells 14 3.3 Assembly and annotation of B. mori cellular transcriptome reveal expression pattern divergence 15 3.4 Gene ontology analysis and host responses to virus infection 16 3.5Knockdown of immune gene facilitates vABhGFP replication in BmN cells 18 3.6 MicroRNA expression profile and interaction network against cellular genes 19 Chapter 4 Discussion21 References 29LIST OF FIGURE Figure 1. AcMNPV replication is strongly suppressed in BmN cells. 37 Figure 2. Late and very late gene expression was abolished by vABhGFP infected BmN cells. 38 Figure 3. Mapping rate and differentially expressed transcripts analysis of transcriptome. 39 Figure 4. Gene ontology analysis of differentially expressed gene among different molecule function (MF) at 48 hpi. 40 Figure 5. Knockdown of silkworm immune genes increase vABhGFP infection in BmN cells.42 Figure 6. Cellular miRNA expression profile analysis upon vABhGFP or BmNPV infected BmN cells. 44 Figure 7. Interaction network between cellular gene repressed by vABhGFP and miRNA induced by vABhGFP. 45 LIST OF TABLE Table 1. Modestly expressed genes are up-regulated by both vABhGFP and vBBhGFP. 46 Table 2. Top 20 genes are up-regulated by vBBhGFP.47 Table 3. Top 20 genes are down-regulated by vABhGFP.48 Table 4. Top 20 genes are up-regulated by vABhGFP.49 Table 5. Top 20 genes are down-regulated by vABhGFP.50 Table 6. Differentially expressed genes are up-regulated by vABhGFP as well as down-regulated by vBBhGFP. 51 | |
| dc.language.iso | en | |
| dc.subject | 寄主專一性 | zh_TW |
| dc.subject | 次世代定序 | zh_TW |
| dc.subject | 桿狀病毒 | zh_TW |
| dc.subject | host specificity | en |
| dc.subject | Baculoviruses | en |
| dc.subject | Next generation sequencing | en |
| dc.title | 昆蟲與桿狀病毒基因表現的互動與競爭分析 | zh_TW |
| dc.title | Analyses of Integrations and competitions between genes expressed from host and baculovirus | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張麗冠(Li-Kwan Chang),趙裕展(Yu-Chan Chao),乃育昕(Yu-Shin Nai),吳宗遠(Tzong-Yuan Wu) | |
| dc.subject.keyword | 桿狀病毒,次世代定序,寄主專一性, | zh_TW |
| dc.subject.keyword | Baculoviruses,Next generation sequencing,host specificity, | en |
| dc.relation.page | 51 | |
| dc.identifier.doi | 10.6342/NTU201602101 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2016-08-11 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 昆蟲學研究所 | zh_TW |
| 顯示於系所單位： | 昆蟲學系 | |
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