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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王重雄(Chung-Hsiung Wang) | |
| dc.contributor.author | Tai-Chuan Wang | en |
| dc.contributor.author | 王泰權 | zh_TW |
| dc.date.accessioned | 2021-06-13T08:19:55Z | - |
| dc.date.available | 2007-07-26 | |
| dc.date.copyright | 2005-07-26 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-07-19 | |
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Wang. 2005. Characterization and polyhedrin gene cloning of Lymantria xylina multiple nucleopolyhedrovirus. J. Invertebr. Pathol. 88: 238-246. Zanotto, P. M. D. A., B. D. Kessing, and J. E. Maruniak. 1993. Phylogenetic interrelationships among baculoviruses: evolutionary rates and host associations. J. Invertebr. Pathol. 62: 147-164. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/36865 | - |
| dc.description.abstract | 核多角體病毒感染細胞株之末期，會產生許多由多角體蛋白所包埋的封埋體病毒，根據細胞內形成多角體的多寡可分為繁多角體 (multiple polyhedra, MP) 和寡多角體 (few polyhedra, FP) 兩種。寡多角體的形成與是否能夠轉譯出 25 kDa 的 fp25k 基因有關。當 fp25k 基因發生缺失或是插入外來的 DNA 片段時，會使病毒在感染細胞後呈現 FP 的現象。榕樹透翅毒蛾核多角體病毒 (Perina nuda multiple nucleopolyhedrovirus, PenuMNPV) 與黑角舞蛾核多角體病毒 (Lymantria xylina multiple nucleopolyhedrovirus, LyxyMNPV) 的 fp25k 基因分別含有 624 與 651 個鹼基對，且榕樹透翅毒蛾核多角體病毒與黃杉毒蛾核多角體病毒 (Orgyia pseudotsugata multiple nucleopolyhedrovirus, OpMNPV) 較為相似；黑角舞蛾核多角體病毒則與吉普賽舞蛾核多角體病毒 (Lymantria dispar multiple nucleopolyhedrovirus, LdMNPV) 較為相似。榕樹透翅毒蛾與黑角舞蛾核多角體病毒的 fp25k 基因皆具有一個晚期和非常晚期基因的轉錄起始點 ATAAG，為桿狀病毒感染過程中晚期表現基因。FP25K 蛋白質經電腦預測發現具有 coiled-coil 和 putative actin-binding domains。根據 fp25k 基因所建構之親緣關係樹發現榕樹透翅毒蛾核多角體病毒屬於 group Ⅰ，而黑角舞蛾核多角體病毒屬於 group Ⅱ 的病毒群。在西方墨點法偵測榕樹透翅毒蛾核多角體病毒的 FP25K 蛋白發現，其為一個結構蛋白並且表現在感染的晚期。以間接免疫螢光法發現榕樹透翅毒蛾核多角體病毒的 FP25K 表現在細胞質與細胞核內。野生型的榕樹透翅毒蛾與黑角舞蛾核多角體病毒皆為 MP 的病毒株。但 LyG (LdMNPV-like virus) 病毒株感染吉普賽舞蛾細胞株後皆呈現 FP 的現象，此病毒的 fp25k 基因序列發生缺失現象，經由電腦分析轉譯結果顯示造成框架移動突變 (frameshift mutation)，在 Ly5 (LyxyMNPV) 病毒株為 MP 與 FP 共存的現象，分析發現具有野生型與缺失的 fp25k 基因，因此 fp25k 基因的突變將會直接影響到桿狀病毒的 FP 現象。 | zh_TW |
| dc.description.abstract | The nucleus of an infected cell by nucleopolyhedrovirus is filled with occlusion bodies (OBs) embeded by polyhedrin in the late infection phase. According to the numbers of polyhedra per cell, two types of cytopathic effect (CPE) can be divided, multiple polyhedra (MP; >10 OBs/cell) and few polyhedra (FP; < 10 OBs/cell). FP phenotype is associated with fp25k gene which encodes 25 kDa. When the fp25k sequence is deleted or inserted by the foreign DNA, it causes reducing numbers of OBs in the infected cells. The fp25k of Perina nuda multiple nucleopolyhedrovirus (PenuMNPV) and Lymantria xylina multiple nucleopolyhedrovirus (LyxyMNPV) contained 624 and 651 base pairs, respectively. The former is closely related to Orgyia pseudotsugata multiple nucleopolyhedrovirus (OpMNPV) and the later is to Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV). Both PenuMNPV and LyxyMNPV fp25k genes are late genes containing a transcriptional initiation site, TAAG, of late and very late gene motifs. FP25K protein possessed with coiled-coil and putative actin-binding domains by computer prediction. A phylogenetic tree was constructed based on fp25k sequences, PenuMNPV and LyxyMNPV belonged to group Ⅰ and group Ⅱ, respectively. FP25K protein was confirmed that it is a structure protein and expressed in late phase by Western blotting hybridization. Both PenuMNPV (Ө52) and LyxyMNPV (Ly2) in our lab strains displayed MP phenotype. LyG (LdMNPV-like virus) from infected Lymantria xylina exhibited FP phenotype in infected cells, and its fp25k was deleted 44 nucleotides comparing with the fp25k of LdMNPV. Ly5 strain exhibited two phenotypes, MP and FP in infected LD cells, its fp25k sequence contained wild and deleted type. Therefore, the expression of fp25k or deleted fp25k directly affected the CPE (MP or FP) phenotype of infected cells. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T08:19:55Z (GMT). No. of bitstreams: 1 ntu-94-R92632006-1.pdf: 686724 bytes, checksum: e6a3977d11bd76fd6aa606c16fafb1e8 (MD5) Previous issue date: 2005 | en |
| dc.description.tableofcontents | 中文摘要
英文摘要 目錄.....................................................i 表次.....................................................iv 圖次.....................................................v 壹、緒言.................................................1 貳、往昔研究.............................................3 參、材料與方法...........................................6 一、供試病毒與寄主.......................................6 二、病毒的感染...........................................6 三、增幅病毒 fp25k 基因部分片段及全長....................6 四、病毒 RNA 之萃取......................................7 五、三端與五端快速增幅互補 DNA...........................7 六、封埋體病毒與病毒粒子的純化...........................8 七、病毒基因組 DNA 之純化................................9 八、含有 fp25k 基因之病毒基因組片段之切割................9 九、探針 (probe) 的製備..................................10 十、南方墨點法 (Southern blot)...........................10 十一、選殖 (cloning).....................................11 十二、質體 DNA 小量純化..................................11 十三、細菌轉型作用 (transformation)......................11 十四、表現 PenuMNPV 與 LyxyMNPV 之 FP25K 重組蛋白........12 十五、聚丙烯醯胺膠體電泳 (SDS-PAGE)......................12 十六、PenuMNPV 之 FP25K 的時序性表現.....................12 十七、西方墨點法 (Western blot)..........................13 十八、病毒斑測試 (plaque assay)..........................13 十九、間接免疫螢光.......................................13 二十、FP25K 的蛋白質結構預測.............................14 二十一、親緣關係分析 (phylogenetic analysis).............14 肆、結果.................................................15 一、PenuMNPV、LyxyMNPV 與 LdMNPV-like 病毒感染細胞後之外表型.......................................................15 二、病毒 fp25k 部分基因選殖..............................15 三、PenuMNPV 與 LyxyMNPV 基因組經限制酶切割後含有 fp25k 之片段.....................................................15 四、PenuMNPV 之 fp25k 基因的三端與五端...................15 五、LyxyMNPV 之 fp25k 基因的三端與五端...................16 六、FP 25K 之蛋白質結構分析..............................16 七、表現 PenuMNPV 與 LyxyMNPV 的 FP25K 重組蛋白..........16 八、PenuMNPV 的 FP25K 之時序性表現.......................16 九、PenuMNPV 的FP25K 為一個結構蛋白......................17 十、間接免疫螢光抗體染色.................................17 十一、PenuMNPV、LyxyMNPV 與 LdMNPV-like 病毒之 fp25k 基因檢測.......................................................17 十二、親緣關係之分析.....................................17 十三、PenuMNPV 與 LyxyMNPV 與其他桿狀病毒 fp25k 基因和蛋白質相似度比較.............................................18 伍、討論.................................................19 陸、結論.................................................23 柒、參考文獻.............................................24 捌、圖表.................................................31 玖、致謝.................................................46 拾、附錄.................................................47 附錄一、建構演化樹所使用各種核多角體病毒之 GenBanK 的 accession number.........................................47 附錄二、本實驗中所使用的引子名稱與序列...................48 表次 表一、核多角體病毒的 fp25k 基因序列相似度的比較..........31 表二、核多角體病毒的 FP25K 蛋白質 identity 和 similarity 的比較.....................................................32 圖次 圖一、榕樹透翅毒蛾核多角體病毒、黑角舞蛾核多角體病毒與 LdMNPV-like 病毒感染細胞後所呈現的細胞病變現象..........33 圖二、增幅 fp25k 部分基因片段...........................34 圖三、以南方墨點法偵測榕樹透翅毒蛾核多角體病毒基因組中含有 fp25k 之限制酶片段......................................35 圖四、以南方墨點法偵測黑角舞蛾核多角體病毒基因組中含有 fp25k 之限制酶片段......................................36 圖五、榕樹透翅毒蛾核多角體病毒之 fp25k 序列.............37 圖六、黑角舞蛾核多角體病毒之 fp25k 序列.................38 圖七、電腦分析比對核多角體病毒的 FP25K 蛋白質結構.......39 圖八、利用細菌表現載體表現榕樹透翅毒蛾與黑角舞蛾核多角體病毒的 FP25K 重組蛋白.....................................40 圖九、榕樹透翅毒蛾核多角體病毒 FP25K 蛋白之時序性表現...41 圖十、西方墨點法偵測榕樹透翅毒蛾核多角體病毒之病毒粒子的結構蛋白..................................................42 圖十一、間接免疫螢光偵測榕樹透翅毒蛾核多角體病毒 FP25K 在細胞中的表現位置..........................................43 圖十二、研究中所使用榕樹透翅毒蛾核多角體病毒 (PenuMNPV, Ө52)、黑角舞蛾核多角體病毒株 (LyxyMNPV: Ly2, Ly5) 及 LdMNPV-like 病毒 (LyG) 之外表型與 fp25k 基因結構........44 圖十三、根據 fp25k 基因所建構之親緣關係樹...............45 | |
| dc.language.iso | zh-TW | |
| dc.title | 比較榕樹透翅毒蛾核多角體病毒與黑角舞蛾核多角病毒之 fp25k 基因和其在昆蟲細胞內的表現 | zh_TW |
| dc.title | Comparison of two fp25k genes from Perina nuda and Lymantria xylina multiple nucleopolyhedroviruses and their expression in insect cells | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 侯豐男(Feng-Nan Hou),路光暉(Kuang-Hui Lu),李松泰(Song-Tay Lee),吳文哲(Wen-Jer Wu) | |
| dc.subject.keyword | 樹透翅毒蛾核多角體病毒,黑角舞蛾核多角病毒,fp25k 基因, | zh_TW |
| dc.subject.keyword | Perina nuda multiple nucleopolyhedroviruses,Lymantria xylina multiple nucleopolyhedroviruses,fp25k gene, | en |
| dc.relation.page | 48 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2005-07-19 | |
| 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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