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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 韓玉山(Yu-San Han) | |
dc.contributor.author | Hui-Ling Hsu | en |
dc.contributor.author | 許蕙齡 | zh_TW |
dc.date.accessioned | 2021-06-17T01:16:04Z | - |
dc.date.available | 2023-10-25 | |
dc.date.copyright | 2020-08-24 | |
dc.date.issued | 2020 | |
dc.date.submitted | 2020-08-17 | |
dc.identifier.citation | 方苡帆,應用重組介白素-6於點帶石斑魚之抗病機轉研究,國立成功大學生物科技研究所碩士論文,2013。 陳信宏,點帶石斑魚介白素-6之基因轉殖、蛋白表現及功能分析,國立成功大學生物科技研究所碩士論文,2011。 蔡庚辰,利用螢光共振能量轉移技術(FRET)證明點帶石斑魚TNF-α1及TNF-α2之間的交互作用,國立成功大學生物科技研究所碩士論文,2016。 蘇建宏,在不同免疫刺激下點帶石斑其IL-6, IL-6R和gp130的訊息傳遞,國立成功大學生物科技研究所碩士論文,2015。 林伯峰,點帶石班魚兩腫瘤壞死因子-α (TNF-α1、TNF-α2) 蛋白功能性分析探討,國立成功大學生物科技研究所碩士論文,2012。 張晉榮,比較巴斯德桿菌與坎氏弧菌之共同免疫原對於石斑魚抵抗坎氏弧菌感染之交叉保護能力研究,國立成功大學生物科技研究所碩士論文,2011。 Abós, B., T. Wang, R. Castro, A. G. Granja, E. Leal, J. Havixbeck, A. Luque, D. R. Barreda, C. J. Secombes and C. Tafalla (2016). 'Distinct differentiation programs triggered by IL-6 and LPS in teleost IgM+ B cells in the absence of germinal centers.' Scientific reports 6(1): 1-16. Abe, K., M. Hirai, K. Mizuno, N. Higashi, T. Sekimoto, T. Miki, T. Hirano and K. Nakajima (2001). 'The YXXQ motif in gp 130 is crucial for STAT3 phosphorylation at Ser727 through an H7-sensitive kinase pathway.' Oncogene 20(27): 3464-3474. Abeywardena, M., W. Leifert, K. Warnes, J. Varghese and R. 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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66972 | - |
dc.description.abstract | gp130是非常重要的穿膜蛋白,可傳遞IL-6所誘發的免疫訊息傳遞。當IL6與IL-6r結合時,會進一步跟膜上的gp130結合形成複合物。該複合物可經由磷酸化進而活化下游的STAT-3訊號,隨後的信號傳遞誘發下游基因開始進行傳遞免疫訊息。在本篇研究中,點帶石斑於gp130全長序列首先經由circular DNA/RNA as template RACE方法進行選殖。其circular DNA/RNA as template RACE與市售的RACE試劑盒相比,circular DNA/RNA as template RACE可以降低RNA降解程度,並降低其時間及金錢成本。取得點帶石斑gp130開放閱讀框序列,全長約2694bp,可編碼899個氨基酸。經由MEGA5軟體,分析其gp130與其他魚類之gp130以及哺乳類之gp130氨基酸序列進行多序列分析,經親緣關係樹分析,將與gp130同一家族的其他蛋白-colony-stimulating factor receptor (CSFR)、Interleukin-31 Ra (IL-31Ra)、leukemia inhibitory factor receptor (LIFR) 以及oncostatin M receptor (OSM) 當作外群進行分析。並分析其穿膜區,膜內區和膜外區的位置。結果發現所選殖基因與gp130最為接近。為了進一步探討其gp130的生物功能為何,將其gp130膜外區與載體連接並轉殖入大腸桿菌中進行重組蛋白表現。接著進行點帶石斑的抗血清製備。在這項研究中,我們已製備許多重要的基礎研究,這些研究及抗血清製備將有助於未來使用免疫共沉澱的方法,以探討IL-6,IL-6R和gp130三者蛋白之間的相互作用。 | zh_TW |
dc.description.abstract | Glycoprotein130 is an essential transmembrane protein, it plays a critical role in IL-6 immune signal transduction. When IL6 combined with IL-6R, and the combined protein were further bind with gp130 on the target cell membrane, forming a complex. The complex activating the signaling transduction by phosphorylated signal transducer and activator of transcription 3; STAT-3, the subsequent signal transduction will turn on downstream genes which induce immune reactions. In this research, full length grouper gp130 sequence was first cloned through circular DNA/RNA as template RACE. Compare with commercial RACE kit, circular DNA/RNA as template RACE can avoid RNA degradation, more convenience and more time and money save. The full length 2694 bp grouper gp130 open read frame sequence was obtained, which estimated encode an 899 amino acid protein. Secondary, the Phylogenetic analysis was performed by multiple alignment of grouper gp130 with available fish gp130 and mammal gp130 amino acid sequence.by using MEGA5 software, leukemia inhibitory factor receptor (LIFR) and oncostatin M receptor (OSM) was using as the out group in the analysis. In the analysis all contain to segregate the transmembrane domain, intramembrane domain and outer membrane domain. In order to further verified the possible biofunction of gp130, gp130 outer membrane domain was further subcloning in to a E. coli expression plasmid and the recombinant protein was expression. Then the antiserum against grouper was also prepared. In this study, we prepared all the important tools, that will help to discover the interactions of IL-6, IL-6R and gp130 proteins in the future. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T01:16:04Z (GMT). No. of bitstreams: 1 U0001-1608202022162500.pdf: 3548193 bytes, checksum: 04261d51d5372b75129abc53af99ed9b (MD5) Previous issue date: 2020 | en |
dc.description.tableofcontents | 致謝 1 中文摘要 2 英文摘要 3 一、 研究背景 4 1.1點帶石斑 (Epinephelus coioides) 4 1.2 魚類免疫系統 4 1.3 介白素6 (Interleukin-6, IL-6) 6 1.4 介白素-6受體 (Interleukin-6 receptor, IL-6R) 7 1.5 醣蛋白130 (glycoprotein 130, gp130) 7 1.6 gp130的魚類研究 8 1.7 點帶石斑IL-6的相關研究 9 二、 研究目的 11 三、 材料與方法 12 3.1 材料方法的文獻回顧 12 3.1.1 未知基因全長選殖 12 3.1.2 互補DNA端點快速增幅反應 Rapid amplification of cDNA ends (RACE) 12 3.2 基因序列全長定序 15 3.3 點帶石斑魚gp130基因質體構築 19 3.4 重組蛋白的表現、鑑定及純化 22 3.5 鼠抗點帶石斑魚細胞外部分gp130抗體的製備 26 四、 實驗結果 29 4.1 點帶石斑gp130 基因全長定序 29 4.2 gp130以及其他同家族蛋白親緣關係樹分析 30 4.3 蛋白質結構模擬分析 30 4.4 gp130胞外區蛋白表現 31 4.5 小鼠rgp130-1、rgp130-2與rgp130-3專一性多株抗體製造 32 五、 討論 33 5.1 點帶石斑gp130 5’端基因定序 33 5.2 點帶石斑gp130和其他魚類相似度以及功能之比較 34 5.3 gp130構型研究 35 5.4 gp130重組蛋白的製造及免疫應用性 36 5.5 總結 37 六、 參考文獻 38 七、 圖表 48 | |
dc.language.iso | zh-TW | |
dc.title | "點帶石斑(Epinephelus coioides)之glycoprotein130 (gp130)基因選殖,細胞外片段重組蛋白表現及抗血清製作" | zh_TW |
dc.title | Red Spotted Grouper (Epinephelus coioides) Glycoprotein 130 Gene Cloning, Extra Cellular Domain Recombinant Protein Expression, and Antiserum Preparation | en |
dc.type | Thesis | |
dc.date.schoolyear | 108-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 林翰佑(Han-You Lin) | |
dc.contributor.oralexamcommittee | 龔紘毅(Hong-Yi Gong),邱品文(Pin-Wen Chiou) | |
dc.subject.keyword | 醣蛋白130,點帶石斑,抗血清,免疫系統, | zh_TW |
dc.subject.keyword | gp130,grouper,immune system,RACE,anti-serum, | en |
dc.relation.page | 80 | |
dc.identifier.doi | 10.6342/NTU202003623 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2020-08-18 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 漁業科學研究所 | zh_TW |
顯示於系所單位: | 漁業科學研究所 |
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U0001-1608202022162500.pdf 目前未授權公開取用 | 3.47 MB | Adobe PDF |
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