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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林峰輝(Feng - Huei Lin) | |
| dc.contributor.author | Chu-Li Peng | en |
| dc.contributor.author | 彭茱莉 | zh_TW |
| dc.date.accessioned | 2021-06-08T07:20:12Z | - |
| dc.date.copyright | 2008-07-26 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-07-25 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26672 | - |
| dc.description.abstract | 本實驗將利用明膠當作為一個新的基因載體，並做出粒徑為300nm左右的明膠奈米微粒。且於DNA的包覆率測試上，發現其包覆率可達到80％以上；佐以雷射奈米粒徑暨界面電位測量儀、穿透式電子顯微鏡證實此明膠奈米微粒的粒徑與穩定性。且經由DNase Ι的作用反應可以說明明膠奈米微粒具有保護DNA的能力。於HeLa細胞的轉染實驗中，發現明膠奈米微粒和市售商品LipofectaineTM 2000有幾乎相似的轉染效率，但是相對於市售商品LipofectaineTM 2000，明膠奈米微粒有較高的細胞活性和較低的細胞毒性。於轉染雞胚的實驗中，經由明膠奈米微粒轉染的雞胚其存活率約為80-100％，觀察被轉染四天後的雞胚也可以發現綠色螢光蛋白的產生。這說明了明膠奈米微粒的生物相容性的優勢在轉殖動物上是極具潛力的。 | zh_TW |
| dc.description.abstract | In thus study, we tried to use gelatin as carriers in gene delivery, and prepare the gelatin nanoparticles with particle size of 300nm. For calculating the capturing ratio, the DNA loading efficiency is nearly 80％. Size analysis and stability were performed under dynamic light scattering and TEM. DNA encapsulated in the gelatin nanoparticles could be protected from DNaseΙdegradation. In HeLa cell transfection experiments, efficiencies of commercial product LipofectaineTM 2000 and gelatin/ pEGFP-C1 nanoparticles were nearly the same. When comparing to commercial product LipofectaineTM 2000, gelatin/pEGFP-C1 nanoparticles had higher cell viability and lower cytotoxicity. After gelatin nanoparticles transfection, the egg hatchability was 80-100％. We could also detect Green fluorescence in the fourth day chicken embryo transfected by gelatin/pEGFP-C1 nanoparticles. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T07:20:12Z (GMT). No. of bitstreams: 1 ntu-97-R95548020-1.pdf: 2136748 bytes, checksum: a67999459282977ecd086e22d8494662 (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | 目錄
口試委員會審定書………………………………………………………… I 誌謝………………………………………………………………………… II 中文摘要…………………………………………………………………… III 英文摘要…………………………………………………………………… IV 目錄………………………………………………………………………… V 圖目錄……………………………………………………………………… VI 表目錄……………………………………………………………………… VIII 第一章 前言 1-1生物醫藥蛋白的生產……………………………………………… 1 1-2基因轉殖動物……………………………………………………… 2 1-3基因轉殖雞………………………………………………………… 4 1-4基因遞送系統……………………………………………………… 7 1-5高分子奈米微粒…………………………………………………… 16 1-6研究目的…………………………………………………………… 19 第二章 理論基礎 2-1明膠………………………………………………………………… 18 2-2成核理論…………………………………………………………… 22 2-3基因奈米粒子進入細胞的機制…………………………………… 26 第三章 材料與方法 3-1實驗藥品…………………………………………………………… 29 3-2實驗流程與材料製備……………………………………………… 30 3-3大量質體的萃取…………………………………………………… 31 3-4明膠奈米微粒的製備……………………………………………… 31 3-5測量粒徑以及界面電位…………………………………………… 31 3-6明膠/質體pEGFP-C1(4.7kb)奈米微粒的電泳分析……………… 31 3-7穿透式電子顯微鏡(TEM)分析…………………………………… 32 3-8質體pEGFP-C1(4.7kb)的包覆率 ………………………………… 32 3-9明膠/質體pEGFP-C1(4.7kb)奈米微粒和DNAse作用後的電泳分析 32 3-10細胞活性的WST-1測試………………………………………… 33 3-11細胞毒性LDH測試……………………………………………… 34 3-12體外轉染細胞實驗………………………………………………… 34 3-13動物實驗之體內轉染……………………………………………… 35 3-14冷凍切片觀察……………………………………………………… 35 第四章 結果與討論 4-1明膠奈米微粒的製備結果………………………………………… 36 4-1-1明膠奈米微粒的製造方式…………………………………… 36 4-1-2明膠奈米微粒粒徑分析結果………………………………… 36 4-1-3明膠奈米微粒界面電位分析結果…………………………… 42 4-2明膠/質體pEGFP-C1奈米微粒的製備結果……………………… 43 4-2-1明膠/質體pEGFP-C1奈米微粒的製備……………………… 43 4-2-2明膠/質體pEGFP-C1奈米微粒的包覆率測試……………… 45 4-2-3明膠/質體pEGFP-C1奈米微粒粒徑分析結果……………… 46 4-2-4明膠/質體pEGFP-C1奈米微粒界面電位分析結果………… 50 4-2-5明膠/質體pEGFP-C1奈米微粒抗DNase-Ι作用的電泳檢定分析 52 4-3明膠/質體pEGFP-C1奈米微粒生物相容性測試………………… 54 4-4明膠/質體pEGFP-C1奈米微粒體外螢光表現…………………… 56 4-5明膠/質體pEGFP-C1奈米微粒的體外轉染應用………………… 60 4-5-1注射雞胚方式的確認…………………………………………… 60 4-5-2 明膠/質體pEGFP-C1奈米微粒體內轉染結果……………… 63 第五章 結論……………………………………………………………… 66 參考文獻 …………………………………………………………………… 67 | |
| dc.language.iso | zh-TW | |
| dc.subject | 明膠奈米微粒、基因轉殖雞 | zh_TW |
| dc.subject | gelatin nanoparticles | en |
| dc.subject | transgenic chickens | en |
| dc.title | 明膠/DNA 奈米微粒於基因轉殖雞之應用 | zh_TW |
| dc.title | Gelatin Nanoparticles as DNA Carriers for Transgenic Chicken Application | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鄭登貴,曾厚,沙維塔(Savitha Sivasubramanian) | |
| dc.subject.keyword | 明膠奈米微粒、基因轉殖雞, | zh_TW |
| dc.subject.keyword | gelatin nanoparticles, transgenic chickens, | en |
| dc.relation.page | 68 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2008-07-25 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
| 顯示於系所單位： | 醫學工程學研究所 | |
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