請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/47204完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 吳信志(Shinn-Chih Wu) | |
| dc.contributor.author | Yun-Yi Ma | en |
| dc.contributor.author | 馬耘逸 | zh_TW |
| dc.date.accessioned | 2021-06-15T05:50:45Z | - |
| dc.date.available | 2012-08-20 | |
| dc.date.copyright | 2010-08-20 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-08-18 | |
| dc.identifier.citation | 65
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/47204 | - |
| dc.description.abstract | 3
摘要 本研究主旨在探討小鼠原核胚經雙基因共同注射後，於體外培養至二細 胞時期再進行胚移置對產製攜帶雙基因轉殖小鼠效率之影響。轉殖基因之構築 係以乳腺專一表現之牛 | zh_TW |
| dc.description.abstract | 74
英文摘要 Generation of Transgenic Mice Harboring Human Pulmonary Surfactant-related Protein B/C Genes Yun-Yi Ma Abstract The purpose of this study was to investigate the effect of advanced culture to two-cell stage prior to co-injected mouse eggs on pups born and transgene integration efficiency by pronuclear gene microinjection. Transgenes were constructed by using a mature surfactant protein B (mSP-B) and C (mSP-C) cDNA fragment equipped with bovine | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T05:50:45Z (GMT). No. of bitstreams: 1 ntu-99-R97626012-1.pdf: 4287112 bytes, checksum: 2ac646f8f862190852cd263b91e73661 (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 目錄
頁次 目錄 …………………………………………………………………………………… II 表次 …………………………………………………………………………………… V 圖次 …………………………………………………………………………………… IV 摘要 …………………………………………………………………… 3 緒言 …………………………………………………………………… 5 第一章、 文獻檢討 ………………………………………………………… 7 第一節 基因轉殖動物 …………………………………………………… 7 (一) 基因轉殖定義與技術原理……………………………………… 7 (二) 產製基因轉殖動物之方法……………………………………… 7 (三) 基因轉殖動物之研究進展……………………………………… 12 (四) 產製基因轉殖動物產製效率之改善 ………………………… 12 (五) 基因轉殖動物之應用…………………………………………… 14 第二節 肺表面張力劑(surfactant) ………………………………… 24 (一) (二) 肺表面張力劑之簡介…………………………………………… B 型肺臟表面張力劑相關蛋白質之功能與特性 ……………… 24 26 (三) (四) C 型肺臟表面張力劑相關蛋白質之功能與特性 ……………… 新生兒呼吸窘迫症之簡介……………………………………… 28 33 (五) 醫療用疏水性肺臟表面張力劑蛋白質之產製現況…………… 36 第二章、 人類肺表面張力蛋白質 B/C 基因轉殖小鼠之產製與初步分析 41 第ㄧ節 前言 …………………………………………………………… 41 口試委員會審定書………………………………………………………………… I Index of Tables ……………………………………………………………… VI Index of Figures ……………………………………………………………… VII II Index of Figures ………………………………………………………………… I 第二節 材料與方法 …………………………………………………… 42 (ㄧ) 轉殖基因之製備 ……………………………………………… 42 (二) 試驗動物之飼養及超級排卵處理 …………………………… 42 (三) (四) (五) (六) 小鼠原核胚之基因顯微注射…………………………………… 基因轉殖小鼠之 PCR 初步分析………………………………… 各仔小鼠基因組 DNA 之 Southern-blot 與 Slot-blot 分析 轉殖基因在基因轉殖小鼠性腺中之傳承……………………… 43 44 45 46 第三節 結果與討論 …………………………………………………… 49 (ㄧ) (二) (三) (四) 基因顯微注射之鼠胚經培養至二細胞後移置於受胚母鼠後胚之 存活率及其基因轉殖效率 …………………………………… 基因轉殖小鼠之 PCR 初步分析的結果………………………… 基因轉殖小鼠外源基因疊接型態之 Southern-blot 與 Slot-blot 分析 …………………………………………………………… 外源基因在基因轉殖小鼠性腺中之傳承 …………………… 49 49 50 52 第三章、 外源基因於基因轉殖小鼠乳腺中表現之分析 ……………… 60 第ㄧ節 前言 …………………………………………………………… 60 第二節 材料與方法 …………………………………………………… 60 (一) 源自基因轉殖小鼠之乳汁中蛋白質之 Western-blot 分析… 60 第三節 結果與討論 …………………………………………………… 61 (一) 外源基因在基因轉殖小鼠乳腺中之表現分析 ……………… 61 結論 參考文獻 ………………………………………………………………… ………………………………………………………………… 64 65 英文摘要 ………………………………………………………………… 74 III Index of Figures ………………………………………………………………… I 表次 表 1 . 滿足全球生技藥品市場需求所需的基因轉殖動物數目…………… 22 表 2 . 基因轉殖動物所生產藥物的經濟效益……………………………… 22 表 3 . 自乳汁中所生產的醫藥用蛋白質…………………………………… 23 表 4 . 成 熟 的 肺 表 面 張 力 素 組 成 …………………………………… 23 表 5 . 目前於醫療用之外源性肺表面張力劑種類………………………… 40 表 6 . 轉殖基因 PCR 分析所使用之寡核苷酸引子對……………………… 54 表 7 . 以雙基因共同顯微注入小鼠胚原核後於原核時期及續培養至 二細胞期進行胚移置對轉殖基因嵌合效率之影響………………… 55 表 8 . 基因轉殖小鼠之外源基因性腺傳承分析結果 ……………………… 59 頁次 IV Index of Figures ………………………………………………………………… I 圖次 頁次 圖 1. 產製基因轉殖動物之方法………………………………………… 11 圖 2. 圖 3. 圖 4. 圖 5. 圖 6. 圖 7. 圖 8 圖 9 圖 10 圖 11 肺臟表面張力劑於第二型肺泡細胞內之合成、加工及再利用路徑 人類 B 型與 C 型肺臟表面張力劑相關蛋白質(SP-B and SP-C) 之訊 息核糖核酸 (mRNA) 與蛋白質之轉錄與轉譯作用…………… 人類 B 型與 C 型肺臟表面張力劑相關蛋白質之蛋白質結構……… 新生兒呼吸窘迫症之致病機轉…………………………………… αLA-CNHIS-mHSPB/C 轉殖基因之構築…………………………… 小鼠原核胚之基因顯微注射……………………………………… αLA-CNHIS-mHSPB 基因轉殖小鼠之 PCR 分析結果…………… αLA-CNHIS-mSPB /C 基因轉殖小鼠之 Southern-blot 雜合分析… αLA-CNHIS-mHSPB/C 在基因轉殖小鼠基因組中嵌插之套數，經 slot-blot 雜合分析之結果…………………………………………… αLA-CNHIS-mHSPB/C 基因轉殖小鼠 (No.7, 10♀)西方點墨法之乳樣 分析 ……………………………………………………………… 人 26 31 32 40 47 48 56 57 58 63 V I Index of Tables Ta b l e 1 . Number of transgenic animal species to satisfied the global market of pharmaceutical protein ………………………………………… 22 Ta b l e 2 . Economic benefits of pharmaceutical proteins produced by transgenic animals……………………………………… 22 Ta b l e 3 . Some of the pharmaceutical proteins under study produced in milk…………………………………………………………………… 23 Ta b l e 4 . Composition of mature surfactant…………………………………… 23 Ta b l e 5 . Clinical exogenous surfactant drugs used to treat lung diseases of surfactant deficiency/dysfunction................... 40 Ta b l e 6 . Oilgonucleotide primer pairs used for PCR analysis in transgenesis process…………………………………………………… 54 Ta b l e 7 . The effects of advanced culture to two-cell stage prior to transgenes co-injected mouse eggs on pregnant, pups born and transgne integration rate……………………………… 55 Ta b l e 8 . Germine transmission of transgenic mouse assessed by analysis of genomic DNA samples from F1 progenies had been generated by breeding each of transgenic mice with the nontransgenic mice……… 59 VI Index of Figures ………………………………………………………………… I 2 Index of Figures Figure 1. Different methods to generate transgenic animals.……………………… 11 Figure 2. Figure 3. Figure 4. Figure 5. Figure 6. Figure 7. Figure 8. Figure 9. Figure 10. Figure 11. Biosynthesis、processing and recycling of pulmonary surfactant in the type II pneumocytes. …………………………………………………… Transcription and translation of the human pulmonary surfactant-associated protein B and C mRNA. ………………………… The protein structure of human surfactant-associated protein B(SP-B) and C(SP-C) .………………………………………………………… The pathological mechanism of neonatal respiratory distress syndrome. Plasmid construction of αLA-CNHIS-mHSPB/C exogenic gene.……… Gene microinjected into pronucleus of mouse embryo……………… Transgenic mice carrying with αLA-CNHIS-mHSPB/C detected by PCR screening.……………………………………………………………… Southern-blot hybridization analysis of αLA-CNHIS-mSPB /C transgenic mice.……………………………………………………… The detection of transgene copy numbers in αLA-CNHIS-mHSPB/C transgenic mice by slot-blot hybridization analysis.………………… Western-blot analysis of the milk obtained from transgenic mice (No. 7, 10♀) carrying with αLA-CNHIS-mHSPB/C.…………………………… 人 26 31 32 40 47 48 56 57 58 63 | |
| dc.language.iso | zh-TW | |
| dc.subject | C型蛋白質 | zh_TW |
| dc.subject | 乳汁 | zh_TW |
| dc.subject | 雙基因共同注射 | zh_TW |
| dc.subject | B 型肺臟表面張力蛋白質 | zh_TW |
| dc.subject | 基因轉殖小鼠 | zh_TW |
| dc.subject | microinjection | en |
| dc.subject | transgenic mice | en |
| dc.subject | surfactant protein B | en |
| dc.subject | surfactant protein C | en |
| dc.subject | milk | en |
| dc.title | 產製人類肺表面張力蛋白質 B/C 基因轉殖小鼠之研究 | zh_TW |
| dc.title | Generation of Transgenic Mice Harboring Human
Pulmonary Surfactant-related Protein B/C Genes | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 黃木秋(Mu-Chiou Huang),陳全木(Chuan-Mu Chen),宋麗英(Li-Ying Sung) | |
| dc.subject.keyword | 雙基因共同注射,基因轉殖小鼠,B 型肺臟表面張力蛋白質,C型蛋白質,乳汁, | zh_TW |
| dc.subject.keyword | microinjection,transgenic mice,surfactant protein B,surfactant protein C,milk, | en |
| dc.relation.page | 75 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2010-08-18 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 動物科學技術學研究所 | zh_TW |
| 顯示於系所單位： | 動物科學技術學系 | |
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