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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 齊肖琪(Shau-Chi Chi) | |
| dc.contributor.author | Li-Ling Huang | en |
| dc.contributor.author | 黃麗玲 | zh_TW |
| dc.date.accessioned | 2021-06-13T07:13:30Z | - |
| dc.date.available | 2008-07-28 | |
| dc.date.copyright | 2005-07-28 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-07-26 | |
| dc.identifier.citation | 丁詩同及鄭登貴（2003） 脂肪細胞學研究技術專輯 國立台灣大學畜產學系
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/35849 | - |
| dc.description.abstract | 神經壞死症病毒（Nervous Necrosis Virus, NNV ）在感染石斑魚苗後，會引起急性發病及大量死亡。本研究目的在找出因NNV感染而表現量有明顯差異的寄主基因，並觀察這些基因表現量在感染病毒後的時序變化。採收NNV感染前後的石斑魚苗的mRNA，用抑制性雜合扣除法（Suppression Subtractive Hybridization, SSH），以及北方雜合法（Northern analysis）篩選出四株在感染NNV前後有差異表現量的寄主基因，這些基因序列經由Blast-n比對後，由正向扣除所篩選的FB2序列是alpha-microglobulin/HI-30 precursor，FB9序列是glyceraldehyde 3-phosphate dehydrogenase（G3PDH），在反向扣除所篩選到的R1H1序列是beta globin，R2H12是liver-basic fatty acid binding protein（Lb-FABP）。比較這四個基因於NNV感染後不同天數的表現量，發現代號為FB2及FB9的兩個基因，在感染末期的瀕死魚體內表現量激增；而代號為R1H1及R2H12的兩個基因，則在感染後表現量驟減。 | zh_TW |
| dc.description.abstract | Nervous Necrosis Virus（NNV）can induce acute infection and mass mortality of grouper fry and larva. The aim of this study is to detect and analysis of the differentially expressed grouper genes after NNV infection. By suppression subtractive hybridization (SSH) and Northern analysis, two cDNA sequences（FB2, FB9） were selected by forward subtraction and two cDNA sequences（R1H1, R2H12）were selected by reverse subtraction. These four cDNA sequences were analyzed by Blast-n program, and the results revealed that FB2 is similar to the gene of alpha-microglobulin/HI-30 precursor, FB9 is similar to the gene of glyceraldehyde 3-phosphate dehydrogenase (G3PDH), R1H1 is similar to the gene of beta globin, and R2H12 is similar to the gene of liver-basic fatty acid binding protein (Lb-FABP). The expression of either FB2 and FB9 increased significantly in the moribund fish 9 days after NNV infection. On the other hand, the expression of R1H1 and R2H12 decreased drastically in the fish showing clinical syndrome of NNV disease in the early stage of NNV infection. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T07:13:30Z (GMT). No. of bitstreams: 1 ntu-94-R92b41006-1.pdf: 1187078 bytes, checksum: 2309847a6fe1b55dea0889e39b3499b6 (MD5) Previous issue date: 2005 | en |
| dc.description.tableofcontents | 目 錄
目錄 Ⅰ 圖表目次 Ⅳ 中文摘要 Ⅴ 英文摘要 Ⅵ 第一章 緒論 1. 病毒性神經壞死症之病史及病癥 1 2. 病毒性神經壞死症之分子生物特性 1 3. 神經壞死症病毒之分類 2 4. 神經壞死症病毒於宿主體內之分佈 3 5. 對神經壞死症病毒具感受性之細胞株 4 6. 神經壞死症病毒的診斷方法 5 7. 病毒性神經壞死症疫情的控制方法 6 8. 病毒感染細胞及宿主個體後的反應 7 9. 抑制性雜合扣除法之介紹 8 10. 實驗目的 11 第二章 材料與方法 1. In vivo寄主RNA之收集 12 2. Total RNA之萃取 13 3. 抑制性扣除雜合法（SSH） 13 3.1 mRNA之萃取 14 3.2 第一股cDNA之合成 15 3.3 第二股cDNA之合成 16 3.4 Rsa I限制酶截切 17 3.5 Adaptor的接合 18 3.6 Adaptor接合效率分析 19 3.7 第一次雜合 20 3.8 第二次雜合 21 3.9 第一次PCR 21 3.10 第二次PCR 22 3.11 扣除效率分析 23 4. T/A cloning 4.1 pGEM○R-T easy Vector接合反應 24 4.2以熱休克法轉形勝任細胞 25 4.3菌落挑選 25 5. 差異性基因篩選（Differential screening） 5.1 雜交膜之製備 26 5.2 核酸探針之製備 27 5.3 雜合反應 28 5.4 結果判讀 28 6. 北方吸漬法（Northern analysis） 6.1 質體DNA之抽取 29 6.2 雜交膜之製備 30 6.3 核酸探針之製備 31 6.4 雜合反應 32 6.5 結果判讀 33 7. 及時定量聚合酶連鎖反應（Real-Time Quantitative PCR） 7.1 反轉錄反應 33 7.2 進行Real-Time Quantitative PCR反應 34 第三章 結果 1. 利用抑制性扣除雜合法建立NNV感染石斑魚苗前後之基因庫 1.1 石斑魚苗感染NNV後的病程及累積死亡率 36 1.2 萃取石斑魚苗RNA 36 1.3 利用抑制性扣除雜合法得到之cDNA基因庫 37 1.4 基因庫序列分析 38 2. 以北方雜合法確認基因表現差異 38 3. 以Real-Time PCR分析基因表現 39 4. 有差異性表現基因之比對結果 39 第四章 討論 1. 以抑制性扣除雜合法建立基因庫 41 2. 具有差異性表現的基因 42 第五章 參考文獻 47 | |
| dc.language.iso | zh-TW | |
| dc.title | 比較石斑魚苗在感染神經壞死症病毒前後寄主基因表現之差異 | zh_TW |
| dc.title | Comparisons on differentially expressed genes in grouper（Epinephelus coioides）fry with nervous necrosis virus infection | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 羅竹芳,郭欽明,張朴性 | |
| dc.subject.keyword | 抑制性雜合扣除法,神經壞死症病毒,北方雜合法,石斑魚苗, | zh_TW |
| dc.subject.keyword | Suppression Subtractive Hybridization, SSH,Nervous Necrosis Virus, NNV,Northern analysis,grouper fry, | en |
| dc.relation.page | 73 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2005-07-26 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 動物學研究研究所 | zh_TW |
| 顯示於系所單位： | 動物學研究所 | |
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