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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 丁詩同(Shih-Torng Ding) | |
| dc.contributor.author | Pin Chang | en |
| dc.contributor.author | 張斌 | zh_TW |
| dc.date.accessioned | 2021-06-15T04:53:43Z | - |
| dc.date.available | 2012-06-12 | |
| dc.date.copyright | 2010-08-05 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-07-30 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46093 | - |
| dc.description.abstract | 母豬繁殖性能是養豬產業生產效率之重要因子，當中又以窩仔數為最重要之經濟繁殖性狀，若能改進窩仔數將能提升養豬產業整體之經濟效益。近年來，隨著分子生物技術之發展，藉由候選基因分析方法（candidate gene approach）找尋分子遺傳標記，並應用於標記輔助選拔（marker-assisted selection, MAS），將可大幅改進母豬繁殖性狀之選拔效應。藍瑞斯品種母豬為臺灣育種策略中的重要豬種，但至今仍未出現能夠顯著影響純種藍瑞斯母豬窩仔數的分子遺傳標記。
豬桑椹胚至囊胚時期的發育過程，為埋殖前胚成功著床以及胎體成功發育之關鍵時期。因此，本論文藉由桑椹胚發育至早期囊胚階段中，mRNA 表現量顯著增加或降低之基因作為候選基因，並進一步找尋其調控區域之單一核苷酸多型性 （single nucleotide polymorphism, SNP）分子遺傳標記，同時建立高通量（high-throughput）SNP基因型分析系統平台，利用高通量基因型分析方法不僅降低每一個SNP位點分析的成本，並可以同時大量且快速的鑑定 SNP 基因型作為瞭解其與窩仔數之間的關係，期能篩選出針對藍瑞斯母豬窩仔數高低產的標記，以協助遺傳育種改良。 來自四間種豬場且無相關遺傳背景之純種藍瑞斯母豬共30頭定序後發現，在 19 個候選基因裡，共有 78 個 SNP發生於調控區域、11個 SNP 位於 5’ 端未轉譯區域（untranslated region, UTR），有 1個發生於轉譯區域及2個位於內插子（intron）的 SNP，合計共有 92 個新開發之 SNP。進一步利用 SNP 基因定型系統鑑定 460 頭純種藍瑞斯母豬之 SNP 基因型，同時分析各 SNP 基因型對於母豬不同胎次之出生仔豬總頭數（total number of pigs born, TNB）與存活仔豬頭數（number born alive, NBA）之差異。結果發現，位於 LIM and SH3 protein 1（LASP-1）、activatin signal cointrgrator 1 complex subunit 3（ASCC3）、ataxia-telangiectasia mutated （ATM）、plasma platelet-activating factor acetylhydrolase（plasma PAFAH）、serum amyloid A （SAA）、與visfatin等基因調控區域之SNP分子標記，其基因型與藍瑞斯品種母豬之TNB 及 NBA 有顯著關係存在（P < 0.05），說明了這些調控區域的 SNP 可能與控制基因表現有密切關係存在。 本研究新開發之SNP標記，發現為影響純種藍瑞斯母豬窩仔數的分子遺傳標記，未來若能取得更多不同豬場繁殖的藍瑞斯母豬樣本數及生產紀錄，將能更進一步確認這些 SNP 在遺傳育種改良上之價值，並將有機會藉由標記輔助選拔應用於傳統育種選拔上，以改進傳統育種對於母豬繁殖性狀之選拔效率，並成功增進畜牧產業於臺灣農業之產值。 | zh_TW |
| dc.description.abstract | Reproduction performance of sows is one of the most important factors for pig industry, which is the mainstream of livestock production in Taiwan. Improvement of the litter size, a critical reproduction trait, by traditional genetic selection methods generates limited effect, due to the low heritability and sex-limited trait. Using single nucleotide polymorphism (SNP) molecular markers for a marker-assisted selection program through candidate gene approach may improve reproductive traits. The purpose of this study was to investigate the genetic polymorphisms in candidate genes and their association with reproductive performance in Landrace sows, the major breed used as dam in commercial pig production in Taiwan.
Candidate genes were selected from embryonic mRNA differentially expressed between morula and blastocysts. After cloning and sequencing the promoter region of the 19 candidate genes, we found 92 novel SNPs from DNA sequences of 30 unrelated Landrace pigs. There are 78 SNPs located on regulatory region, 11 SNPs on the 5’ UTR, 2 SNPs on the intron and 1 SNP located on the CDS. Furthermore, by using the high-throughput Illumina GoldenGate® genotyping assay, we found 10 novel SNP markers significantly associated with total number born (TNB) and number born alive (NBA), including genes of activating signal co-integrator 1 complex subunit 3 (SSC.13633-B, P < 0.05), LIM and SH3 protein 1 (PLAb11012-C, P < 0.05; PLAb11012-E, P < 0.01), ataxia-telangiectasia mutated（SSC.21876-A, P < 0.05; SSC.21876-L, P < 0.01）, plasma platelet-activating factor acetylhydrolase（SSC.19691-A, P < 0.05; SSC.19691-F, P < 0.01）, serum amyloid A（SAA-Q, P < 0.05 ; SAA-AA, P < 0.05）, and Visfatin (Visfatin-A, P < 0.05) in 460 Landrace sows from two commercial breeding herds. These litter size-related SNPs are good candidate molecular markers for improving the reproductive efficiency of pigs. The present study developed an effective platform for determining the litter size-related SNPs, which might be associated with the regulatory region of differentially regulated genes from morula to blastocyst. Once validated further with large sample size and collected fully-formed records from different herds that would be applied to marker-associated selection to improve reproductive performance in Landrace sows, and increased the production value on the pig production industry. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T04:53:43Z (GMT). No. of bitstreams: 1 ntu-99-R97626013-1.pdf: 4806458 bytes, checksum: e6c781a6904d6b1bfd2a2bdd9a9ce3e2 (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 目錄 I
圖目錄 VI 表目錄 VII 中文摘要 IX 英文摘要 XI 壹、前言 1 貳、文獻檢討 3 一、臺灣養豬產業現況 3 （一）繁殖能力於臺灣養豬產業之重要性 3 （二）母豬繁殖能力之影響因子 4 1. 窩仔數對於繁殖性狀的重要性 4 2. 離乳至配種間距及胎次對窩仔數的影響 5 3. 配種月份對出生活仔頭數之影響 6 （三）基因輔助選拔方式相較傳於統育種選拔之優勢 6 （四）母豬繁殖性狀相關之候選基因 7 1. 動情素受體（Estrogen receptor, ESR） 8 2. 泌乳素受體（Prolactin receptor, PRLR） 8 3. 瘦體素（Leptin, LEP） 9 4. 白血病抑制因子（Leukemia inhibitory factor, LIF） 11 5. p53腫瘤抑制因子（p53 tumor suppression gene） 11 6. 其他影響繁殖相關性狀之 DNA 分子遺傳標記 12 二、開發與母豬窩仔數相關之單一核苷酸多型性分子遺傳標記 15 （一）DNA分子遺傳標記 15 1. DNA 遺傳標記之分類 16 2. 偵測 DNA 遺傳標記之技術 16 （二）單一核苷酸多型性 18 （三）鑑定單一核苷酸多型性之方法 19 1. 引子延伸法（Primer extension） 19 2. 雜合法（Hybridization） 21 3. 連結法（Ligation） 21 4. 酵素截切法（Enzymatic cleavage） 23 5. 利用分子量偵測其基因型 23 6. 其他鑑定基因型之方法 23 （四）高通量鑑定單一核苷酸多型性之應用 25 1. Vera Code技術 25 2. GoldenGate®檢測技術 26 3. SNPlexTM檢測技術 26 4. TaqMan®檢測技術 27 三、基因圖譜之應用 29 （一）數量性狀基因座之確認 29 （二）標記輔助選拔（Marker- assisted selection, MAS） 30 （三）豬基因組圖譜之研究 30 （四）豬數量性狀基因座 30 （五）豬窩仔數相關之候選基因 31 1. 候選基因：豬桑椹胚及早期囊胚差異性表現基因 31 1-1. 豬胚埋殖前胚發育過程 32 1-2. 建立豬胚埋殖前胚基因表現模式之研究 34 1-3. 豬桑椹胚及早期囊胚差異性表現之基因 34 2. 候選基因：脂肪代謝調節相關研究基因 37 2-1. 脂締素基因（Adiponectin） 38 2-2. 內臟脂肪素基因（Visfatin） 38 2-3. 過氧化酶體增生活化受體γ共激活因子-1α（PPARGC1A） 39 四、研究動機及目標 40 参、材料與方法 42 試驗一、候選基因調控區域之選殖 42 （一）候選基因之選擇 42 （二）選殖調控區域序列 43 （三）次選殖 43 試驗二、母豬窩仔數相關SNP基因型之開發 45 （一）試驗材料與資料蒐集 45 （二）豬基因組DNA之萃取 45 （三）聚合酶連鎖反應（PCR） 46 （四）定序分析 46 試驗三、高通量SNP基因型與母豬窩仔數之遺傳相關 48 （一）試驗材料與資料蒐集 48 （二）豬基因組DNA之抽取 48 （三）高通量分析 SNP 系統平台之建構與分析 48 1. 使用軟體進行Illumina GoldenGate引子設計 49 2. ADT軟體設計檔案格式 49 3. 樣本DNA製備 50 4. GoldenGate反應 51 5. 雜合反應 54 （四）影像偵測分析 55 （五）統計分析 55 （六）分析 SNP系統平台之建構與分析 58 肆、試驗結果 61 試驗一、候選基因調控區域之選殖 61 （一）豬基因組庫之建構 61 （二）候選基因調控與次選殖 61 試驗二、母豬窩仔數相關SNP基因型之開發 63 （一）候選基因調控區域序列分析與SNP分析 63 （二）母豬繁殖性狀相關之DNA分子遺傳標記之SNP基因型 63 （三）候選SNP之調控功能 63 試驗三、高通量SNP基因型與母豬窩仔數之遺傳相關 72 （一）利用高通量技術篩選對於繁殖性狀有差異的候選SNP 72 （二）候選基因SNP基因型與母豬窩仔數之遺傳相關 72 伍、討論 96 一、新開發之SNP數量 96 二、SNP 基因定型系統之建立及其效能 96 三、SNP 基因型與母豬窩仔數 98 （一）新開發之SNP與窩仔數相關之分子標記 98 （二）文獻中與母豬繁殖性狀相關之 DNA 分子遺傳標記 100 （三）開發藍瑞斯母豬窩仔數性狀之新穎SNP分子遺傳標記 102 四、候選基因應用於家畜育種選拔計畫 103 五、未來方向 103 陸、結論 106 參考文獻 107 附錄 126 【附錄一】候選基因之序列與引子設計 126 附表1. 選殖各候選基因調控區域之引子設計表 126 附表2. 候選基因DNA分子遺傳標記之PCR-RFLP條件 128 附表3. ASPE-PCR 反應所需之引子表 129 附表4. 線上分析設計工具軟體分析GoldenGate評分表 130 【附錄二】SNP基因型分析結果 136 附表5. 候選基因中之 SNP 的基因型頻率 136 附表6. SNP基因型對第一胎次窩仔數影響 138 附表7. SNP基因型對第二胎次窩仔數影響 144 附表8. SNP基因型對第三胎次窩仔數影響 150 附表9. SNP基因型對第四胎次窩仔數影響 156 附表10. SNP基因型對第五胎次窩仔數影響 162 附表11. SNP基因型對第二至第三胎次窩仔數影響 168 附表12. SNP基因型對第二至第四胎次窩仔數影響 174 附表13. SNP基因型對第二至第五胎次窩仔數影響 180 【附錄三】樣本採集及分析結果圖 186 附圖 1. 樣本採集地區及其平均氣溫 186 附圖 2. 基因型螢光強度分析分群及數量圖 186 | |
| dc.language.iso | zh-TW | |
| dc.subject | 高通量基因型分析 | zh_TW |
| dc.subject | 酸多型性 | zh_TW |
| dc.subject | 單一核苷 | zh_TW |
| dc.subject | Plasma platelet-activating factor acetylhydrolase 基因 | zh_TW |
| dc.subject | 窩仔數 | zh_TW |
| dc.subject | 藍瑞斯母豬 | zh_TW |
| dc.subject | serum amyloid A基因 | zh_TW |
| dc.subject | LIM and SH3 protein 1 基因 | zh_TW |
| dc.subject | Visfatin基因 | zh_TW |
| dc.subject | 差異性表現基因 | zh_TW |
| dc.subject | activatin signal cointrgrator 1 complex subunit 3基因 | zh_TW |
| dc.subject | 早期豬胚發育 | zh_TW |
| dc.subject | Ataxia-telangiectasia mutated 基因 | zh_TW |
| dc.subject | Visfatin gene | en |
| dc.subject | Landrace sows | en |
| dc.subject | Litter size | en |
| dc.subject | Single nucleotide polymorphism | en |
| dc.subject | Early embryo | en |
| dc.subject | High-throughput genotyping | en |
| dc.subject | Differentially expressed gene | en |
| dc.subject | LIM and SH3 protein 1 gene | en |
| dc.subject | activatin signal cointrgrator 1 complex subunit 3 gene | en |
| dc.subject | Ataxia-telangiectasia mutated gene | en |
| dc.subject | Plasma platelet-activating factor acetylhydrolase gene | en |
| dc.subject | serum amyloid A gene | en |
| dc.title | 利用高通量鑑定單一核苷酸多型性研發藍瑞斯母豬窩仔數相關分子標記 | zh_TW |
| dc.title | Using High-Throughput Single Nucleotide Polymorphism Genotyping to Reveal Litter Size-Related Molecular Markers in Landrace Sows | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 林恩仲(En-Chung Lin) | |
| dc.contributor.oralexamcommittee | 黃木秋,王佩華 | |
| dc.subject.keyword | 藍瑞斯母豬,窩仔數,單一核苷,酸多型性,高通量基因型分析,早期豬胚發育,差異性表現基因,LIM and SH3 protein 1 基因,activatin signal cointrgrator 1 complex subunit 3基因,Ataxia-telangiectasia mutated 基因,Plasma platelet-activating factor acetylhydrolase 基因,serum amyloid A基因,Visfatin基因, | zh_TW |
| dc.subject.keyword | Landrace sows,Litter size,Single nucleotide polymorphism,Early embryo,High-throughput genotyping,Differentially expressed gene,LIM and SH3 protein 1 gene,activatin signal cointrgrator 1 complex subunit 3 gene,Ataxia-telangiectasia mutated gene,Plasma platelet-activating factor acetylhydrolase gene,serum amyloid A gene,,Visfatin gene, | en |
| dc.relation.page | 186 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2010-07-30 | |
| 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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