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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 朱善德(Sin-Tak Chu) | |
| dc.contributor.author | Yi-Chieh Chiu | en |
| dc.contributor.author | 邱奕傑 | zh_TW |
| dc.date.accessioned | 2021-06-08T04:43:38Z | - |
| dc.date.copyright | 2009-08-11 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-08-04 | |
| dc.identifier.citation | Christiane Nusslein-Volhard and Ralf Dahm(2005) “Zebrafish”
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23134 | - |
| dc.description.abstract | 睪丸專一性絲胺酸/蘇胺酸磷酸酶是一個新興的磷酸酶家族，這個家族的蛋白質被認為只有睪丸表現專一性。此一蛋白質家族的成員包括TSSK1、TSSK2、SSTK、TSSK3及TSSK5，其中TSSK1、TSSK2、SSTK的功能可能與精子的發育有關，而TSSK3的功能至目前為止並不清楚。利用NCBI序列比對的結果，發現斑馬魚有一個蛋白質胺基酸序列與小鼠TSSK3具有44%的相似度，稱為zTSSK。本實驗的目的是希望藉由了解斑馬魚中zTSSK的功能，以便可以探討哺乳類TSSK3可能的生理功能，因此我們先利用斑馬魚為生物模式進行組織分布分析，在RT-PCR的結果中發現除了睪丸外，在大腦、腎臟等器官也可以看到zTSSK的表現，同時在早期胚胎發育從5hpf時期此一基因開始有表現，至72hpf時期則下降許多。利用全覆式定點雜合技術發現zTSSK在斑馬魚24、48、72 hpf的發育階段主要表現在中腦與後腦。以morpholino抑制zTSSK基因的表現則會造成胚胎的死亡，顯示在早期發育的過程中zTSSK可能扮演一個重要的角色。利用靈敏度較高的即時定量PCR發現與斑馬魚zTSSK相似的哺乳類TSSK3亦表現在小鼠的大腦及小腦組織中，而老鼠TSSK3基因的表現是在胚胎發育至14.5天TSSK3開始表現，至出生後達一個穩定值，但成熟之後表現量會下降5倍左右，此一現象和斑馬魚是相似的。因此在斑馬魚及老鼠兩個模式生物下皆可觀察到TSSK早期發育在腦部表現的情形，推測不論是zTSSK或是TSSK3，對於生物體腦的生理功能應具有一定的重要性。 | zh_TW |
| dc.description.abstract | SSK (testis specific serine/threonine kinase) family is a novel serine/threonine kinase family. The members of this family are specifically expressed in testis. Based on previous reports, TSSK1、TSSK2、and SSTK may be involved in spermatogenesis. However, the significance of TSSK3 is unclear. Using NCBI BLAST survey, we found a protein in zebrafish, namely zTSSK, which has 44% identity to TSSK3. Since zebrafish is a good animal model for studying protein function, it is helpful to use the model to elucidate the biological function of TSSK3 in mice. We demonstrate the expression of zTSSK in the tissues of brain, kidney, liver, and testis. Using whole-mount in situ hybridization technique, zTSSK has been found mainly in the midbrain and hindbrain of zebrafish at the stages of 24hpf、48hpf、and 72hpf, but decline at 72hpf stage. Knockdown of zTSSK gene by morpholino caused the death of fish embryo brfore 24hpf stage. It suggests that zTSSK play an important role during embryo development. We also found the expression of TSSK3 in mouse cerebrum and cerebellum by quantitative real-time PCR. During mouse embryonic development, TSSK3 expression begins at E14.5, reaching a maximum level before birth and remains at a constant level postnatally, but decreases in mature mice. The expression of zTSSK and TSSK3 in brain during early embryonic development suggests that both proteins play a physiological role in brain development. However, their biological roles in mice remain to be elucidated. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T04:43:38Z (GMT). No. of bitstreams: 1 ntu-98-R96b46024-1.pdf: 1074324 bytes, checksum: 8643c8f7fa5a4db7fdd69129e056362a (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | 摘要 i
Abstract ii 縮寫表 iii 目錄 v 第一章、序論 1 睪丸的介紹 1 磷酸化作用 1 睪丸專一性絲胺酸磷酸酶(Testis Specific Serine/Threonine Kinase) 3 以斑馬魚作為模式動物 5 研究動機及策略 6 第二章、實驗材料與方法 7 一、材料 7 二、實驗方法 7 1、動物組織樣本收集 7 (1.1)斑馬魚組織樣本收集 7 (1.2)斑馬魚胚胎樣本收集 7 (1.3)小白鼠組織樣本收集 8 (1.4)小白鼠胚胎樣本收集 8 2、RNA萃取 8 3、RQ1 DNase處理 9 4、反轉錄反應(Reverse transcription) 9 5、聚合酶連鎖反應(Polymerase Chain Reaction，PCR) 10 6、洋菜膠電泳 11 7、即時定量聚合酶反應(Quantitative Real-Time PCR) 12 8、核酸限制酶之切割 13 9、重組質體之建構 13 (9.1)插入片段的製備 14 (9.2)DNA模版之純化 14 (9.3)核酸限制酶切割 15 (9.4)DNA接合反應 15 (9.5)細菌轉形反應(transformation) 15 10、少量質體抽取 16 11、反義式探針(anti-sense probe)製備 17 12、全覆式定點雜合技術(whole-mount in situ hybridization) 18 13、製備Capped-RNA 21 14、以顯微注射方式注射morpholino至胚胎 21 15、Phenol/chloroform純化核酸 22 16、MO注射量的計算 22 第三章、結果 24 第一節、zTSSK基因在斑馬魚器官的基因表現 24 第二節、zTSSK在斑馬魚早期胚胎發育的表現 24 第三節、Morpholino(MO)抑制zTSSK早期胚胎發育的表現 25 第四節、TSSK3在小白鼠器官的基因表現 26 第五節、TSSK3在小白鼠胚胎發育的表現 26 第四章、討論 27 第五章、結果圖表 30 圖一、zTSSK在成熟斑馬魚組織的分布 30 圖二、zTSSK在班馬魚早期胚胎發育不同時期的表現 31 圖三、以全覆式原位雜合分析zTSSK在不同胚胎發育期表現位置 33 圖四、本實驗所用到的重組載體以及MO結合的位置 34 圖五、以zTSSK-GFP測試MO的專一性 35 圖六、抑制zTSSK在早期胚胎發育的表現導致胚胎的死亡 36 圖七、TSSK3 mRNA在老鼠組織的表現 37 圖八、TSSK3 mRNA在老鼠胚胎發育時期的表現 38 參考文獻 39 附圖一、睪丸的構造 43 附圖二、pGEMT-easy載體圖 44 附圖三、pCS2-GFP載體圖 45 | |
| dc.language.iso | zh-TW | |
| dc.subject | 磷酸酶 | zh_TW |
| dc.subject | 睪丸 | zh_TW |
| dc.subject | kinase | en |
| dc.subject | testis | en |
| dc.title | 探討睪丸專一性絲胺酸磷酸酶-TSSK特性之研究 | zh_TW |
| dc.title | Characterization of testis specific serine/threonine kinase | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 余榮熾(Lung-Chih Yu),張?仁(Ching-Jin Chang),管永恕(Yung-Shu Kuan) | |
| dc.subject.keyword | 睪丸,磷酸酶, | zh_TW |
| dc.subject.keyword | testis,kinase, | en |
| dc.relation.page | 45 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2009-08-04 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科學研究所 | zh_TW |
| 顯示於系所單位： | 生化科學研究所 | |
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