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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 王愛玉 | |
dc.contributor.author | Hau-Chern Jan | en |
dc.contributor.author | 詹皓程 | zh_TW |
dc.date.accessioned | 2021-06-13T06:42:13Z | - |
dc.date.available | 2006-08-01 | |
dc.date.copyright | 2005-08-01 | |
dc.date.issued | 2005 | |
dc.date.submitted | 2005-07-31 | |
dc.identifier.citation | Averboukh L, Douglas SA, Zhao S, Lowe K, Maher J, Pardee AB (1996) Better gel resolution and longer cDNAs increase the precision of differential display. Biotechniques 20: 918-921
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/35146 | - |
dc.description.abstract | 利用基因差異表現分析法 (DD-PCR) 比較生長中綠竹筍之各組織間基因表現情形。本實驗共獲得 46 個具差異性表現之 DNA 片段,經反向北方分析 (reverse northern analysis) 結果顯示,僅 12 個 DNA 片段具有表現之差異性。其中 AA23100b-2 基因片段經定序與 BLAST 比對分析,預測為 homeoboxes 基因。為選殖出 AA23100b-2 基因全長,進一步以 AA23100b-2 基因片段作為探針,篩選綠竹筍 cDNA 庫。選殖得到之 cDNA 與水稻 putative homeodomain leucine zipper protein 高度同源,並命名為 BoHDZip,其 DNA 序列總長度為1869 bp,但仍缺少基因之 5’端序列。推衍之胺基酸序列含有 homeobox 蛋白質之 START domain 保守性胺基酸。因此推測 BoHDZip 屬於植物 homeobox 蛋白質的 HD-GL2 家族。 | zh_TW |
dc.description.abstract | Differential display-PCR (DD-PCR) was employed to compare the gene expression profiles among diverse tissues of growing bamboo shoot Bambusa oldhamii. Forty-six differentially expressed fragments were isolated, reamplified, and cloned. Reverse northern analysis showed that only twelve clones exhibited differential expression in patterns in bamboo shoot. The identity of the clone AA23100b-2 was predicted to be homeoboxes gene as revealed by sequencing analysis and BLAST analysis. To clone the full-length cDNA for AA23100b-2, a cDNA library from bamboo shoots was screened by using AA23100b-2 cDNA as probe. A cDNA whose sequence was highly homologous to the homeodomain leucine zipper protein of rice was obtained. This cDNA, designated BoHDZip was 1869bp in length, but still lacted the 5’-end region. The deduced amino acid sequence contained the conserved START domain of homeobox proteins. It might belong to the HD-GL2 family of homeobox protein in plant. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T06:42:13Z (GMT). No. of bitstreams: 1 ntu-94-R92b47203-1.pdf: 1861211 bytes, checksum: a1014242f7f895c713ae1ee700d09669 (MD5) Previous issue date: 2005 | en |
dc.description.tableofcontents | 目錄
英文摘要……………………………………………………………………………Ⅰ 中文摘要……………………………………………………………………………Ⅱ 縮寫表…………………………………………………………………..………..…Ⅲ 第一章 前言……………………………………………………….….....………….1 第一節 植物之生長發育……………………………………….……….………..1 1.1 概述…………………………………………….………….……………….1 1.2 生長受基因調控…….…………………………………….……………….2 1.3 基因表現受糖、賀爾蒙及環境之影響………………….……………….3 1.4 組織特異性基因之轉錄調控…………………………….……………….5 第二節 本論文之研究材料與方法之介紹…………………….………………..7 2.1 綠竹生長發育之特徵…………………………………….……….…....…7 2.2 利用 DD-PCR 法探討影響綠竹生長相關的基因…….………………...8 第三節 本論文之研究動機…………………………………….……….………10 第二章 材料與方法……………………………………………….….……………11 材料………………………………………………………………..……………..11 藥品………………………………………………………………….…………...11 儀器………………………………………………………………….…………...11 方法………………………………………………………………….……………12 1. 綠竹不同組織部位總RNA之抽取與檢定 1.1 總 RNA 之抽取…………….………….……………….……….…...…...12 1.2 RNA 甲醛瓊脂糖膠體電泳分析……………………….……….……….13 2. 差異性表現分析法 2.1 RNA樣品之準備……………………………….……………….………..14 2.2 cDNA之合成…………………………………….……………….………14 2.3 聚合酶連鎖反應………………………………….……………….……....15 2.4 變性聚丙烯醯胺膠體電泳……………………….………………….……15 3. 差異性表現基因之選殖與分析 3.1建構差異性基因片段之質體………………………………………….…..16 3.2 反向北方雜合分析 (reverse northern analysis)…….……………….…….22 3.3 北方雜合分析 (northern blot analysis)…………………………………....25 4. 綠竹 cDNA 庫之篩選 4.1 噬菌體效價測定…………………………………………..……………....27 4.2 cDNA之塗佈與溶菌斑之轉印…………………………..………….…....27 4.3 菌落尼龍膜雜合反應及漂洗……………………………..…………….…28 4.4 正反應溶菌斑的單離純化…………………………….….………….……29 4.5 噬菌體胞內切除 (in vivo excision)………………………..…………….29 4.6 南方氏轉印法 (southern blot analysis)…………………….….………....30 5. 其它重要的實驗 5.1 DNA 瓊脂糖膠體純化………….……………………………….………30 第三章 結果與討論……………………….…………………………………….....32 第一部分 以 DD-PCR 分析綠竹筍轉錄體(transcriptome) 第一節 綠竹不同生長時期基因之差異性表現.…………………….…………32 第二節 確認 DD-PCR基因選殖到的差異性表現….………………………….37 第三節 差異性基因於不同組織及生長時期之轉錄表現.………….…………40 第二部分 利用目標基因片段篩選綠竹筍 cDNA 庫 第一節 選殖具差異性表現目標基因之全長…………….………………….…42 第三部分 AA23100b-2選殖之Homeobox cDNA 序列分析 第一節 選殖之Homeobox cDNA 序列分析…………….………….………….44 第二節 BoHDZip 胺基酸序列之分析…………………….………….………..48 第四章 結論與未來研究方向……………………………….……………………53 第一節 結論………………………………………………….…………………53 第二節 未來方向…………………………………………….…….……………54 參考資料………………………………………………………….………………..56 | |
dc.language.iso | zh-TW | |
dc.title | 以Differential display-PCR法選殖綠竹中不同生長時期具差異性表現之基因 | zh_TW |
dc.title | Cloning of differentially expressed genes in different growth stages of green bamboo Bambusa oldhamii by differential display-PCR | en |
dc.type | Thesis | |
dc.date.schoolyear | 93-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 宋賢一,蔣啟玲,張珍田 | |
dc.subject.keyword | 綠竹,基因 差異性表現分析法, | zh_TW |
dc.subject.keyword | Green bamboo,Differential display-PCR,DD-PCR,HD-GL2 family, | en |
dc.relation.page | 60 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2005-07-31 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
顯示於系所單位: | 微生物學科所 |
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