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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳昭瑩 | |
dc.contributor.author | Shih-Min Wu | en |
dc.contributor.author | 吳詩敏 | zh_TW |
dc.date.accessioned | 2021-06-13T06:19:17Z | - |
dc.date.available | 2011-02-20 | |
dc.date.copyright | 2006-02-06 | |
dc.date.issued | 2006 | |
dc.date.submitted | 2006-01-25 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/34630 | - |
dc.description.abstract | 葵百合LsGRP1(Lilium cv. Star Gazer glycine-rich protein 1)cDNA,為利用抗病誘導物質水楊酸(salicylic acid,SA),以PCR-select subtraction法及differential screening篩選得到的防禦相關基因,並已推測LsGRP1在百合由水楊酸與病原菌誘導的防禦反應中為重要的因子。本研究利用北方雜合分析偵測葵百合接受不同刺激之LsGRP1的表現量,結果顯示除了水楊酸,甲基茉莉酸和胺基環丙烷羧酸也會誘導LsGRP1的表現。為得到LsGRP1訊息RNA的全長度cDNA,利用快速擴增cDNA末端法,得到LsGRP1之5’端未轉譯區,包含起始密碼前21 bp;並得到相似性序列,推測葵百合有LsGRP1同源基因。分別在葵百合基因組DNA及訊息RNA上找尋LsGRP1同源基因,於前者進行聚合酵素連鎖反應,結果發現葵百合LsGRP1基因中包含了180 bp的隱子(intron),此外並發現利用不同引子對,以基因組DNA與訊息RNA分別進行基因組與反轉錄聚合酵素連鎖反應均得到多重產物;其中得到一cDNA較LsGRP1多出208 bp的重複性序列,命名為LsGRP2。另一以3’端快速擴增cDNA末端法得到之同源基因LsGRP3,僅可轉譯出70個胺基酸。由5’端與3’快速擴增cDNA法得到比LsGRP1多39 bp重複序列的LsGRP4,其轉譯出的胺基酸序列與LsGRP1相似度為90%。本研究指出葵百合LsGRP1於基因體中存在同源基因,然此等基因是否表現或其表現情形並不清楚。 | zh_TW |
dc.description.abstract | A cDNA, named LsGRP1, has been obtained by suppression subtractive hybridization from salicylic acid (SA)-treated Lilum cv. Star Gazer plants, followed by differential screening. In the previous study, LsGRP1 was presumed to play a role in SA and pathogen-induced defense responses in lily. In this study, the expression of LsGRP1 in ‘Star Gazer’ leaves was analyzed by Northern blot analysis. The result showed that not only SA, but also methyl jasmonate and 1-aminocyclopropane-1-
carboxylic acid can induce LsGRP1 expression. Rapid amplification of cDNA ends was performed to obtain 5’-untranslated region of LsGRP1,that is 21-bp upstream of the start codon. In addition, the homologue of LsGRP1 was investigated on the genome and mRNA of ‘Star Gazer’. By the way, LsGRP1 gene was amplified from genomic DNA of ‘Star Gazer’ by polymerase chain reaction (PCR), which contains a 180-bp intron. On the other hand, multiple fragments were amplified from genomic DNA and mRNA by PCR and reverse transcription-PCR using different primer pairs. A fragment which a 208-bp repeated sequence of LsGRP1 was named LsGRP2. The second homologues of LsGRP1, named LsGRP3, only encoded 70 amino acid residues. The third homologue was 39-bp longer than LsGRP1, encoding a peptide sequence of 90% identity to that of LsGRP1. These results indicated that homologues of LsGRP1 present in the genome of Lilium cv. Star Gazer although the expression of these homologues has not yet be analyzed. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T06:19:17Z (GMT). No. of bitstreams: 1 ntu-95-R92633015-1.pdf: 1424338 bytes, checksum: 7b827930d789156e575ff4b78ef5979a (MD5) Previous issue date: 2006 | en |
dc.description.tableofcontents | 壹、中文摘要 4
貳、英文摘要 5 參、前言 6 肆、前人研究 8 一、植物細胞壁蛋白質 8 二、富含甘胺酸蛋白質 8 三、植物GRPs在生長發育中表現與調控 10 四、外在因子誘導植物GRPs的表現 10 五、植物GRPs基因家族 14 六、葵百合之LsGRP1 14 伍、材料與方法 16 一、供試植物 16 二、葵百合基因組DNA之萃取 16 三、葵百合全RNA之萃取 17 四、葵百合葉片訊息RNA之萃取 17 五、反轉錄(Reverse Transcript) 18 六、引子設計 18 七、聚合酵素連鎖反應(Polymerase chain reaction,PCR) 19 八、快速擴增cDNA末端(Rapid amplification of cDNA ends,RACE) 19 九、TA-cloning 21 十、核酸探針製備 23 十一、菌落聚合酵素連鎖反應(Colony PCR) 24 十二、以點漬雜合分析(dot blot)篩選具有LsGRP1序列之選殖株 24 十三、序列分析 25 十四、化學物質處理 26 十五、北方雜合分析 26 陸、結果 28 一、LsGRP1基因表現 28 二、5’端快速擴增cDNA末端法得到LsGRP1之5’端未轉譯區與相似序列 29 三、利用基因組聚合酵素連鎖反應探討LsGRP1之同源基因 30 四、利用反轉錄聚合酵素連鎖反應探討LsGRP1之同源基因 31 五、引子293增幅LsGRP1較為專一 32 六、3’ 端快速擴增cDNA末端法得到LsGRP1相似序列 32 六、LsGRP4 33 七、 選殖策略 35 柒、討論 36 捌、參考文獻 41 玖、圖表集 46 表一、引子對序列 46 圖一、LsGRP1引子位置 48 圖二、甲基茉莉酸處理葵百合葉片,處理葉全RNA之北方雜合分析 49 圖三、甲基茉莉酸處理葵百合葉片,系統葉全RNA之北方雜合分析 50 圖四、氨基環丙烷羧酸處理葵百合葉片,處理葉全RNA之北方雜合分析 51 圖五、氨基環丙烷羧酸處理葵百合葉片,系統葉全RNA之北方雜合分析 52 圖六、LsGRP1之5’端快速擴增cDNA末端產物及菌落聚合酵素連鎖反應產物點漬雜合分析 53 圖七、5’端快速擴增cDNA末端法得到LsGRP1相似序列之排並分析 55 圖八、LsGRP1基因體聚合酵素連鎖反應產物之瓊脂精膠體電泳分析及南方雜合分析 56 圖九、LsGRP1基因序列與cDNA序列排並比對 57 圖十、反轉錄聚合酵素連鎖反應產物之瓊脂精膠體電泳分析。(A)LsGRP1;(B)LsGRP1之5’端未轉譯區至終止密碼片段 58 圖十一、LsGRP2與LsGRP1 cDNA序列排並分析 60 圖十二、LsGRP2與LsGRP1之預測胺基酸序列排並分析 61 圖十三、LsGRP1基因體聚合酵素連鎖反應與反轉錄聚合酵素連鎖反應產物之瓊脂精膠體電泳分析 62 圖十四、LsGRP1之3’端快速擴增cDNA末端產物及菌落聚合酵素連鎖反應產物點漬雜合分析 63 圖十五、LsGRP1進行3’端快速增幅cDNA末端得到差異性序列之排並分析 65 圖十六、LsGRP1之3’端快速擴增cDNA末端產物 66 圖十七、LsGRP3與LsGRP1 cDNA序列排並分析 67 圖十八、LsGRP3與LsGRP1之預測胺基酸序列排並分析 68 圖十九、5’、3’端快速增幅cDNA末端選殖株pWSM-2、pWSM-9序列組合與LsGRP1排並分析 70 圖二十、LsGRP4與LsGRP1 cDNA序列排並分析 71 圖二十一、LsGRP4與LsGRP1之預測胺基酸序列排並分析 72 圖二十二、LsGRP4相關序列與LsGRP1基因序列排並分析 75 圖二十三、LsGRP1、LsGRP、LsGRP3、LsGRP4 cDNA序列排並分析 77 拾、附錄 78 圖一、LsGRP1 cDNA核酸序列及其預測胺基酸序列 78 圖二、以LsGRP1探針進行葵百合基因體核酸之南方雜合分析 79 | |
dc.language.iso | zh-TW | |
dc.title | 葵百合LsGRP1及其同源基因之研究 | zh_TW |
dc.title | LsGRP1 and its homologues in Lilium cv. Star Gazer | en |
dc.type | Thesis | |
dc.date.schoolyear | 94-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 鄭石通,賴爾? | |
dc.subject.keyword | 葵百合,glycine-rich-protein,LsGRP1,同源基因, | zh_TW |
dc.subject.keyword | Lilium cv. Star Gazer,glycine-rich protein,LsGRP1,homologues, | en |
dc.relation.page | 79 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2006-01-26 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 植物病理與微生物學研究所 | zh_TW |
顯示於系所單位: | 植物病理與微生物學系 |
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