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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 杜宜殷 | |
dc.contributor.author | Shin-Song You | en |
dc.contributor.author | 尤信淞 | zh_TW |
dc.date.accessioned | 2021-06-16T22:58:53Z | - |
dc.date.available | 2017-08-28 | |
dc.date.copyright | 2012-08-28 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-08-08 | |
dc.identifier.citation | 黃大玲. 2010. 蝴蝶蘭 MYB cDNA 之選殖與分析. 國立臺灣大學生物資源
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/64769 | - |
dc.description.abstract | 為了解蝴蝶蘭轉錄因子 MYB 基因之結構以及其功能,本研究進行蝴
蝶蘭 MYB 基因之選殖。利用蝴蝶蘭轉錄因子 PtMYB1、PtMYB2 與 PtMYB3之 cDNA 作為探針,篩選蝴蝶蘭基因組庫,共獲得 21 個基因組選殖系,其中含有 PtMYB1 之基因組選殖系為 λPtMYB70,而含有 PtMYB2 之基因組選殖系為 λPtMYB261。經次選殖並定序分析後,獲得 PtMYB1 與PtMYB2 之完整基因,皆具有三個顯子與兩個隱子,且隱子邊界序列皆符合GT-AG 規則,此兩基因之解碼區位於第一與第二顯子內的長度皆分別為133 bp 與 130 bp;而啟動子序列經比對可能受到逆境、光照、激勃素、茉莉酸、水楊酸或機械式創傷等因子調控。蛋白質定位的部分,將 PtMYB3 與綠色螢光蛋白 (green fluorescent protien) 構築成融合載體,並於阿拉伯芥原生質體與洋蔥表皮細胞進行暫時性表達。結果顯示均與核染劑 DAPI 完全符合,確認 PtMYB3 蛋白質專一表達於細胞核中。 | zh_TW |
dc.description.abstract | To understand the gene structure and function of the transcription factor MYB from Phalaenopsis, cDNA of PtMYB1, PtMYB2 and PtMYB3 were used as probes, to screen the genomic library of Phalaenopsis. The genomic clone λPtMYB70 and λPtMYB261 contain PtMYB1 gene and PtMYB2 gene, respectively. After subcloning and sequencing, PtMYB1 and PtMYB2 gene both possess three exons and two introns, whose junction sequences all follow the GT-AG rule. The exon 1 and exon 2 are 133 bp and 130 bp, respectively. According to the analysis of the promoter regions of PtMYB1 and PtMYB2, some
cis-acting element related to stress, light, gibberellic acid, methyl jasmonate, salicylic acid, and wounding were found. In order to understand the protein localization, PtMYB3 cDNA fused with green fluorescent protein gene was transiently expressed in Arabidopsis protoplasts and onion epidermal cells. The results of transient assay showed that PtMYB3 specifically expressed in nucleus. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T22:58:53Z (GMT). No. of bitstreams: 1 ntu-101-R99628139-1.pdf: 2968490 bytes, checksum: 7b8c53b2620fd499649275698ddd7c20 (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | 中文摘要 ........................................ I
Abstract ........................................ II 目錄 .............................................III 圖目錄........................................... VI 表目錄 .......................................... VIII 壹、 前言 ....................................... 1 貳、 前人研究 ................................... 2 一、植物轉錄因子 ................................ 2 (一) bZIP 家族 .................................. 2 (二) WRKY 家族 .................................. 3 (三) AP2/EREBP 家族 ............................. 4 (四) MYB 家族 .............................................. 4 二、 植物 MYB 基因之分類與功能 .................. 4 (一) 調節苯基丙酸類生合成路徑 ................... 5 (二) 調節非生物逆境耐受性機制 ................... 10 參、 材料與方法 ................................. 12 一、材料 ........................................ 12 (一) 植物材料 ................................... 12 (二) 基因組庫 ................................... 12 (三) 質體 ....................................... 12 (四) 試驗用之菌種 ............................... 13 二、試驗方法..................................... 13 (一) 基因組庫之篩選與純化 ....................... 13 1. 寄主細胞製備 ................................. 13 2. 核酸探針之製備與標定 ......................... 13 3. 蝴蝶蘭 MYB 基因組庫之選殖..................... 14 4. 噬菌體增殖培養 ............................... 14 5. 噬菌體 DNA 之抽取 ............................ 15 (二) 南方氏雜交分析 ............................. 15 (三) 基因片段之次選殖 ........................... 16 1. DNA 片段回收 ................................. 16 2. 接合反應 (ligation) .......................... 16 3. 勝任細胞 (competent cell) 之製備 .............................................. 17 4. 轉型作用 (transformation) .................... 17 5. 質體小量制備 ................................. 17 (四) 蝴蝶蘭轉錄因子基因啟動子之構築 ............. 18 1. 蝴蝶蘭轉錄因子 PtMYB1 基因啟動子之構築 ....... 18 2. 蝴蝶蘭轉錄因子 PtMYB2 基因啟動子之構築 ....... 18 (五) GFP 融合蛋白之構築 ......................... 19 (六) 原生質體 (protoplast) PEG (polyethylene glycol) 轉殖法 ...... 19 1. 原生質體分離 .............................................. 19 2. PEG 轉殖法 ................................... 20 (七) 基因槍(particle bombardment)轉殖法 ......... 20 1. 金粒子之製備 ................................. 20 2. 基因槍轉殖法 ................................. 21 肆、結果 ........................................ 26 一、 蝴蝶蘭 PtMYB 基因之選殖 .................... 26 (一) 蝴蝶蘭基因組庫之篩選與純化 ................. 26 (二) 蝴蝶蘭 PtMYB1 基因選殖系限制酶圖譜及序列分析..... 26 (三) 蝴蝶蘭轉錄因子 PtMYB1 基因序列分析.......... 33 V (四) 蝴蝶蘭轉錄因子 PtMYB1 .......................33 (五) 蝴蝶蘭轉錄因子 PtMYB1 啟動子暫時性表達分析 ..... 40 (六) 蝴蝶蘭 PtMYB2 基因限制酶圖譜分析 ........... 40 (七) 蝴蝶蘭轉錄因子 PtMYB2 基因序列分析 ......... 40 (八) 蝴蝶蘭轉錄因子 PtMYB2 基因啟動子序列分析 ..... 45 二、蝴蝶蘭 PtMYB3 蛋白質之細胞間定位分析 ........ 51 (一) 阿拉伯芥原生質體之蛋白質定位分析 ........... 51 (二) 洋蔥表皮細胞之蛋白質定位分析 ............... 51 伍、討論 ........................................ 54 一、 蝴蝶蘭轉錄因子 MYB 之基因結構 .............. 54 二、 蝴蝶蘭轉錄因子 MYB 基因之啟動子序列分析 .....54 三、 蝴蝶蘭轉錄因子 MYB 蛋白質定位分析 .......... 59 陸、結語 ........................................ 60 柒、參考文獻 .................................... 61 | |
dc.language.iso | zh-TW | |
dc.title | 蝴蝶蘭 MYB 基因之選殖與蛋白質定位分析 | zh_TW |
dc.title | Cloning and Protein Localization for MYB Genes from
Phalaenopsis | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 黃鵬林 | |
dc.contributor.oralexamcommittee | 黃銓珍,洪傳揚 | |
dc.subject.keyword | 轉錄因子,MYB,基因組選殖系,基因結構,啟動子, | zh_TW |
dc.subject.keyword | transcription factor,MYB,genomic clone,gene structure,promoter, | en |
dc.relation.page | 68 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-08-08 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 園藝學研究所 | zh_TW |
顯示於系所單位: | 園藝暨景觀學系 |
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