請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62014
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 王愛玉(Ai-Yu Wang) | |
dc.contributor.author | Fang-Chieh Yang | en |
dc.contributor.author | 楊芳潔 | zh_TW |
dc.date.accessioned | 2021-06-16T13:23:16Z | - |
dc.date.available | 2016-08-29 | |
dc.date.copyright | 2013-08-29 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-07-24 | |
dc.identifier.citation | Bannai H, Tamada Y, Maruyama O, Nakai K, Miyano S (2002) Extensive feature detection of N-terminal protein sorting signals. Bioinformatics 18: 298-305
Bedard J, Jarvis P (2005) Recognition and envelope translocation of chloroplast preproteins. Journal of experimental botany 56: 2287-2320 Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic acids research 7: 1513-1523 Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry 72: 248-254 Chilton MD, Currier TC, Farrand SK, Bendich AJ, Gordon MP, Nester EW (1974) Agrobacterium tumefaciens DNA and PS8 bacteriophage DNA not detected in crown gall tumors. Proceedings of the National Academy of Sciences of the United States of America 71: 3672-3676 Cortese MS, Baird JP, Uversky VN, Dunker AK (2005) Uncovering the unfoldome: enriching cell extracts for unstructured proteins by acid treatment. Journal of proteome research 4: 1610-1618 Dunker AK, Brown CJ, Lawson JD, Iakoucheva LM, Obradovic Z (2002) Intrinsic disorder and protein function. Biochemistry 41: 6573-6582 Dunker AK, Lawson JD, Brown CJ, Williams RM, Romero P, Oh JS, Oldfield CJ, Campen AM, Ratliff CM, Hipps KW, Ausio J, Nissen MS, Reeves R, Kang C, Kissinger CR, Bailey RW, Griswold MD, Chiu W, Garner EC, Obradovic Z (2001) Intrinsically disordered protein. Journal of molecular graphics & modelling 19: 26-59 Emanuelsson O, Nielsen H, Brunak S, von Heijne G (2000) Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. Journal of molecular biology 300: 1005-1016 Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. The Plant journal 6: 271-282 Irar S, Oliveira E, Pages M, Goday A (2006) Towards the identification of late-embryogenic-abundant phosphoproteome in Arabidopsis by 2-DE and MS. Proteomics 6 Suppl 1: S175-185 Jimenez CR, Huang L, Qiu Y, Burlingame AL (2001) In-gel digestion of proteins for MALDI-MS fingerprint mapping. Current protocols in protein science / editorial board, John E. Coligan ... [et al.] Chapter 16: Unit 16 14 Kleffmann T, Russenberger D, von Zychlinski A, Christopher W, Sjolander K, Gruissem W, Baginsky S (2004) The Arabidopsis thaliana chloroplast proteome reveals pathway abundance and novel protein functions. Current biology : CB 14: 354-362 Kleffmann T, von Zychlinski A, Russenberger D, Hirsch-Hoffmann M, Gehrig P, Gruissem W, Baginsky S (2007) Proteome dynamics during plastid differentiation in rice. Plant physiology 143: 912-923 Liao SC, Lin CS, Wang AY, Sung HY (2013) Differential expression of genes encoding Acid invertases in multiple shoots of bamboo in response to various phytohormones and environmental factors. Journal of Agricultural and Food Chemistry 61: 4396-4405 Lopez-Juez E, Pyke KA (2005) Plastids unleashed: their development and their integration in plant development. The International journal of developmental biology 49: 557-577 Oliveira E, Amara I, Bellido D, Odena MA, Dominguez E, Pages M, Goday A (2007) LC-MSMS identification of Arabidopsis thaliana heat-stable seed proteins: enriching for LEA-type proteins by acid treatment. Journal of mass spectrometry : JMS 42: 1485-1495 Radhamony RN, Theg SM (2006) Evidence for an ER to Golgi to chloroplast protein transport pathway. Trends in cell biology 16: 385-387 Rajalingam D, Loftis C, Xu JJ, Kumar TK (2009) Trichloroacetic acid-induced protein precipitation involves the reversible association of a stable partially structured intermediate. Protein science 18: 980-993 Sheen J, Hwang S, Niwa Y, Kobayashi H, Galbraith DW (1995) Green-fluorescent protein as a new vital marker in plant cells. The Plant journal 8: 777-784 Sheerin DJ, Buchanan J, Kirk C, Harvey D, Sun X, Spagnuolo J, Li S, Liu T, Woods VA, Foster T, Jones WT, Rakonjac J (2011) Inter- and intra-molecular interactions of Arabidopsis thaliana DELLA protein RGL1. The Biochemical journal 435: 629-639 Soll J, Schleiff E (2004) Protein import into chloroplasts. Nature reviews. Molecular cell biology 5: 198-208 Sun X, Jones WT, Rikkerink EH (2012) GRAS proteins: the versatile roles of intrinsically disordered proteins in plant signalling. The Biochemical journal 442: 1-12 Tompa P (2010) Structure and function of intrinscially disordered proteins. Chapman & Hall/CRC Press, Boca Raton Tompa P, Csermely P (2004) The role of structural disorder in the function of RNA and protein chaperones. FASEB journal 18: 1169-1175 Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America 76: 4350-4354 Uversky VN (2010) The mysterious unfoldome: structureless, underappreciated, yet vital part of any given proteome. Journal of biomedicine & biotechnology 2010: 568068 Villarejo A, Buren S, Larsson S, Dejardin A, Monne M, Rudhe C, Karlsson J, Jansson S, Lerouge P, Rolland N, von Heijne G, Grebe M, Bako L, Samuelsson G (2005) Evidence for a protein transported through the secretory pathway en route to the higher plant chloroplast. Nature cell biology 7: 1224-1231 Ward JJ, Sodhi JS, McGuffin LJ, Buxton BF, Jones DT (2004) Prediction and functional analysis of native disorder in proteins from the three kingdoms of life. Journal of molecular biology 337: 635-645 Zhang Y, Su J, Duan S, Ao Y, Dai J, Liu J, Wang P, Li Y, Liu B, Feng D, Wang J, Wang H (2011) A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes. Plant methods 7: 30 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62014 | - |
dc.description.abstract | BoMSP41 (Monocot-specific protein-41 in Bambusa oldhamii) 為綠竹之未知功能基因,於綠竹快速生長時期,其基因表現會大幅提升。BoMSP41 胺基酸組成富含 Gln, Glu, Lys, Arg 及 Asp,且含有一些重複的片段。此蛋白質被預測為一個內生性不具穩定構形的蛋白質 intrinsic disorder protein (IDP) 。為了瞭解 BoMSP41 於細胞內存在的位置,本研究建構了以玉米 Ubiquitin 啟動子驅動表現的 GFP::BoMSP41 的重組質體,並對水稻進行轉殖,使 GFP::BoMSP41 融合蛋白質恆常表現於轉殖水稻中。以共軛焦顯微鏡觀察轉殖水稻的原生質體,結果顯示 GFP::BoMSP41 可能被運送至成熟的葉綠體中,推測 BoMSP41 可能參與光合作用、質體基因表現或在葉綠體中發生的其他代謝路徑。為了探討 BoMSP41 的生化性質,以大腸桿菌表現帶有 His-tag 的BoMSP41。由 SDS-聚丙烯醯胺膠體及西方點墨法結果顯示: BoMSP41 於SDS 膠體上的泳動率比預期的分子量應有的泳動率低。此外, BoMSP41 可被 3% TCA沉澱下,但對熱具較高耐受性。泳動率和熱穩定性高這兩點特性與許多 IDP 特性相符。然而,其結構及生化性質仍需進一步探討。 | zh_TW |
dc.description.abstract | BoMSP41 (Monocot-specific protein-41 in Bambusa oldhamii) is an unknown function gene highly up-regulated in rapidly growing bamboo shoots. The BoMSP41-encoding protein contains tandemly repeated sequences and a high proportion of Gln, Glu, Lys, Arg, and Asp. It was predicted to be an intrinsic disorder protein (IDP).
To investigate the localization of BoMSP41 in cells, transgenic rice plants constitutively expressing the GFP::BoMSP41 fusion protein under the control of a maize ubiquitin promoter were constructed. Analysis of protoplasts isolated from seedlings of the transgenic plants by confocal laser scanning microscopy indicated that the GFP::BoMSP41 fusion proteins were transported into mature chloroplasts, suggesting that BoMSP41 may participate in photosynthesis, plastid gene expression or other metabolic pathways occurred in chloroplasts. To characterize the biochemical properties of BoMSP41, the His-tagged BoMSP41 recombinant protein was expressed in Escherichia coli. SDS-PAGE and western analyses showed that the electrophoretic mobility of the recombinant BoMSP41 protein was lower than expected. In addition, the recombinant BoMSP41 protein could be precipitated by 3% TCA but exhibited high thermostability. The lower electrophoretic mobility and high thermostability of BoMSP41 are in agreement with the general properties of many IDPs; however, the structure and the biochemical identity of this protein require further investigation. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T13:23:16Z (GMT). No. of bitstreams: 1 ntu-102-R00b22049-1.pdf: 2216552 bytes, checksum: 0f8776d3dbacc5a8cdfa24a0748e841f (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 目錄 I
縮寫表 III 中文摘要 V Abstract VI 第一章 研究背景 1 1. BoMSP41的緣起 1 2. BoMSP41序列之特殊性 1 3. IDP (Intrinsic disorder protein) 2 3.1 IDP 的特色 2 3.2 IDP 參與之調控與功能 3 4. 本論文之研究目的與主題 4 4.1 GFP::BoMSP41 在水稻轉殖株中之細胞內定位 4 4.2 GFP::BoMSP41 T1 轉殖株於水稻生長速率及表現型的觀察 4 4.3 BoMSP41 蛋白質表現與純化 4 第二章 材料與方法 5 1. 實驗材料 5 1.1 綠竹 5 1.2 轉殖作物 5 1.3 菌種 5 1.4 質體 5 1.5 藥品 6 2. 實驗儀器設備 6 2.1 核酸電泳 6 2.2 蛋白質電泳 6 2.3 離心機 6 2.4 其他儀器 7 3. 實驗方法 7 3.1 BoMSP41 DNA 之選殖 7 3.2 GFP::BoMSP41 融合質體之構築 12 3.3 GFP::BoMSP41-3基因轉殖水稻 13 3.4 T0 代轉殖水稻染色體 DNA 分析 17 3.5 轉殖水稻 T0 代之蛋白質分析 18 3.6 BoMSP41在轉殖水稻內的細胞內定位分析 20 3.7 轉殖水稻 T1 代幼苗型態觀察 21 3.8 pBoMSP41-2 重組蛋白質表現與性質測定 21 3.9 蛋白質身份鑑定 23 第三章 結果 26 1. BoMSP41 基因之選殖與序列分析 26 1.1 BoMSP41 之 基因選殖 26 1.2 BoMSP41-3 胺基酸組成及其特性 26 1.3 BoMSP41-3 之表現產物在細胞內之定位預測 26 2. GFP::BoMSP41 對水稻之轉殖 27 3. GFP::BoMSP41 轉殖株之分析 28 3.1 轉殖株染色體 DNA 分析 28 3.2 轉殖株中 GFP::BoMSP41-3 蛋白質分析 28 3.3 GFP::BoMSP41-3 在轉殖水稻細胞內之定位 28 3.4 GFP::BoMSP41-3 轉殖株 T1 代幼苗型態觀察 29 4. BoMSP41-2 重組蛋白質的表現與性質檢定 29 4.1 重組蛋白質的表現與純化 29 4.2 熱處理與熱穩定性探討 30 4.3 以 TCA 處理測試其酸穩定性 30 第四章 討論 31 1. GFP:: BoMSP41-3 在轉殖水稻細胞內之定位與可能功能 31 2. 重組 BoMSP41-2 蛋白質的表現與性質檢定 32 第五章 結論與未來展望 35 1. 結論 35 2. 未來展望 35 2.1 以不同方法確認 GFP::BoMSP41 之細胞內定位 35 2.2 GFP::BoMSP41 T1植株生長速率之觀察 35 2.3 BoMSP41 抗體的製備和結構分析 36 2.4 與 BoMSP41 交互作用之蛋白質探討 36 2.5 BoMSP41 啟動子的尋找及分析 36 參考文獻 35 圖與表 40 | |
dc.language.iso | zh-TW | |
dc.title | 綠竹 BoMSP41 之細胞內定位與生化性質探討 | zh_TW |
dc.title | Studies on the subcellular localization and biochemical properties of bamboo BoMSP41 | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 洪傳揚(Chwan-Yang Hong) | |
dc.contributor.oralexamcommittee | 宋賢一(Hsien-Yi Sung),王淑珍(Shu-Jen Wang),陳仁治(Jen-Chih Chen),楊健志(Chien-Chih Yang) | |
dc.subject.keyword | 綠竹,單子葉植物特有蛋白質,細胞內定位,葉綠體,內生性不具穩定構形蛋白質, | zh_TW |
dc.subject.keyword | bamboo,monocot-specific protein,subcellular localization,chloroplast,intrinsic disorder protein, | en |
dc.relation.page | 59 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-07-24 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-102-1.pdf 目前未授權公開取用 | 2.16 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。