請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/24316完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊健志 | |
| dc.contributor.author | Kai-Ting Fan | en |
| dc.contributor.author | 范凱亭 | zh_TW |
| dc.date.accessioned | 2021-06-08T05:21:43Z | - |
| dc.date.copyright | 2005-08-01 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-07-26 | |
| dc.identifier.citation | Andreeva, A.V., and Kutuzov, M.A. (2001). Nuclear localization of the plant protein Ser/Thr phosphatase PP7. Mol Cell Biol Res Commun 4, 345-352.
Barker, S., Laird, S.M., Ho, M.M., Vinson, G.P., and Hinson, J.P. (1992). Characterization of a rat adrenocortical inner zone-specific antigen and identification of its putative precursor. J Mol Endocrinol 9, 95-102. Bauer, J., Hiltbrunner, A., Weibel, P., Vidi, P.A., Alvarez-Huerta, M., Smith, M.D., Schnell, D.J., and Kessler, F. (2002). Essential role of the G-domain in targeting of the protein import receptor atToc159 to the chloroplast outer membrane. J Cell Biol 159, 845-854. Birnboim, H.C., and Doly, J. (1979). A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7, 1513-1523. Chalfie, M., Tu, Y., Euskirchen, G., Ward, W.W., and Prasher, D.C. (1994). Green fluorescent protein as a marker for gene expression. Science 263, 802-805. Czempinski, K., Frachisse, J.M., Maurel, C., Barbier-Brygoo, H., and Mueller-Roeber, B. (2002). Vacuolar membrane localization of the Arabidopsis 'two-pore' K+ channel KCO1. Plant J 29, 809-820. Falkenstein, E., Schmieding, K., Lange, A., Meyer, C., Gerdes, D., Welsch, U., and Wehling, M. (1998). Localization of a putative progesterone membrane binding protein in porcine hepatocytes. Cell Mol Biol (Noisy-le-grand) 44, 571-578. Gerdes, D., Wehling, M., Leube, B., and Falkenstein, E. (1998). Cloning and tissue expression of two putative steroid membrane receptors. Biol Chem 379, 907-911. Gray, W.M., Kepinski, S., Rouse, D., Leyser, O., and Estelle, M. (2001). Auxin regulates SCF(TIR1)-dependent degradation of AUX/IAA proteins. Nature 414, 271-276. Hajdukiewicz, P., Svab, Z., and Maliga, P. (1994). The small, versatile pPZP family of Agrobacterium binary vectors for plant transformation. Plant Mol Biol 25, 989-994. Hand, R.A., and Craven, R.J. (2003). Hpr6.6 protein mediates cell death from oxidative damage in MCF-7 human breast cancer cells. J Cell Biochem 90, 534-547. Hiei, Y., Komari, T., and Kubo, T. (1997). Transformation of rice mediated by Agrobacterium tumefaciens. Plant Mol Biol 35, 205-218. Jefferson, R.A., Burgess, S.M., and Hirsh, D. (1986). beta-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci U S A 83, 8447-8451. Jin, H., Cominelli, E., Bailey, P., Parr, A., Mehrtens, F., Jones, J., Tonelli, C., Weisshaar, B., and Martin, C. (2000). Transcriptional repression by AtMYB4 controls production of UV-protecting sunscreens in Arabidopsis. Embo J 19, 6150-6161. Kao, A.-L., Chang, T.-Y., Chang, S.-H., Su, J.-C., and Yang, C.-C. (2005). Characterization of a novel Arabidopsis protein family AtMAPR homologous to 25-Dx/IZAg/Hpr6.6 proteins. Bot. Bull. Acad. Sin. 46, 107-118. Krebs, C.J., Jarvis, E.D., Chan, J., Lydon, J.P., Ogawa, S., and Pfaff, D.W. (2000). A membrane-associated progesterone-binding protein, 25-Dx, is regulated by progesterone in brain regions involved in female reproductive behaviors. Proc Natl Acad Sci U S A 97, 12816-12821. Liepman, A.H., and Olsen, L.J. (2001). Peroxisomal alanine: glyoxylate aminotransferase (AGT1) is a photorespiratory enzyme with multiple substrates in Arabidopsis thaliana. Plant J 25, 487-498. Mifsud, W., and Bateman, A. (2002). Membrane-bound progesterone receptors contain a cytochrome b5-like ligand-binding domain. Genome Biol 3, RESEARCH0068. Min, L., Takemori, H., Nonaka, Y., Katoh, Y., Doi, J., Horike, N., Osamu, H., Raza, F.S., Vinson, G.P., and Okamoto, M. (2004). Characterization of the adrenal-specific antigen IZA (inner zone antigen) and its role in the steroidogenesis. Mol Cell Endocrinol 215, 143-148. Murashige, T., and Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473-479. Nolte, I., Jeckel, D., Wieland, F.T., and Sohn, K. (2000). Localization and topology of ratp28, a member of a novel family of putative steroid-binding proteins. Biochim Biophys Acta 1543, 123-130. Park, J.H., Oh, S.A., Kim, Y.H., Woo, H.R., and Nam, H.G. (1998). Differential expression of senescence-associated mRNAs during leaf senescence induced by different senescence-inducing factors in Arabidopsis. Plant Mol Biol 37, 445-454. Raza, F.S., Takemori, H., Tojo, H., Okamoto, M., and Vinson, G.P. (2001). Identification of the rat adrenal zona fasciculata/reticularis specific protein, inner zone antigen (IZAg), as the putative membrane progesterone receptor. Eur J Biochem 268, 2141-2147. Rodriguez-Concepcion, M., Toledo-Ortiz, G., Yalovsky, S., Caldelari, D., and Gruissem, W. (2000). Carboxyl-methylation of prenylated calmodulin CaM53 is required for efficient plasma membrane targeting of the protein. Plant J 24, 775-784. Sakamoto, H., Ukena, K., Takemori, H., Okamoto, M., Kawata, M., and Tsutsui, K. (2004). Expression and localization of 25-Dx, a membrane-associated putative progesterone-binding protein, in the developing Purkinje cell. Neuroscience 126, 325-334. Sakamoto, W., Spielewoy, N., Bonnard, G., Murata, M., and Wintz, H. (2000). Mitochondrial localization of AtOXA1, an arabidopsis homologue of yeast Oxa1p involved in the insertion and assembly of protein complexes in mitochondrial inner membrane. Plant Cell Physiol 41, 1157-1163. Selmin, O., Lucier, G.W., Clark, G.C., Tritscher, A.M., Vanden Heuvel, J.P., Gastel, J.A., Walker, N.J., Sutter, T.R., and Bell, D.A. (1996). Isolation and characterization of a novel gene induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in rat liver. Carcinogenesis 17, 2609-2615. Sheen, J., Hwang, S., Niwa, Y., Kobayashi, H., and Galbraith, D.W. (1995). Green-fluorescent protein as a new vital marker in plant cells. Plant J 8, 777-784. Shimomura, O., Johnson, F.H., and Saiga, Y. (1962). Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea. J Cell Comp Physiol 59, 223-239. Takeuchi, M., Ueda, T., Yahara, N., and Nakano, A. (2002). Arf1 GTPase plays roles in the protein traffic between the endoplasmic reticulum and the Golgi apparatus in tobacco and Arabidopsis cultured cells. Plant J 31, 499-515. Walden, R., and Wingender, R. (1995). Gene-transfer and plant-regeneration techniques. Trends in Biotechnology 13, 324-331. Yang, X.H., Xu, Z.H., and Xue, H.W. (2005). Arabidopsis membrane steroid binding protein 1 is involved in inhibition of cell elongation. Plant Cell 17, 116-131. Yu, T.S., and Li, H. (2001). Chloroplast protein translocon components atToc159 and atToc33 are not essential for chloroplast biogenesis in guard cells and root cells. Plant Physiol 127, 90-96. Zeng, G. (1998). Sticky-end PCR: new method for subcloning. Biotechniques 25, 206-208. Zimmer, M. (2002). Green fluorescent protein (GFP): applications, structure, and related photophysical behavior. Chem Rev 102, 759-781. 張碩修. (2003). 阿拉伯芥中與 AtMAPR有交互作用的蛋白質之篩選 (國立台灣大學農業化學研究所碩士論文). 高艾玲. (2002). 阿拉伯芥中 MAPR 同源蛋白質基因之選殖、表現與功能分析 (國立台灣大學農業化學研究所碩士論文). 劉耿全. (2005). 以酵母菌雙雜合系統篩選阿拉伯芥中與AtMAPRs有交互作用之蛋白質 (國立台灣大學微生物與生化學研究所碩士論文) 蔡佩璇. (2004). 阿拉伯芥中 AtMAPRs 基因靜默載體建構與表現分析 (國立台灣大學微生物與生化學研究所碩士論文). | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/24316 | - |
| dc.description.abstract | 我們在阿拉伯芥中發現四個豬的MAPR (putative membrane-associated progesterone receptor component 1) 同源蛋白質，分別為AtMAPR2、AtMAPR3、AtMAPR4 以及AtMAPR5。在先前的研究中指出，位於細胞膜上之AtMAPR5 (putative membrane steroid binding protein, MSBP1) 可在胞外與黃體素及芸苔素內酯結合，且黑暗培養可抑制在下胚軸中該基因之表現，受到光照則可回復表現，推測在阿拉伯芥中功能為細胞延長之負調控因子。AtMAPRs家族蛋白質中其他三者的功能則尚未明瞭，因此嘗試以細胞、組織定位及觀察過量表現AtMAPRs基因轉殖株之性狀，探討AtMAPRs可能之生理意義。在本論文中利用基因槍將AtMAPR2與GFP之融合基因轉殖於洋蔥表皮細胞，由GFP螢光位置以得到AtMAPR2在細胞中之定位，結果發現AtMAPR2雖然有較為聚集在細胞核內之情形，但也散佈於細胞質中；而使用GUS報導基因的暫時性表現結果也間接說明AtMAPR2並非核蛋白。另一方面，利用農桿菌轉殖法得到過量表現AtMAPR2-GUS-GFP之阿拉伯芥T2轉殖株，使用共軛焦顯微鏡觀察其幼苗之根部及下胚軸的螢光表現，發現AtMAPR2會聚集在細胞中的某種胞器內且並非細胞核。而藉由GUS染色發現，經過不同光照處理培養之T2轉殖株中，AtMAPR2在下胚軸處也許可藉由全日光照而增加其蛋白質之穩定性，且可能在植株發育的前四天即決定AtMAPR2之穩定性，之後便無法以光照或黑暗對其穩定與否產生影響。至於藉由不同賀爾蒙處理下，過量表現AtMAPR2轉殖株根部的發育，推測其直接或間接參與了由auxin所調控之根部延長。這些結果不但有助於釐清AtMAPRs家族之可能生理意義，未來更可將過量表現AtMAPR2之轉殖株與RNAi或T-DNA突變株進行雜交，以期能藉由觀測性狀改變而確定其可能之參與之訊息傳遞路徑。 | zh_TW |
| dc.description.abstract | Four porcine MAPR (putative membrane-associated progesterone receptor component 1) homologs in Arabidopsis were named as AtMAPR2, AtMAPR3, AtMAPR4, and AtMAPR5. In recent study, it is demonstrated that AtMAPR5 (also defined as putative membrane steroid binding protein, MSBP1) localizes to plasma membrane and binds to progesterone and brassinolide in vivo. The expression of AtMAPR5 in hypocotyls is inhibited in the dark, and is activated under continuous illumination. It suggests that AtMAPR5 is a negative regulator of cell elongation. However, the functions of other homologs are still unclear. In this study, we tried to find the possible functions of AtMAPRs by the tissue or subcellular distribution of GFP or GUS-fusion protein expressed in transgenic plants followed by phenotype analysis. Using particle bombardment, the transient expression of AtMAPR2-GFP fusion protein in onion epidermal cells showed that although in some cases AtMAPR2 aggregated in the nucleus, the cytoplasm also contained the green fluorescence. Besides, the transient expression of AtMAPR2-GUS in onion cells suggested that AtMAPR2 is not a kind of nuclear protein. We established a transgenic T2 plant overexpressing AtMAPR2-GUS-GFP by Agrobacterium-mediated transformation. It is interesting that in the roots and hypocotyls of transgenic seedlings the AtMAPR2-GUS-GFP was located in some organelles but may not be in nucleus as seen under confocal laser microscope. The GUS staining analysis of transgenic T2 seedlings grown under different light conditions demonstrated that light might increase the stability of AtMAPR2 in hypocotyls during the first four days following germination. And the stability would not be abolished or complemented by the darkness or light for the 4-d-old seedlings. The possible function of AtMAPR2 could be predicted by observing the roots of transgenic plants grown on vertical medium containing different hormones. AtMAPR2 might be involved in the root elongation regulated by auxin. These results can help us to understand the possible roles of AtMAPRs in the signal transduction. The overexpressing-AtMAPR2 transgenic T2 plants can be used to cross with RNAi or T-DNA mutants in the future and help us to further identify its function. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T05:21:43Z (GMT). No. of bitstreams: 1 ntu-94-R92b47205-1.pdf: 6722471 bytes, checksum: 09f6c5be33ba7df02ed3dc0083680b28 (MD5) Previous issue date: 2005 | en |
| dc.description.tableofcontents | 目錄………………………………………...………………………………………….I
縮寫表………………………………...……………………………………………..IV 摘要………………………………………………………………………………...VI Abstract………………………………………………………………………..……VII 第一章 緒論…………………………………………………………………………1 1.1 AtMAPRs家族蛋白質…………………………………………………......1 1.2 MAPR家族蛋白質在細胞定位上之研究………………………………....2 1.3 組織及細胞定位中常用之報導基因……………………………………...2 1.3.1 β-glucuronidase (GUS)……………………………………………….3 1.3.2 綠色螢光蛋白 (Green fluorescent protein)……………………….…4 1.4 基因轉形 (Transformation)………………………………………………..6 1.4.1 農桿菌 (Agrobacterium tumefaciens) 轉殖機制………………...….6 1.4.2 基因槍基因轉殖 (Particle bombardment)..........................................9 1.5 Binary vector介紹……………………………………………………….10 1.6 研究動機、目的與方向…………………………………………………...11 第二章 材料與方法………………………………………………………………..12 2.1 實驗材料………………………………………………………………….12 2.1.1 植物材料……………………………………………………………12 2.1.2 載體 (vectors)………………………………………………………12 2.1.3 大腸桿菌菌株………………………………………………………14 2.1.4 農桿菌菌株…………………………………………………………14 2.2 實驗藥品………………………………………………………………….15 2.2.1 一般化學藥劑………………………………………………………15 2.2.2 酵素…………………………………………………………………15 2.2.3 培養基………………………………………………………………15 2.3 儀器設備……………………………………………………………….....16 2.4 實驗方法………………………………………………………………….17 2.4.1 阿拉伯芥種植………………………………………………………17 2.4.1.1 培養基之配製………………………………………………...17 2.4.1.2 種子之表面消毒與低溫處理………………………………...19 2.4.1.3 種子之無菌培養……………………………………………...19 2.4.1.4 土壤培養……………………………………………………...20 2.4.1.5 收集種子……………………………………………………...20 2.4.2 DNA之抽取與分析………………………………………………...20 2.4.2.1 阿拉伯芥染色體DNA之抽取……………………………….20 2.4.2.2 質體DNA之抽取與分析…………………………………….21 2.4.2.3 DNA限制酶切割分析方法…………………………………..23 2.4.2.4 洋菜膠體電泳………………………………………………...23 2.4.2.5 DNA片段的分離及純化……………………………………..24 2.4.2.6 DNA之去磷酸化反應………………………………………..24 2.4.2.7 DNA之磷酸化反應…………………………………………24 2.4.2.8 DNA的定量…………………………………………………..24 2.4.3 AtMAPR基因序列選殖……………………………………………..25 2.4.3.1 專一性引子設計 .....................................................................25 2.4.3.2 聚合酶連鎖反應 (Polymerase chain reaction, PCR) ………26 2.4.3.3 T-A cloning………………………………………………...….26 2.4.3.4 大腸桿菌之質體轉形………………………………………...27 2.4.4 表現載體之建構…………………………………………………28 2.4.4.1 Binary vector之建構………………………………………….28 2.4.4.2 暫時性基因表現 (transient expression) 載體之建…………29 2.4.5 阿拉伯芥基因轉殖………………………………………………31 2.4.5.1 植物材料之準備……………………………………………...31 2.4.5.2 農桿菌轉形…………………………………………………...32 2.4.5.3 農桿菌感染—花序浸潤法 (floral dip method)………….......33 2.4.5.4 轉殖株之篩選………………………………………………...34 2.4.6 基因槍 (bombardment) 轉殖………………………………………35 2.4.6.1 植物材料之準備……………………………………………...35 2.4.6.2 DNA包裹鎢粒子之處理…………………………………......36 2.4.6.3 操作基因槍…………………………………………………...36 2.4.6.4 螢光染劑propidium iodide (PI) 之染色……………………37 2.4.7 觀察T2轉殖株與暫時性表現之洋蔥表皮細胞…………………...38 2.4.7.1 不同處理下組織之GUS染色………………………………38 2.4.7.2 垂直培養T2植株之根系發育觀察…………………………38 2.4.7.3 顯微鏡及影像分析……………………………………….......39 第三章 結果與討論……………………………………………………………......41 3.1 AtMAPRs基因序列選殖………………………………………………….41 3.2 表現載體建構…………………………………………………………….41 3.3 阿拉伯芥之基因轉殖…………………………………………………….43 3.3.1 第一代轉殖株 (T1) 之篩選與培養…………………………….....44 3.3.2 第二代轉殖株 (T2) 之培養與GUS染色分析…………………....46 3.4 光照及其他處理對AtMAPR2的影響…………………………………...47 3.5 過度表現AtMAPR2對阿拉伯芥根系生長發育之影響………………...50 3.6 AtMAPR2在細胞中的定位………………………………………………51 3.6.1 洋蔥表皮細胞之暫時表現…………………………………………51 3.6.2 CaMV35S::AtMAPR2-GUS-mGF5P之T2轉殖株……………….52 第四章 結論與未來展望…………………………………………………………..53 參考文獻………………………………………………………………..……………55 圖與表………………………………………………………………………………..59 附錄………………………………………………………………………………......83 口試問答摘要………………………………………………………………..………88 | |
| dc.language.iso | zh-TW | |
| dc.subject | 定位研究 | zh_TW |
| dc.subject | 芸苔素內酯 | zh_TW |
| dc.subject | 阿拉伯芥 | zh_TW |
| dc.subject | 黃體素 | zh_TW |
| dc.subject | 固醇類賀爾蒙受體 | zh_TW |
| dc.subject | mPR | en |
| dc.subject | Arabidopsis | en |
| dc.subject | brassinosteroid | en |
| dc.subject | localization | en |
| dc.subject | AtMAPRs | en |
| dc.subject | MAPR | en |
| dc.subject | progesterone | en |
| dc.title | 阿拉伯芥AtMAPRs之組織及細胞內定位探討 | zh_TW |
| dc.title | Tissue and Subcellular Localization of AtMAPRs from Arabidopsis | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蘇仲卿,靳宗洛,吳素幸,陳佩燁 | |
| dc.subject.keyword | 固醇類賀爾蒙受體,黃體素,芸苔素內酯,阿拉伯芥,定位研究, | zh_TW |
| dc.subject.keyword | AtMAPRs,MAPR,mPR,progesterone,brassinosteroid,Arabidopsis,localization, | en |
| dc.relation.page | 94 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2005-07-26 | |
| 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 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-94-1.pdf 未授權公開取用 | 6.56 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。