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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 李昆達(Kung-Ta Lee) | |
dc.contributor.author | Han-Jung Kuo | en |
dc.contributor.author | 郭翰蓉 | zh_TW |
dc.date.accessioned | 2021-06-15T04:57:38Z | - |
dc.date.available | 2011-08-23 | |
dc.date.copyright | 2011-08-23 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-08-17 | |
dc.identifier.citation | Burlat V, Kwon M, Davin LB, Lewis NG (2001) Dirigent proteins and dirigent sites in lignifying tissues. Phytochemistry 57: 883-897
Chiu W, Niwa Y, Zeng W, Hirano T, Kobayashi H, Sheen J (1996) Engineered GFP as a vital reporter in plants. Curr Biol 6: 325-330 Chung CT, Niemela SL, Miller RH (1989) One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci U S A 86: 2172-2175 Davin LB, Lewis NG (2000) Dirigent proteins and dirigent sites explain the mystery of specificity of radical precursor coupling in lignan and lignin biosynthesis. Plant Physiol 123: 453-462 Davin LB, Wang HB, Crowell AL, Bedgar DL, Martin DM, Sarkanen S, Lewis NG (1997) Stereoselective bimolecular phenoxy radical coupling by an auxiliary (dirigent) protein without an active center. Science 275: 362-366 Dewick AJBaPM (1990a) Aryltetralin lignans from Linum flavum and Linum capitatum. Phytochemistry 29: 3839-3844 Dewick AJBaPM (1990b) Tumour-inhibitory aryltetralin lignans in Podophyllum versipelle, Diphylleia cymosa and Diphylleia grayi. Phytochemistry 29: 3831-3837 Dinkova-Kostova AT, Gang DR, Davin LB, Bedgar DL, Chu A, Lewis NG (1996) (+)-Pinoresinol/(+)-lariciresinol reductase from Forsythia intermedia. Protein purification, cDNA cloning, heterologous expression and comparison to isoflavone reductase. J Biol Chem 271: 29473-29482 Drummond AJ AB, Buxton S, Cheung M, Cooper A, Duran C, Field M, Heled J, Kearse M, Markowitz S, Moir R, Stones-Havas S, Sturrock S, Thierer T, Wilson A (2010) Geneious v5.3. Available from http://www.geneious.com Farkya S, Bisaria VS, Srivastava AK (2004) Biotechnological aspects of the production of the anticancer drug podophyllotoxin. Appl Microbiol Biotechnol 65: 504-519 Fujita M, Gang DR, Davin LB, Lewis NG (1999) Recombinant pinoresinol-lariciresinol reductases from western red cedar (Thuja plicata) catalyze opposite enantiospecific conversions. J Biol Chem 274: 618-627 Gang DR, Costa MA, Fujita M, Dinkova-Kostova AT, Wang HB, Burlat V, Martin W, Sarkanen S, Davin LB, Lewis NG (1999) Regiochemical control of monolignol radical coupling: a new paradigm for lignin and lignan biosynthesis. Chem Biol 6: 143-151 Halls SC, Davin LB, Kramer DM, Lewis NG (2004) Kinetic study of coniferyl alcohol radical binding to the (+)-pinoresinol forming dirigent protein. Biochemistry 43: 2587-2595 Halls SC, Lewis NG (2002) Secondary and quaternary structures of the (+)-pinoresinol-forming dirigent protein. Biochemistry 41: 9455-9461 Hande KR (1998) Etoposide: four decades of development of a topoisomerase II inhibitor. Eur J Cancer 34: 1514-1521 Hemmati S, Schmidt TJ, Fuss E (2007) (+)-Pinoresinol/(-)-lariciresinol reductase from Linum perenne Himmelszelt involved in the biosynthesis of justicidin B. FEBS Lett 581: 603-610 Kim MK, Jeon JH, Fujita M, Davin LB, Lewis NG (2002) The western red cedar (Thuja plicata) 8-8' DIRIGENT family displays diverse expression patterns and conserved monolignol coupling specificity. Plant Mol Biol 49: 199-214 Konuklugil B (1996) Investigation of podophyllotoxin in some plants in Lamiaceae using HPLC. J Fac Pharm Ankara 25: 23-27 Kupchan SM, Hemingway JC, Knox JR (1965) Tumor inhibitors. VII. Podophyllotoxin, the active principle of Juniperus virginiana. J Pharm Sci 54: 659-660 Min T, Kasahara H, Bedgar DL, Youn B, Lawrence PK, Gang DR, Halls SC, Park H, Hilsenbeck JL, Davin LB, Lewis NG, Kang C (2003) Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases. J Biol Chem 278: 50714-50723 Nakatsubo T, Mizutani M, Suzuki S, Hattori T, Umezawa T (2008) Characterization of Arabidopsis thaliana pinoresinol reductase, a new type of enzyme involved in lignan biosynthesis. J Biol Chem 283: 15550-15557 Novelo M, Cruz JG, Hernandez L, Pereda-Miranda R, Chai H, Mar W, Pezzuto JM (1993) Cytotoxic constituents from Hyptis verticillata. J Nat Prod 56: 1728-1736 von Heimendahl CB, Schafer KM, Eklund P, Sjoholm R, Schmidt TJ, Fuss E (2005) Pinoresinol-lariciresinol reductases with different stereospecificity from Linum album and Linum usitatissimum. Phytochemistry 66: 1254-1263 Walker A, Taylor J, Rowe D, Summers D (2008) A method for generating sticky-end PCR products which facilitates unidirectional cloning and the one-step assembly of complex DNA constructs. Plasmid 59: 155-162 Xia ZQ, Costa MA, Pelissier HC, Davin LB, Lewis NG (2001) Secoisolariciresinol dehydrogenase purification, cloning, and functional expression. Implications for human health protection. J Biol Chem 276: 12614-12623 Xia ZQ, Costa MA, Proctor J, Davin LB, Lewis NG (2000) Dirigent-mediated podophyllotoxin biosynthesis in Linum flavum and Podophyllum peltatum. Phytochemistry 55: 537-549 盧佩君 (2008) 台灣八角蓮 Pinoresinol lariciresinol reductase 與 Secoisolariciresinol dehydrogenase 之基因選殖與表現. Gene Cloning and Heterologous Expression of Pinoresinol Lariciresinol Reductase and Secoisolariciresinol Dehydrogenase Gene from Podophyllum pleianthum Hance. 臺灣大學 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46197 | - |
dc.description.abstract | 鬼臼素生合成路徑之起始步驟為兩個分子的松柏醇自由基經由方向蛋白 (dirigent protein, DP) 的作用,進行方向性之雙聚體化 (dimerization),以形成一分子之松脂醇。自臺灣八角蓮之RNA經反轉錄選殖出一579鹼基對長之方向蛋白基因,其轉譯出之蛋白質預測分子量為22 kDa。該基因在核酸序列上與美洲八角蓮之方向蛋白具97%之一致性,其胺基酸序列則分別有96%之一致性及98%之相似度。此方向蛋白以大腸桿菌進行異源表現時無法產生功能性蛋白質,而在菸草 (Nicotiana tabacum) 毛狀根之植物異源表現系統中,以螢光顯微鏡可觀察到DP-ZsYellow之螢光表現。以粗蛋白進行松柏醇雙聚體化反應,則可觀測到產物松脂醇之生成,且被進一步轉換為下游代謝物。 | zh_TW |
dc.description.abstract | Podophyllotoxin is an important precursor of anti-tumor drug. Podophylloum pleianthum Hance, an endemic species in Taiwan, is a nature source of podophyllotoxin. The biosynthesis pathway of podophyllotoxin starts from two molecules of coniferyl alcohol radical coupled directionally forming one molecule of (+)-pinoresinol with the involvement of dirigent protein (DP). The cDNA of dirigent protein was cloned from P. pleianthum Hance total RNA by RT-PCR. The length of the dirigent protein cDNA was 579 base pairs and the predicted molecular weight of the dirigent protein was 22 kDa. The cDNA sequence was aligned with DP of Podophyllum peltatum. The identity was 97%. The identity and similarity of amino acid alignment were 96% and 98% respectively. The dirigent protein could not express functionally in recombinant E. coli system. Nicotiana tabacum hairy root for heterologous expression was taken instead. The fluorescent reporter protein ZsYellow was observed by fluorescent microscope. The dimerization reaction of coniferyl alcohol in vitro was performed. Pinoresinol was first converted and was then transferred to other metabolites. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T04:57:38Z (GMT). No. of bitstreams: 1 ntu-100-R97B47103-1.pdf: 2527978 bytes, checksum: 92465875e54b27e0160b1f63362b3c4c (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 口試委員審定書 i
致謝 ii 摘要 iii Abstract iv Abbreviation v Contents vi Contents of figures viii Contents of tables ix 1. Introduction 1 2. Materials and methods 6 2.1 Organisms 6 2.2 E. coli plasmid DNA extraction 6 2.3 Plant genomic DNA extraction 7 2.4 Plant RNA extraction 8 2.5 dp cloning 8 2.6 Sequence alignment 9 2.7 E. coli expression 9 2.8 E. coli protein extraction and purification 10 2.9 ps35ScYellow construction 11 2.10 Expression vector po35S-dp construction 12 2.11 Expression vector ps35S-dpcYellw construction 12 2.12 Agrobacterium transformation 13 2.13 A. rhizogenes A4 infection and hairy root transformation 14 2.14 Protein extraction of plant 15 2.15 Western blot analysis 15 2.16 Dirigent protein enzyme assay 16 2.17 High performance liquid chromatography (HPLC) analysis 16 3. Results 18 3.1 dp cloning 18 3.2 E. coli expression 18 3.3 N. tabacum hairy root expression 19 4. Discussion 21 5. Perspective 24 Figures 25 Tables 42 References 43 Appendix 47 | |
dc.language.iso | en | |
dc.title | 臺灣八角蓮方向蛋白基因之選殖與表現 | zh_TW |
dc.title | Cloning and Expression of Dirigent Protein from Podophyllum pleianthum Hance | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 蘇遠志,黃鵬林,賴宗賢 | |
dc.subject.keyword | 鬼臼素,臺灣八角蓮,方向蛋白,松柏醇,松脂醇, | zh_TW |
dc.subject.keyword | Podophyllotoxin,Podophyllum pleianthum Hance,Dirigent protein,Coniferyl alcohol,Pinoresinol, | en |
dc.relation.page | 50 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-08-18 | |
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 |
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