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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 | Yu Huang | en |
dc.contributor.author | 黃鈺 | zh_TW |
dc.date.accessioned | 2021-06-15T00:20:17Z | - |
dc.date.available | 2011-08-18 | |
dc.date.copyright | 2011-08-18 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-08-15 | |
dc.identifier.citation | An G, Watson BD, Chiang CC (1986) Transformation of Tobacco, Tomato, Potato, and Arabidopsis thaliana Using a Binary Ti Vector System. Plant Physiol 81:301-305
Andersen DC, Krummen L (2002) Recombinant protein expression for therapeutic applications. Curr Opin Biotechnol 13:117-123 Binns AN (2002) T-DNA of Agrobacterium tumefaciens: 25 years and counting. Trends Plant Sci 7:231 Boevink P, Martin B, Oparka K, Santa Cruz S, Hawes C (1999) Transport of virally expressed green fluorescent protein through the secretory pathway in tobacco leaves is inhibited by cold shock and brefeldin A. Planta 208:392-400 Bross P, Andersen BA, Winter V, Krautle F, Jensen TG, Nandy A, Kolvraa S, Ghisla S, Bolund L, Gregersen N (1993) Co-overexpression of bacterial GroESL chaperonins partly overcomes non-productive folding and tetramer assembly of E. coli-expressed human medium-chain acyl-CoA dehydrogenase (MCAD) carrying the prevalent disease-causing K304E mutation. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease 1182:264-274 Capell T, Christou P (2004) Progress in plant metabolic engineering. Curr Opin Biotechnol 15:148-154 Chen S, Li X, Liu X, Xu H, Meng K, Xiao G, Wei X, Wang F, Zhu Z (2005) Green fluorescent protein as a vital elimination marker to easily screen marker-free transgenic progeny derived from plants co-transformed with a double T-DNA binary vector system. Plant Cell Rep 23:625-631 Chen SC, Liu HW, Lee KT, Yamakawa T (2007) High-efficiency Agrobacterium rhizogenes-mediated transformation of heat inducible sHSP18. 2-GUS in Nicotiana tabacum. Plant Cell Rep 26:29-37 Chiang PY (2006) Expression of green fluorescent protein in tobacco hairy roots. master thesis, National Taiwan University Cho HJ, Farrand SK, Noel GR, Widholm JM (2000) High-efficiency induction of soybean hairy roots and propagation of the soybean cyst nematode. Planta 210:195-204 Choi H, Son J, Na G, Hong S, Park Y, Song J (2002) Mass production of paclitaxel by plant cell culture. Korean J Plant Biotechnol 29:59!V62 Collier R, Fuchs B, Walter N, Kevin Lutke W, Taylor CG (2005) Ex vitro composite plants: an inexpensive, rapid method for root biology. The Plant Journal 43:449-457 Cubitt AB, Heim R, Adams SR, Boyd AE, Gross LA, Tsien RY (1995) Understanding, improving and using green fluorescent proteins. Trends Biochem Sci 20:448-455 Daley M, Knauf V, Summerfelt K, Turner J (1998) Co-transformation with one Agrobacterium tumefaciens strain containing two binary plasmids as a method for producing marker-free transgenic plants. Plant Cell Rep 17:489-496 De Cleene M, De Ley J (1976) The host range of crown gall. The Botanical Review 42:389-466 DiCosmo F, Misawa M (1985) Eliciting secondary metabolism in plant cell cultures. Trends Biotechnol 3:318-322 DiCosmo F, Misawa M (1995) Plant cell and tissue culture: alternatives for metabolite production. Biotechnol Adv 13:425-453 Doran PM (2000) Foreign protein production in plant tissue cultures. Curr Opin Biotechnol 11:199-204 Finnegan J, McElroy D (1994) Transgene inactivation: plants fight back! Nat Biotechnol 12:883-888 Ganapathi B, Kargi F (1990) Recent advances in indole alkaloid production by Catharanthus roseus (Periwinkle). J Exp Bot 41:259 Gelvin SB (1998) The introduction and expression of transgenes in plants. Curr Opin Biotechnol 9:227-232 Guillon S, Tremouillaux-Guiller J, Pati PK, Rideau M, Gantet P (2006) Hairy root research: recent scenario and exciting prospects. Curr Opin Plant Biol 9:341-346 Heim R, Tsien RY (1996) Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer. Curr Biol 6:178-182 Hellwig S, Drossard J, Twyman RM, Fischer R (2004) Plant cell cultures for the production of recombinant proteins. Nat Biotechnol 22:1415-1422 Hezari M, Croteau R (1997) Taxol biosynthesis: an update. Planta Med 63:291-295 James E, Lee J (2001) The production of foreign proteins from genetically modified plant cells. Plant Cells 127-156 Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Report 5:387-405 Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. The EMBO journal 6:3901 Komari T, Hiei Y, Saito Y, Murai N, Kumashiro T (1996) Vectors carrying two separate T-DNAs for co-transformation of higher plants mediated by Agrobacterium tumefaciens and segregation of transformants free from selection markers. Plant J 10:165-174 Komari T, Takakura Y, Ueki J, Kato N, Ishida Y, Hiei Y (2006) Binary vectors and super-binary vectors. Methods Mol Biol 343:15-41 Lee JM, Hatzimanikatis V (1992) Biochemical engineering. Prentice Hall Li C, Schwabe JWR, Banayo E, Evans RM (1997) Coexpression of nuclear receptor partners increases their solubility and biological activities. Proc Natl Acad Sci U S A 94:2278 Liu HW (2004) Expression of a heat inducible promoter of Arabidopsis in tobacco hairy roots. master thesis, National Taiwan University Matthews PR, Wang MB, Waterhouse PM, Thornton S, Fieg SJ, Gubler F, Jacobsen JV (2001) Marker gene elimination from transgenic barley, using co-transformation with adjacenttwin T-DNAs' on a standard Agrobacterium transformation vector. Mol Breed 7:195-202 McCormac AC, Fowler MR, Chen DF, Elliott MC (2001) Efficient co-transformation of Nicotiana tabacum by two independent T-DNAs, the effect of T-DNA size and implications for genetic separation. Transgenic Res 10:143-155 McCoy E, O'Connor SE (2008) Natural products from plant cell cultures. Prog Drug Res 65:329-370 Miller M, Tagliani L, Wang N, Berka B, Bidney D, Zhao ZY (2002) High efficiency transgene segregation in co-transformed maize plants using an Agrobacterium tumefaciens 2 T-DNA binary system. Transgenic Res 11:381-396 Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473-497 Novick RP (1987) Plasmid incompatibility. Microbiol Mol Biol Rev 51:381 Ohta S, Mita S, Hattori T, Nakamura K (1990) Construction and expression in tobacco of a b-glucuronidase (GUS) reporter gene containing an intron within the coding sequence. Plant Cell Physiol 31:805 Palazon J, Mallol A, Eibl R, Lettenbauer C, Cusido RM, Pinol MT (2003) Growth and ginsenoside production in hairy root cultures of Panax ginseng using a novel bioreactor. Planta Med 69:344-349 Parr A, Peerless A, Hamill J, Walton N, Robins R, Rhodes M (1988) Alkaloid production by transformed root cultures of Catharanthus roseus. Plant Cell Rep 7:309-312 Rivas FV, Tolia NH, Song JJ, Aragon JP, Liu J, Hannon GJ, Joshua-Tor L (2005) Purified Argonaute2 and an siRNA form recombinant human RISC. Nat Struct Mol Biol 12:340-349 Shanks JV, Morgan J (1999) Plant 'hairy root'culture. Curr Opin Biotechnol 10:151-155 Sripriya R, Raghupathy V, Veluthambi K (2008) Generation of selectable marker-free sheath blight resistant transgenic rice plants by efficient co-transformation of a cointegrate vector T-DNA and a binary vector T-DNA in one Agrobacterium tumefaciens strain. Plant Cell Rep 27:1635-1644 Su CY (2009) Agrobacterium rhizogenes-mediated transformation of Nicotiana tabacum Wisconsin 38. master thesis, National Taiwan University Sun L, Zhou L, Lu M, Cai M, Jiang XW, Zhang QX (2009) Marker-Free Transgenic Chrysanthemum Obtained by Agrobacterium-Mediated Transformation with Twin T-DNA Binary Vectors. Plant Mol Biol Report 27:102-108 Taketomi E, Silva D, Sopelete M, Gervasio A, Alves R, Sung S (2006) Differential IgE reactivity to Der p 1 and Der p 2 allergens of. J Investig Allergol Clin Immunol 16:104-109 Tyo KEJ, Ajikumar PK, Stephanopoulos G (2009) Stabilized gene duplication enables long-term selection-free heterologous pathway expression. Nat Biotechnol 27:760-765 Tzfira T, Citovsky V (2006) Agrobacterium-mediated genetic transformation of plants: biology and biotechnology. Curr Opin Biotechnol 17:147-154 Verpoorte R, Contin A, Memelink J (2002) Biotechnology for the production of plant secondary metabolites. Phytochem Rev 1:13-25 Wani MC, Taylor HL, Wall ME, Coggon P, McPhail AT (1971) Plant antitumor agents. VI. Isolation and structure of taxol, a novel antileukemic and antitumor agent from Taxus brevifolia. J Am Chem Soc 93:2325-2327 Yamamura Y, Sahin FP, Nagatsu A, Mizukami H (2003) Molecular cloning and characterization of a cDNA encoding a novel apoplastic protein preferentially expressed in a shikonin-producing callus strain of Lithospermum erythrorhizon. Plant Cell Physiol 44:437 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/41476 | - |
dc.description.abstract | 為了建立以根毛農桿菌介導轉形多基因的方法,我們將報導基因各建構於不同的雙元質體,分別由兩菌體攜帶(2AR)或由單一菌體攜帶(2BV),另外也設計於同一雙元質體中將兩不同報導基因置於同一T-DNA中(1TD)或是不同的T-DNA中(2TD)對菸草進行感染與轉形研究。比較此四種方法之誘導率、DNA轉形效率、以及異源蛋白GUS與GFP的表現量,結果顯示,四種方法的毛狀根誘導率皆達到九成以上,且無異於野生型根毛農桿菌 (Agrobacterium rhizogenes) 的誘導率。2AR、2BV、1TD、2TD的轉形效率分別為65.4%、40%、78.6%、82.1%,以1TD與2TD的轉形效率較佳。另外,結果顯示此四種策略均可在菸草毛狀根中共表現GUS與GFP,。在轉形毛狀根的表現方面, 2AR、2BV、1TD、2TD的GUS活性分別介於0~259.1、0.2~698.9、0.7~112.3、0.6~698.9 nmole MU min-1 mg protein-1;在GFP表現量方面,則分別介於5.5~6333.9、0.4~8758.7、1.3~1923.5、0.7~8459.3 ng mg-1 protein。綜合來看,1TD與2TD是較好的共轉形及共表現組,但四種轉形策略皆可對植物進行多基因的轉形。 | zh_TW |
dc.description.abstract | To establish a platform of Agrobacterium rhizogenes-mediated multiple-gene transformation, we proposed and evaluated different strategies in this study. Two reporter genes are carried by two separate plasmids within different transformed A. rhizogenes (2AR) and within one transformed A. rhizogenes (2BV). Two reporter genes are also constructed in one T-DNA (1TD) and in different T-DNA (2TD) in one binary vector. In the study, we compare the hairy root inducing rate, genes co-transforming rate and the expression of reporter genes-GUS and GFP. Hairy root induction rate of the four groups were all over 90%, without difference from wild type A. rhizogenes. The transformation efficiency of 2AR, 2BV, 1TD and 2TD was 65.4%, 40%, 78.6%, and 82.1%, respectively. Our results showed that all of the four strategies can co-express GUS and GFP in tobacco hairy roots. The GUS expression of 2AR, 2BV, 1TD and 2TD ranged from 0 to 259.1, from 0.2 to 698.9, from 0.7 to 112.3, and from 0.6 to 698.1 nmol MU min-1 mg-1 protein, respectively. Meanwhile, the transgenic hairy roots expressed GFP ranging from 5.5 to 6333.9, from 0.4 to 8758.7, from 1.3 to 1923.5, and from 0.7 to 8459.3 ng mg-1 protein, respectively. 1TD and 2TD were better strategies to gain co-expression hairy roots, while all of the four strategies were able to transform multiple genes into plants. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T00:20:17Z (GMT). No. of bitstreams: 1 ntu-100-R98b47114-1.pdf: 4817726 bytes, checksum: 32b28f8a17a857fe30586b63fd4860fe (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 口試委員會審定書 I
謝誌 II ABSTRACT III 中文摘要 IV Abbreviations V Index VI CONTENTS VII LIST OF TABLES AND FIGURES X Chapter 1 Introduction 1 1.1 Transgenic plant cell cultures 1 1.1.1 Plant cell cultures 1 1.1.2 Plant cells expressing system 2 1.2 Agrobacterium-mediated gene transformation 3 1.2.1 Agrobacterium species 3 1.2.2 Process of Agrobacterium-mediated transformation 4 1.2.3 Binary vector system 5 1.3 Hairy root cultures 5 1.4 Multiple genes co-expression 6 1.5 Reporter proteins 7 1.5.1 beta-glucuronidase 7 1.5.2 Green fluorescence protein 8 1.6 Research aim 8 Chapter 2 Materials and methods 10 2.1 Agrobacterium rhizogenes transformants 10 2.1.1 Binary vectors 10 2.1.2 Heat shock method for E.coli transformation 12 2.1.3 Electroporation for A. rhizogenes transformation 12 2.2 Induction of hairy roots 14 2.2.1 Sterilization of tobacco seeds 14 2.2.2 Induction of hairy roots 14 2.2.3 Liquid cultures of hairy roots 15 2.3 Foreign DNA confirmation 15 2.3.1 Extraction of genomic DNA 15 2.3.2 Confirmation of foreign genes with PCR 16 2.4 Functional GFP and GUS confirmation 17 2.4.1 Direct observation of GFP 17 2.4.2 GUS histochemical assay 18 2.5 Protein assay 18 2.5.1 Extraction of cytosol protein 18 2.5.2 Quantification of total protein 19 2.5.3 GUS activity assay 19 2.5.4 ELISA assay for GFP quantification 19 2.5.5 Statistic analysis 20 Chapter 3 Results 21 3.1 Infection of tobacco 21 3.1.1 Colony PCR of A. rhizogenes 21 3.1.2 Induction rate 21 3.2 PCR confirmation of transgenic hairy roots 22 3.3 The expression of reporter genes in transgenic hairy roots 23 3.3.1 Growth rate of hairy roots 23 3.3.2 Expression of GUS and GFP 24 3.3.3 Quantification of GUS and GFP 24 Chapter 4 Discussion 25 Tables and Figures 29 References 46 Appendix 52 | |
dc.language.iso | en | |
dc.title | 共同表現GUS與GFP於菸草毛狀根 | zh_TW |
dc.title | Co-expression of GUS and GFP in hairy roots of Nicotiana tabacum Wisconsin 38 | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 吳定峰,楊健志,洪傳揚 | |
dc.subject.keyword | 根毛農桿菌,菸草毛狀根,GFP,GUS,共轉形, | zh_TW |
dc.subject.keyword | A. rhizogenes,tobacco hairy root,GFP,GUS,co-transformation, | en |
dc.relation.page | 55 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-08-15 | |
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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