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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李平篤 | |
| dc.contributor.author | Shi-Yi Chuang | en |
| dc.contributor.author | 莊緒怡 | zh_TW |
| dc.date.accessioned | 2021-06-13T16:26:58Z | - |
| dc.date.available | 2005-07-28 | |
| dc.date.copyright | 2005-07-28 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-07-14 | |
| dc.identifier.citation | Amir J, Preiss J. 1982. Kinetic characterization of spinach leaf sucrose -phosphate synthase. Plant Physiol 69﹕1027-1030
Babb VM., Haigler CH. 2001. Sucrose phosphate synthase activity rises in correlation with high-rate cellulose synthesis in three heterotrophic systems. 2001. Plant Physiol. 127﹕1234-1242 Bird IF, Cornelius MJ, Keys AJ, Whittingham CP. 1974. Intracellular site of sucrose synthesis in leaves. Phytochemistry 13﹕59-64 Birnboim HC., Doly, J. 1979. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl Acids Res 7﹕1513-1523 Brierley RA. 1998. Secretion of recombinant human insulin-like growth factor I (IGF-1). Methods Mol. Biol. 103﹕149-177 Chiruvolu V., Cregg, JM., Meagher, MM. 1997. Recombinant protein production in an alcohol oxidase-defective strain of Pichia pastoris in fed-batch fermentations. Enzyme Microb. Technol. 21﹕277-283 Sugden C., Donaghy PG., Halford NG., Hardie DG. 1999. Two SNF1-related protein kinases from spinach leaf phosphorylate and inactivate 3-hydroxy-3-methylglutaryl-coenzyme A reductase, nitrate reductase, and sucrose phosphate synthase in vitro1. Plant Physiol 120﹕257-274 Couderc R., Baratti J. 1980. Oxidation of methanol by the yeast Pichia pastoris: purifcation and properties of alcohol oxidase. Agric. Biol. Chem. 44﹕2279-2289 Cregg JM, Madden KR. 1988. Development of the methylotrophic yeast, Pichia pastoris, as a host system for the production of foreign proteins. Dev Ind Microbiol 29﹕23-42 Doehlert DC, Huber SC. 1985. The role of sulfhydryl groups in the regulation of spinach leaf sucrose-phosphate synthase. Biochim. Biophys. Acta 830﹕267-273 Douglas P, Pigaglio E, Ferrer A, Halfold NG, MacKintosh C. 1997. Three spinach leaf nitrate reductase-3-hydroxy-3-methylglutaryl-CoA reductase kinases that are regulated by reversible phosphorylationand/or Ca2+ ions. Biochem J. 325﹕101-109 Ellis SB., Brust PF., Koutz PJ., Waters AF., Harpold MM., Gingeras TR. 1985. Isolation of alcohol oxidase and two other methanol regulatable genes from the yeast Pichia pastoris. Mol. Cell. Biol. 5﹕1111-1121 Faber KN, Harder W, Ab G, Veenhuis M. 1995. Review: methylotrophic yeasts as factories for the production of foreign proteins. Yeast 11﹕1331-1344. Goochee CF., Gramer MJ., Andersen DC., Bahr JB., Rasmussen JR. 1991. The oligosaccharides of glycoproteins: bioprocess factors affecting oligosaccharide structure and their effect on glycoprotein properties. Biotechnology (NY) 9﹕1347-1355 Harbron S, Foyer C, Walker D. 1981. The purification and properties of sucrose-phosphate synthase from spinach leaves: the involvement of this enzyme and fructose bisphosphatase in the regulation of sucrose biosynthesis. Arch Biochem Biophys 212﹕237-246 Hatch MD, Sacher JA, Glasziou KT. 1963. Sugar accumulation cycle in sugar cane. Studies on enzymes of the cycle. Plant Physiol 38﹕338-343 Hatzfeld DW., Moesinger E., 1995. Active-site-directed inhibition of sucrose phosphate synthase by Cibacron blue F3G-A and 1-deoxynojirimycin. Planta. 197﹕296-300 Hawker JS. 1969. Changes in the activities of enzymes concerned with sugar metabolism during the development of grape berries. Phytochemistry 8﹕9-17 Hawker JS. 1971. Enzymes concerned with sucrose synthesis and transformations in seeds of maize, broad been and castor bean. Phytochemistry 10﹕2313-2322 Hesse H., Sonnewald U., Willmitzer L. 1995. Cloning and expression analysis of sucrose-phosphate synthase from sugar beet (Beta vulgaris L.). Mol Gen Genet 247﹕515-520 Higgins DR., Busser K., Comiskey J., Whittier PS., Purcell TJ., Hoeffler JP. 1998. Small vectors for expression based on dominant drug resistance with direct multicopy selection. Methods Mol. Biol. 103﹕41-53 Huber SC, Huber JL, McMichael RW Jr. 1994. Control of plant enzyme activity by reversible protein phosphorylation. Int.Rev.Cytol. 149﹕47-98 Huber SC, Huber JL. 1991. In vitro phosphorylation and inactivation of spinach leaf sucrose-phosphate synthase by an endogenous protein kinase. Biochim. Biophys. Acta 1091﹕393-400 Huber SC, Huber JL. 1992. Role of sucrose-phosphate synthase in sucrose metabolism in leaves. Plant Physiol. 99﹕1275-78 Huber SC, Huber JL. 1996. Role and regulation of sucrose phosphate synthase in higher plants. Annu Rev Plant Physiol Plant Mol Biol 47﹕431-444 Huber SC, McMichael RW Jr, Huber JL, Bachmann M, Yamamoto YT, Conkling MA. 1995. Light regulation of sucrose synthesis: role of protein phosphorylation and possible involvement of cytosolic [Ca2+]. In Carbon Partitioning and Source-Sink Interactions in Plants, ed. MAMadore, W Lucas, 35-44. HuberJL., Huber SC., Nielsen TH. 1989a. Protein phosphorylation as a mechanism for regulation of spinach leaf sucrose-phosphate synthase activity. Arch. Biochem. Biophy. 270﹕681-690 Huber SC, Nielsen TH, Huber JLA, Pharr DM. 1989b. Variation among species in light activation of sucrose-phosphate synthase. Plant & Cell Physiology 30﹕ 277-285 Huber SC., Rufty TW., Keer PS. 1984. Effect of photoperiod on photosynthesis partitioning and diurnal rhythms in sucrose phosphate synthase activity in leaves of soybean and tobacco. Plant Physiol. 75﹕1080-1084 Cereghino JL., Cregg JM. 2000. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiology Reviews. 24﹕45-66 Klein RR., Crafts-Brandner SJ., Salvucci ME. 1993. Cloning and developmental expression of the sucrose-phosphate-synthase gene from spinach. Planta 190﹕498-510 Koch KE. 1996. Carbohydrate-modulated gene expression in plants. Annu Rev Plant Physiol Plant Mol Biol 47﹕509-540. Koutz P., Davis GR., Stillman C., Barringer K., Cregg J., Thill G. 1989. Structural comparison of the Pichia pastoris alcohol oxidase genes. Yeast 5 ﹕ 167-177 Lejeune P, Bernier G, Requier MC., kinet JM.1993. Sucrose increase during floral induction in the phloem sap collected at the apical part of the shoot of the long-day plant Sinapis alba L. Planta 190﹕71-74 Lunn JE, Ap RT. 1990a. Apparent equilibrium constant and mass-action ratio for sucrose-phosphate synthase in seeds of Pisum sativum. Biochemical Journal 267﹕739-743 Lunn JE, Ap RT. 1990b. Purification and properties of sucrose-phosphate synthase from seeds of Pisum sativum. Phytochemistry 29﹕1057-1064 McMichael RW Jr., Bachman M., Huber SC. 1995. Spinach leaf sucrose -phosphate synthase and nitrate reductase are phosphorylated/inactivated by multiple protein kinase in vitro. Plant Physiol 108﹕1077-1082 McMichael RW Jr., Klein RR., Salvucci ME., Huber SC. 1993. Identification of the major regulatory phosphorylation site in sucrose phosphate synthase. Arch. Biochem. Biophys. 307﹕248–52 Milyaeva EL., Komarova EN. 1996. Changes in the sugar content in stem apices of the short-day plant Perilla nankinensis at floral transition. Russ J Plant Physiol 43﹕149-154 Montesino R., Garcia R., Quintero O., Cremata JA. 1998. Variation in N-linked oligosaccharide structures on heterologous proteins secreted by the methylotrophic yeast Pichia pastoris. Protein Expr. Purif. 14﹕197-207 Moorhead G, Douglas P, Cotelle V, Harthill J, Morrice N, Meek S, Deiting U, Stitt M, Scarabel M, Aitken A, MacKintosh C. 1999. Phosphorylation–dependent interactions between enzymes of plant metabolism and 14-3-3 proteins. Plant J. 18﹕1–12 Muslin AJ, Tanner JW, Allen PM, Shaw AS. 1996. Interaction of 14-3-3 with signaling proteins is mediated by the recognition of phosphoserine. Cell. 84﹕889-97. Ogata K., Nishikawa H., Ohsugi M. 1969. A yeast capable of utilizing methanol. Agric. Biol. Chem. 33 : 1519-1520 Oliyai C, Patel JP., Carr L., Borchardt RT. 1994. Chemical pathways of peptide degradation. VII. Solid state chemical instability of an aspartyl residue in a model hexapeptide. Pharm Res. 11 : 901-908 Patrick SM., Fazenda ML., McNeil B., Harvey LM. 2005. Review:Heterologous protein production using the Pichia pastoris expression system. Yeast 22﹕249-270. Quick P, Siegl G, Neuhaus E, Feil R, Stitt M. 1989. Short-term water stress leads to a stimulation of sucrose synthesis by activating sucrose-phosphate synthase. Planta 177﹕535-546 Reimholz R., Geigenberger P., Stitt M. 1994. Sucrose phosphate synthase is regulated, via metabolites and protein phosphorylation in potato tubers, in a manner analogous to the enzyme in leaves. Planta 1992﹕480-88 Rorem ES., Walker HG., McMichael RW. 1960. Biosynthesis of sucrose and sucrose-phosphate by sugar beet leaf extracts. Plant Physiol. 35﹕269-273 Salvucci ME., Drake RR. 1990. Purification and photoaffinity labeling of sucrose phosphate synthase from spinach leaves. Arch.Biochem.Biophys. 281﹕212-218 Salvucci ME., Klein RR. 1993. Identification of the uridine-binding domain of sucrose-phosphate synthase: expression of a region of the protein that photoaffinity labels with 5-azidouridine diphosphate-glucose. Plant Physiol. 102﹕529–536 Scorer CA., Clare JJ., McCombie WR., Romanos MA., Sreekrishna K. 1994. Rapid selection using G418 of high copy number transformants of Pichia pastoris for high-level foreign gene expression. Biotechnology (NY) 12﹕ 181-184. Sheen J. 1990. Metabolic repression of transcription in higher plants. The Plant Cell 2﹕1027-1038. Siegl G., Stitt M. 1990. Partial purification of two forms of spinach sucrose -phosphate synthase which differ in their kinetic properties. Plant Sci. 66 : 205-210 Slabnik E, Frydman RB, Cardini CE. 1968. Some properties of potato tuber UDPG:D-fructose-2-glucosyltransferase (E.C. 2.4.1.14) and UDPG:D-fructose -6-phosphate-2-glucosyltransferase (E.C. 2.4.1.13). Plant Physiol. 43﹕1063 -1068 Sonnewald U., Basner A. 1993. EMBLData Library, Accession No. S34172 Stitt M, Huber SC, Kerr P. 1987. Control of photosynthetic sucrose formation. In Biochemistry of Plants, ed. MD Hatch, NK Boardman, 8﹕327-409. New York: Academic Sugiharto B, Sakakibara H, Saumadi, Sugiyama T. 1997. Differential expression of two genes for sucrose-phosphate synthase in sugarcane: molecular cloning of the cDNAs and comparative analysis of gene expression. Plant Cell Physiol. 38﹕961-965 Toroser D., Huber SC. 1997. Protein phosphorylation as a mechanism for osmotic-stress activation of sucrose-phosphate synthase in spinach leaves. Plant Physiol. 114﹕947-955 Toroser D., Athwal GS., Huber SC. 1998. Site-specifc regulatory interaction between spinach leaf sucrose-phosphate synthase and 14-3-3 proteins. FEBS Letters 435﹕110-114 Toroser D., Huber SC., 1998. 3-Hydroxy-3-methylglutaryl-coenzyme A reductase kinase and sucrose-phosphate synthase kinase activities in cauliflower florets: Ca2+-dependence and substrate specificities. Arch Biochem Biophys. 355﹕291-300 Tschopp JF., Brust PF., Cregg JM., Stillman CA., Gingeras TR. 1987. Expression of the LacZ gene from two methanol-regulated promoters in Pichia pastoris. Nucleic Acids Res. 15﹕3859-3876. Tschopp JF., Sverlow G., Kosson R., Craig W., Grinna L. 1987. High level secretion of glycosylated invertase in the methylotrophic yeast, Pichia pastoris. Bio/Technology 5﹕1305-1308 Valdez-Alarcon JJ., Ferrando M., Salerno G., Jimenez-Moraila B., Herrera-Estrella L. 1996. Characterization of a rice sucrose-phosphate synthase -encoding gene. Gene 170﹕217-222 Van Den Hazel HB., Kielland-Brandt MC., Winther JR. 1996. Review: biosynthesis and function of yeast vacuolar proteases.Yeast. 12(1)﹕1-16 Weiner H., Weiner H., Stitt M. 1993. Sucrose-phosphate synthase phosphatase, a type 2A protein phosphatase, changes its sensitivity towards inhibition by inorganic phosphate in spinach leaves. FEBS Lett. 333﹕159–64 Worrell AC., Bruneau JM., Summerfelt K., Boersig M., Voelker TA. 1991. Expression of a maize sucrose phosphate synthase in tomato alters leaf carbohydrate partitioning. Plant Cell 3﹕1121-1130 陳勁中 (2001) 甘藷葉蔗糖磷酯合成酶之生化學研究。碩士論文，國立台灣大學農業化學研究所。 林秋宏 (2001) 綠竹筍蔗糖合成酶cDNA之選殖與檢定。碩士論文，國立台灣大學農業化學研究所。 曾靖涵 (2002) 綠竹筍蔗糖磷酯合成酶之生化學研究。碩士論文，國立台灣大學農業化學研究所。 李木和 (2004) 光照與滲透逆境下水稻蔗糖磷酯合成酶性質鑑定與免疫組織定位。博士論文，國立台灣大學微生物與生化學研究所。 洪加正 (2004) 綠竹筍蔗糖磷酯合成酶cDNA之選殖與檢定。碩士論文，國立台灣大學微生物與生化學研究所。 鄭傑洋 (2005) 綠竹筍苯丙胺酸脫氨裂解酶在酵母菌中之表現與檢定。碩士論文，國立台灣大學微生物與生化學研究所。 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/38142 | - |
| dc.description.abstract | 蔗糖磷酯合成酶 (sucrose phosphate synthase，簡稱SPS) 為高等植物調控蔗糖生合成之重要酵素。本論文目的，即希望以具真核細胞轉譯後修飾作用之表現系統，表現綠竹筍蔗糖磷酯合成酶(BOSPS)。
首先，依據BOSPS cDNA 之5‘及3’ 端序列，設計含特定限制酶 (NotⅠ及ApaⅠ) 切位之引子，利用聚合酶鏈鎖反應 (PCR)，即得兩端分別具 NotⅠ 及 ApaⅠ 切位之BOSPS cDNA；將此cDNA 接入T載體yT&A，轉形至大腸桿菌 (JM109) 中保存。續以限制酶NotⅠ及ApaⅠ將其自T載體切下，與經相同限制酶處理之pPICZ A表現載體接合(pPBOSPS)後，轉形至大腸桿菌 (JM109) 進行定序分析；待序列確認無誤，以限制酶 SacⅠ將此重組表現載體切成線狀，轉形至Pichia pastoris 中 (X-33)。 確定酶母菌轉形株表現型及最適表現時間點後，即可以甲醇誘導BOSPS大量表現；繼則以10~30% PEG沈澱及鎳離子親和層析管柱純化之，所得酵素則可分別進行各項生化性質分析。 | zh_TW |
| dc.description.abstract | Sucrose phosphate synthase (SPS) is an important regulatory enzyme of sucorse biogenic reaction in plants. The purpose of this study is to express sucrose phosphate synthase from shoots of Bamboo (BOSPS) by eukaryotic expression system with posttranslational modification.
At first, forward (with restriction site NotⅠ) and reverse (with restriction site ApaⅠ) primers were designed according to the both terminal sequences of BOSPS cDNA. Then, BOSPS cDNA containing these restriction sites was amplified by PCR and cloned into T vector (yT&A). Both of the recombinant plasmid pTBOSPS and pPICZ A vector were digested by restriction enzymes ApaⅠand NotⅠ sequentially. Therefore, BOSPS cDNA can be cloned into pPICZ A vector (pPBOSPS) by the terminal complementary cohesive end. Being confirmed the correct nucleotide sequence by sequencing, recombinant plasmid pPBOSPS could be linearized and transformed into Pichia pastoris (X-33). Before scale-up of BOSPS expression induced by the addition of methanol, determining the Mut phenotype and optimal time post-induction to harvest. Finally, expressed BOSPS was purified by a procedure involving precipitation with 10~30% PEG and affinity chromatography, and analyzed for its characterization. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T16:26:58Z (GMT). No. of bitstreams: 1 ntu-94-R91623612-1.pdf: 803022 bytes, checksum: f02e33938443624cfaead3fc695db672 (MD5) Previous issue date: 2005 | en |
| dc.description.tableofcontents | 中文摘要………………………………………………………………Ⅰ
Abstract………………………………………………………………Ⅱ 縮寫表…………………………………………………………………Ⅲ 第一章 緒論……………………………………………………………1 第一節 蔗糖……………………………………………………………1 第二節 蔗糖磷酯合成酶………………………………………………2 第三節 Pichia pastoris表現系統…………………………………9 第四節 實驗緣起……………………………………………………14 第二章 材料與方法……………………………………………………16 第一節 實驗材料、藥品與儀器………………………………………16 1.1 實驗材料…………………………………………………………16 1.2 實驗藥品與儀器…………………………………………………16 第二節 一般實驗法……………………………………………… 17 2.1 瓊脂糖膠體電泳…………………………………………………17 2.2 質體DNA小量.………………………………………………… 18 2.3 瓊脂糖膠體中DNA片段之純化………………………………20 2.4 電穿孔法competent cell之製備………………………………20 2.5 綠竹筍蔗糖磷酯合成酶酵素活性測定………………………23 2.6 蛋白質定量法……………………………………………………24 2.7 蛋白質電泳檢定系統……………………………………………26 2.8 蛋白質轉印法…………………………………………………31 2.9 免疫染色法………………………………………………………31 第三節 Pichia pastoris表現系統之建立…………………………33 3.1 聚合酶鏈鎖反應 (PCR)……………………………………35 3.2 yT&A 重組載體之建構、轉形與檢定…………………………32 3.3 pPICZ A重組載體之建構、轉形與檢定……………………39 第四節 綠竹筍蔗糖磷酯合成酶之大量表現與純化…………………44 4.1 Pichia pastoris轉形株表現型之檢定………………………44 4.2 綠竹筍蔗糖磷酯合成酶最適誘導時間點之探討………………45 4.3 綠竹筍蔗糖磷酯合成酶之大量表現……………………………47 4.4 純化經表現之綠竹筍蔗糖磷酯合成酶…………………………47 第五節 經表現之綠竹筍蔗糖磷酯合成酶生化性質探討……………50 5.1 最適反應溫度……………………………………………………50 5.2 最適反應pH值…………………………………………………50 5.3 Glc 6-P對酵素活性之影響……………………………………51 5.4 Pi對酵素活性之影響………………………………………52 5.5 不同金屬離子對酵素活性之影響……………………………53 第三章 結果與討論…………………………………………………54 第一節 Pichia pastoris表現系統之建立…………………………54 1.1 聚合酶鏈鎖反應 (PCR) ………………………………………54 1.2 yT&A重組載體之建構、轉形與檢定……………………………56 1.3 pPICZ A重組載體之建構、轉形與檢定……………………58 第二節 綠竹筍蔗糖磷酯合成酶之大量表現與純化…………………59 2.1 Pichia pastoris轉形株表現型之檢定………………………59 2.2 綠竹筍蔗糖磷酯合成酶最適誘導時間點之探討……………60 2.3 綠竹筍蔗糖磷酯合成酶之大量表現與純化…………………61 第三節 經表現之綠竹筍蔗糖磷酯合成酶生化性質探討……………62 3.1 最適反應溫度.…………………………………………………62 3.2 最適反應pH值………………………………………………… 62 3.3 Glc 6-P對酵素活性之影響…………………………………62 3.4 Pi對酵素活性之影響……………………………………………63 3.5 不同金屬離子對酵素活性之影響………………………………63 第四章 結論與展望…………………………………………………73 附錄……………………………………………………………………75 參考文獻………………………………………………………………81 | |
| dc.language.iso | zh-TW | |
| dc.subject | 蔗糖磷酯合成酶 | zh_TW |
| dc.subject | 綠竹筍 | zh_TW |
| dc.subject | 酵母菌 | zh_TW |
| dc.subject | Sucrose phosphate synthase | en |
| dc.subject | Bamboo(Bambusa oldhamii) | en |
| dc.subject | Yeast(Pichia pastoris) | en |
| dc.title | 綠竹筍蔗糖磷酯合成酶在酵母菌中之表現 | zh_TW |
| dc.title | Expression of Sucrose Phosphate Synthase from Shoots of Bamboo (Bambusa oldhamii)in Yeast (Pichia pastoris) | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 林耀輝,林棋財,楊健志 | |
| dc.subject.keyword | 蔗糖磷酯合成酶,綠竹筍,酵母菌, | zh_TW |
| dc.subject.keyword | Sucrose phosphate synthase,Bamboo(Bambusa oldhamii),Yeast(Pichia pastoris), | en |
| dc.relation.page | 87 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2005-07-15 | |
| 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 | |
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