請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25276完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊健志(Chien-Chih Yang) | |
| dc.contributor.author | Chao-Li Huang | en |
| dc.contributor.author | 黃兆立 | zh_TW |
| dc.date.accessioned | 2021-06-08T06:07:26Z | - |
| dc.date.copyright | 2007-07-23 | |
| dc.date.issued | 2007 | |
| dc.date.submitted | 2007-07-17 | |
| dc.identifier.citation | Alper, T., Haig, D.A., and Clarke, M.C. (1966). The exceptionally small size of the scrapie agent. Biochem. Biophys. Res. Commun. 22, 278-284.
Alper, T., Cramp, W.A., Haig, D.A., and Clarke, M.C. (1967). Does the agent of scrapie replicate without nucleic acid? Nature 214, 764-766. Anderson, R.M., Donnelly, C.A., Ferguson, N.M., Woolhouse, M.E.J., Watt, C.J., Udy, H.J., MaWhinney, S., Dunstan, S.P., Southwood, T.R.E., Wilesmith, J.W., Ryan, J.B.M., Hoinville, L.J., Hillerton, J.E., Austin, A.R., and Wells, G.A.H. (1996). Transmission dynamics and epidemiology of BSE in British cattle. Nature 382, 779-788. Basler, K., Oesch, B., Scott, M., Westaway, D., Walchli, M., Groth, D.F., McKinley, M.P., Prusiner, S.B., and Weissmann, C. (1986). Scrapie and cellular PrP isoforms are encoded by the same chromosomal gene. Cell 46, 417-428. Bocharova, O.V., Breydo, L., Parfenov, A.S., Salnikov, V.V., and Baskakov, I.V. (2005). In vitro Conversion of Full-length Mammalian Prion Protein Produces Amyloid Form with Physical Properties of PrPSc. J. Mol. Biol. 346, 645-659. Bolton, D.C., McKinley, M.P., and Prusiner, S.B. (1982). Identification of a protein that purifies with the scrapie prion. Science 218, 1309-1311. Borchelt, D., Scott, M., Taraboulos, A., Stahl, N., and Prusiner, S. (1990). Scrapie and cellular prion proteins differ in their kinetics of synthesis and topology in cultured cells. J. Cell Biol. 110, 743-752. Bosques, C.J., and Imperiali, B. (2003). The interplay of glycosylation and disulfide formation influences fibrillization in a prion protein fragment. Proc. Natl. Acad. Sci. USA 100, 7593-7598. Bounhar, Y., Zhang, Y., Goodyer, C.G., and LeBlanc, A. (2001). Prion Protein Protects Human Neurons against Bax-mediated Apoptosis. J. Biol. Chem. 276, 39145-39149. Breydo, L., Sun, Y., Makarava, N., Lee, C.I., Novitskaia, V., Bocharova, O., Kao, J.P., and Baskakov, I.V. (2007). Nonpolar substitution at the C-terminus of the prion protein, a mimic of the glycosylphosphatidylinositol anchor, partially impairs amyloid fibril formation. Biochemistry 46, 852-861. Brown, D.R., Hafiz, F., Glasssmith, L.L., Wong, B.S., Jones, I.M., Clive, C., and Haswell, S.J. (2000). Consequences of manganese replacement of copper for prion protein function and proteinase resistance. The EMBO J. 19, 1180-1186. Brown, P., Cervenakova, L., McShane, L., Goldfarb, L.G., Bishop, K., Bastian, F., Kirkpatrick, J., Piccardo, P., Ghetti, B., and Gajdusek, D.C. (1998). Creutzfeldt-Jakob disease in a husband and wife. Neurology 50, 684-688. Bruce, M.E., Will, R.G., Ironside, J.W., McConnell, I., Drummond, D., Suttie, A., McCardle, L., Chree, A., Hope, J., Birkett, C., Cousens, S., Fraser, H., and Bostock, C.J. (1997). Transmissions to mice indicate that `new variant' CJD is caused by the BSE agent. Nature 389, 498-501. Bueler, H., Fischer, M., Lang, Y., Bluethmann, H., Lipp, H.-P., DeArmond, S.J., Prusiner, S.B., Aguet, M., and Weissmann, C. (1992). Normal development and behaviour of mice lacking the neuronal cell-surface PrP protein. Nature 356, 577-582. Capellari, S., Cardone, F., Notari, S., Schinina, M.E., Maras, B., Sita, D., Baruzzi, A., Pocchiari, M., and Parchi, P. (2005). Creutzfeldt-Jakob disease associated with the R208H mutation in the prion protein gene. Neurology 64, 905-907. Carrell, R.W., and Lomas, D.A. (1997). Conformational disease. The Lancet 350, 134-138. Caughey, B., and Raymond, G.J. (1991). The scrapie-associated form of PrP is made from a cell surface precursor that is both protease- and phospholipase-sensitive. J. Biol. Chem. 266, 18217-18223. Chesebro, B., Race, R., Wehrly, K., Nishio, J., Bloom, M., Lechner, D., Bergstrom, S., Robbins, K., Mayer, L., Keith, J.M., Garon, C., and Haase, A. (1985). Identification of scrapie prion protein-specific mRNA in scrapie-infected and uninfected brain. Nature 315, 331-333. Chesebro, B., Trifilo, M., Race, R., Meade-White, K., Teng, C., LaCasse, R., Raymond, L., Favara, C., Baron, G., Priola, S., Caughey, B., Masliah, E., and Oldstone, M. (2005). Anchorless prion protein results in infectious amyloid disease without clinical scrapie. Science 308, 1435-1439. Chiarini, L.B., Freitas, A.R., Zanata, S.M., Brentani, R.R., Martins, V.R., and Linden, R. (2002). Cellular prion protein transduces neuroprotective signals. The EMBO J. 21, 3317-3326. Collee, J.G., and Bradley, R. (1997). BSE: a decade on--part I. The Lancet 349, 636-641. Collinge, J. (2001). Prion Diseases of Humans and Animals: Their Causes and Molecular Basis. Annu. Rev. Neurosci. 24, 519-550. Coustou, V., Deleu, C., Saupe, S., and Begueret, J. (1997). The protein product of the het-s heterokaryon incompatibility gene of the fungus Podospora anserina behaves as a prion analog. Proc. Natl. Acad. Sci. USA 94, 9773-9778. David A Hilton, A.C.G.L.C.P.E.L.M.D.R.M.P.D.H.J.W.I. (2004). Prevalence of lymphoreticular prion protein accumulation in UK tissue samples. J. Pathol. 203, 733-739. Davis-Salinas, J., and Van Nostrand, W.E. (1995). Amyloid beta-Protein Aggregation Nullifies Its Pathologic Properties in Cultured Cerebrovascular Smooth Muscle Cells. J. Biol. Chem. 270, 20887-20890. Derkatch, I.L., Chernoff, Y.O., Kushnirov, V.V., Inge-Vechtomov, S.G., and Liebman, S.W. (1996). Genesis and Variability of [PS1] Prion Factors in Saccharomyces cerevisiae. Genetics 144, 1375-1386. Donne, D.G., Viles, J.H., Groth, D., Mehlhorn, I., James, T.L., Cohen, F.E., Prusiner, S.B., Wright, P.E., and Dyson, H.J. (1997). Structure of the recombinant full-length hamster prion protein PrP(29-231): The N terminus is highly flexible. Proc. Natl. Acad. Sci. USA 94, 13452-13457. Enari, M., Flechsig, E., and Weissmann, C. (2001). Scrapie prion protein accumulation by scrapie-infected neuroblastoma cells abrogated by exposure to a prion protein antibody. Proc. Natl. Acad. Sci. USA 98, 9295-9299. Gabriel, J., Oesch, B., Kretzschmar, H., Scott, M., and Prusiner, S.B. (1992). Molecular Cloning of a Candidate Chicken Prion Protein. Proc. Natl. Acad. Sci. USA 89, 9097-9101. Gajdusek, D. (1996). Infectious amyloids: subacute spongiform encephalopathies as transmissible cerebral amyloidoses. In: BN Field, PM Knipe and PM Howley et al., Editors, Field's Virology (3rd edn.), Lippincott-Raven, Philadelphia 2851–2900. Gajdusek, D.C., and Zigas, V. (1957). Degenerative disease of the central nervous system in New Guinea; the endemic occurrence of kuru in the native population. N. Engl. J. Med. 257, 974-978. Glenner, G.G., Eanes, E.D., Bladen, H.A., Linke, R.P., and Termine, J.D. (1974). Beta-pleated sheet fibrils. A comparison of native amyloid with synthetic protein fibrils. J. Histochem. Cytochem. 22, 1141-1158. Goldfarb, L.G., Petersen, R.B., Tabaton, M., Brown, P., LeBlanc, A.C., Montagna, P., Cortelli, P., Julien, J., Vital, C., Pendelbury, W.W., and et, a. (1992). Fatal familial insomnia and familial Creutzfeldt-Jakob disease: disease phenotype determined by a DNA polymorphism. Science 258, 806-808. Hecker, R., Taraboulos, A., Scott, M., Pan, K.M., Yang, S.L., Torchia, M., Jendroska, K., DeArmond, S.J., and Prusiner, S.B. (1992). Replication of distinct scrapie prion isolates is region specific in brains of transgenic mice and hamsters. Genes Dev. 6, 1213-1228. Hill, A.F., Desbruslais, M., Joiner, S., Sidle, K.C.L., Gowland, I., Collinge, J., Doey, L.J., and Lantos, P. (1997). The same prion strain causes vCJD and BSE. Nature 389, 448-450. Hilton, D.A., Fathers, E., Edwards, P., Ironside, J.W., and Zajicek, J. (1998). Prion immunoreactivity in appendix before clinical onset of variant Creutzfeldt-Jakob disease. The Lancet 352, 703-704. Hsiao, K., Baker, H.F., Crow, T.J., Poulter, M., Owen, F., Terwilliger, J.D., Westaway, D., Ott, J., and Prusiner, S.B. (1989). Linkage of a prion protein missense variant to Gerstmann-Straussler syndrome. Nature 338, 342-345. Huang, Z., Prusiner, S.B., and Cohen, F.E. (1995). Scrapie prions: a three-dimensional model of an infectious fragment. Fold. Des. 1, 13-19. Jackson, G.S., Murray, I., Hosszu, L.L.P., Gibbs, N., Waltho, J.P., Clarke, A.R., and Collinge, J. (2001). Location and properties of metal-binding sites on the human prion protein. Proc. Natl. Acad. Sci. USA 98, 8531-8535. Johnson, R.T. (2005). Prion diseases. Lancet Neurol. 4, 635-642. Jones, C.E., Klewpatinond, M., Abdelraheim, S.R., Brown, D.R., and Viles, J.H. (2005). Probing Copper2+ Binding to the Prion Protein Using Diamagnetic Nickel2+ and 1H NMR: The Unstructured N terminus Facilitates the Coordination of Six Copper2+ Ions at Physiological Concentrations. J. Mol. Biol. 346, 1393-1407. Kaneko, K., Zulianello, L., Scott, M., Cooper, C.M., Wallace, A.C., James, T.L., Cohen, F.E., and Prusiner, S.B. (1997). Evidence for protein X binding to a discontinuous epitope on the cellular prion protein during scrapie prion propagation. Proc. Natl. Acad. Sci. USA 94, 10069-10074. Katscher, F. (1998). It's Jakob's disease, not Creutzfeldt's. Nature 393, 11-11. Laurent, M. (1996). Prion diseases and the 'protein only' hypothesis: a theoretical dynamic study. Biochem. J. 318 ( Pt 1), 35-39. Lee, I.Y., Westaway, D., Smit, A.F., Wang, K., Seto, J., Chen, L., Acharya, C., Ankener, M., Baskin, D., Cooper, C., Yao, H., Prusiner, S.B., and Hood, L.E. (1998). Complete genomic sequence and analysis of the prion protein gene region from three mammalian species. Genome Res. 8, 1022-1037. Lee, L.Y.-L., and Chen, R.P.-Y. (2007). Quantifying the Sequence-Dependent Species Barrier between Hamster and Mouse Prions. J. Am. Chem. Soc. 129, 1644-1652. Lehmann, S., and Harris, D.A. (1997). Blockade of Glycosylation Promotes Acquistion of Scrapie-like Properties by the Prion Protein in Cultured Cells. J. Biol. Chem. 272, 21479-21487. Lewis, P.A., Properzi, F., Prodromidou, K., Clarke, A.R., Collinge, J., and Jackson, G.S. (2006). Removal of the glycosylphosphatidylinositol anchor from PrP(Sc) by cathepsin D does not reduce prion infectivity. Biochem. J. 395, 443-448. Li, A., Dong, J., and Harris, D.A. (2004). Cell Surface Expression of the Prion Protein in Yeast Does Not Alter Copper Utilization Phenotypes. J. Biol. Chem. 279, 29469-29477. Li, G., and Bolton, D.C. (1997). A novel hamster prion protein mRNA contains an extra exon: increased expression in scrapie. Brain Res. 751, 265-274. Locht, C., Chesebro, B., Race, R., and Keith, J.M. (1986). Molecular Cloning and Complete Sequence of Prion Protein cDNA from Mouse Brain Infected with the Scrapie Agent. Proc. Natl. Acad. Sci. USA 83, 6372-6376. Mange, A., Milhavet, O., Umlauf, D., Harris, D., and Lehmann, S. (2002). PrP-dependent cell adhesion in N2a neuroblastoma cells. FEBS Lett. 514, 159-162. Manson, J.C., Clarke, A.R., Hooper, M.L., Aitchison, L., McConnell, I., and Hope, J. (1994). 129/Ola mice carrying a null mutation in PrP that abolishes mRNA production are developmentally normal. Mol. Neurobiol. 8, 121-127. Manuelidis, L., Yu, Z.-X., Barquero, N., and Mullins, B. (2007). Cells infected with scrapie and Creutzfeldt-Jakob disease agents produce intracellular 25-nm virus-like particles. Proc. Natl. Acad. Sci. USA 104, 1965-1970. Marsh, R.F., and Kimberlin, R.H. (1975). COMPARISON OF SCRAPIE AND TRANSMISSIBLE MINK ENCEPHALOPATHY IN HAMSTERS .2. CLINICAL SIGNS, PATHOLOGY, AND PATHOGENESIS. J. Infect. Dis. 131, 104-110. Medori, R., Montagna, P., Tritschler, H.J., LeBlanc, A., Cortelli, P., Tinuper, P., Lugaresi, E., and Gambetti, P. (1992). Fatal familial insomnia: A second kindred with mutation of prion protein gene at codon 178. Neurology 42, 669-. Meyer, R.K., McKinley, M.P., Bowman, K.A., Braunfeld, M.B., Barry, R.A., and Prusiner, S.B. (1986). Separation and Properties of Cellular and Scrapie Prion Proteins. Proc. Natl. Acad. Sci. USA 83, 2310-2314. Montagna, P., Gambetti, P., Cortelli, P., and Lugaresi, E. (2003). Familial and sporadic fatal insomnia. The Lancet Neurol. 2, 167-176. Oesch, B., Westaway, D., Walchli, M., McKinley, M.P., Kent, S.B.H., Aebersold, R., Barry, R.A., Tempst, P., Teplow, D.B., Hood, L.E., Prusiner, S.B., and Weissmann, C. (1985). A cellular gene encodes scrapie PrP 27-30 protein. Cell 40, 735-746. Oltersdorf, T., Fritz, L.C., Schenk, D.B., Lieberburg, I., Johnson-Wood, K.L., Beattie, E.C., Ward, P.J., Blacher, R.W., Dovey, H.F., and Sinha, S. (1989). The secreted form of the Alzheimer's amyloid precursor protein with the Kunitz domain is protease nexin-II. Nature 341, 144-147. Pattison, I.H., and Jones, K.M. (1968). Modification of a strain of mouse-adapted scrapie by passage through rats. Res. .Vet. Sci. 9, 408-410. Paul Brown, C.J.G.J.P.R.-J.D.M.A.M.P.S.A.B.L.G.G.D.C.G. (1994). Human spongiform encephalopathy: The national institutes of health series of 300 cases of experimentally transmitted disease. Ann. Neurol. 35, 513-529. Pauly, P.C., and Harris, D.A. (1998). Copper Stimulates Endocytosis of the Prion Protein. J. Biol. Chem. 273, 33107-33110. Peden, A.H., and Ironside, J.W. (2004). Review: pathology of variant Creutzfeldt-Jakob disease. Folia neuropathologica / Association of Polish Neuropathologists and Medical Research Centre, Polish Academy of Sciences 42 Suppl A, 85-91. Prusiner, S.B. (1982). Novel proteinaceous infectious particles cause scrapie. Science 216, 136-144. Prusiner, S.B., Groth, D.F., Bolton, D.C., Kent, S.B., and Hood, L.E. (1984). Purification and structural studies of a major scrapie prion protein. Cell 38, 127-134. Prusiner, S.B., Bolton, D.C., Groth, D.F., Bowman, K.A., Cochran, S.P., and McKinley, M.P. (1982). Further purification and characterization of scrapie prions. Biochemistry 21, 6942-6950. R. G. Will, M.Z.G.E.S.M.A.M.J.W.I.S.N.C.J.M.K.E.A.J.E. (2000). Diagnosis of new variant Creutzfeldt-Jakob disease. Ann. Neurol. 47, 575-582. Riek, R., Hornemann, S., Wider, G., Glockshuber, R., and Wuthrich, K. (1997). NMR characterization of the full-length recombinant murine prion protein, mPrP(23-231). FEBS Lett. 413, 282-288. Riek, R., Hornemann, S., Wider, G., Billeter, M., Glockshuber, R., and Wuthrich, K. (1996). NMR structure of the mouse prion protein domain PrP(121-231). Nature 382, 180-182. Rogers, M., Taraboulos, A., Scott, M., Groth, D., and Prusiner, S.B. (1990). Intracellular accumulation of the cellular prion protein after mutagenesis of its Asnlinked glycosylation sites. Glycobiology 1, 101-109. Saeki, K., Matsumoto, Y., Hirota, Y., Matsumoto, Y., and Onodera, T. (1996). Three-exon structure of the gene encoding the rat prion protein and its expression in tissues. Virus Genes 12, 15-20. Schwartz, M. (2003). How the cows turned mad. University of California Press, Berkerly. Scott, M., Foster, D., Mirenda, C., Serban, D., Coufal, F., Walchli, M., Torchia, M., Groth, D., Carlson, G., DeArmond, S.J., and et al. (1989). Transgenic mice expressing hamster prion protein produce species-specific scrapie infectivity and amyloid plaques. Cell 59, 847-857. Scott, M.R., Groth, D., Tatzelt, J., Torchia, M., Tremblay, P., DeArmond, S.J., and Prusiner, S.B. (1997). Propagation of prion strains through specific conformers of the prion protein. J. Virol. 71, 9032-9044. Sigurdsson, B. (1954). Rida, a chronic encephalitis of sheep with general remarks on infections which develop slowly and some of their special characteristics. Vet. J. 110, 341. Sparkes, R.S., Simon, M., Cohn, V.H., Fournier, R.E.K., Lem, J., Klisak, I., Heinzmann, C., Blatt, C., Lucero, M., Mohandas, T., DeArmond, S.J., Westaway, D., Prusiner, S.B., and Weiner, L.P. (1986). Assignment of the Human and Mouse Prion Protein Genes to Homologous Chromosomes. Proc. Natl. Acad. Sci. USA 83, 7358-7362. Stahl, N., Baldwin, M.A., Burlingame, A.L., and Prusiner, S.B. (1990). Identification of glycoinositol phospholipid-linked and truncated forms of the scrapie prion protein. Biochemistry 29, 8879-8884. Stockel, J., Safar, J., Wallace, A.C., Cohen, F.E., and Prusiner, S.B. (1998). Prion Protein Selectively Binds Copper(II) Ions. Biochemistry 37, 7185-7193. Sun, Y., Breydo, L., Makarava, N., Yang, Q., Bocharova, O.V., and Baskakov, I.V. (2007). Site-specific Conformational Studies of Prion Protein (PrP) Amyloid Fibrils Revealed Two Cooperative Folding Domains within Amyloid Structure. J. Biol. Chem. 282, 9090-9097. Telling, G.C., Scott, M., Mastrianni, J., Gabizon, R., Torchia, M., Cohen, F.E., DeArmond, S.J., and Prusiner, S.B. (1995). Prion propagation in mice expressing human and chimeric PrP transgenes implicates the interaction of cellular PrP with another protein. Cell 83, 79-90. Telling, G.C., Scott, M., Hsiao, K.K., Foster, D., Yang, S.L., Torchia, M., Sidle, K.C., Collinge, J., DeArmond, S.J., and Prusiner, S.B. (1994). Transmission of Creutzfeldt-Jakob disease from humans to transgenic mice expressing chimeric human-mouse prion protein. Proc. Natl. Acad. Sci. USA 91, 9936-9940. Treiber, C., Simons, A., and Multhaup, G. (2006). Effect of copper and manganese on the de novo generation of protease-resistant prion protein in yeast cells. Biochemistry 45, 6674-6680. Westaway, D., Cooper, C., Turner, S., Costa, M.D., Carlson, G.A., and Prusiner, S.B. (1994a). Structure and Polymorphism of the Mouse Prion Protein Gene. Proc. Natl. Acad. Sci. USA 91, 6418-6422. Westaway, D., Zuliani, V., Cooper, C.M., Costa, M.D., Neuman, S., and Jenny, A.L. (1994b). Homozygosity for prion protein alleles encoding glutamine-171 renders sheep susceptible to natural scrapie. Genes Dev. 8, 959-969. Whittal, R.M., Ball, H.L., Cohen, F.E., Burlingame, A.L., Prusiner, S.B., and Baldwin, M.A. (2000). Copper binding to octarepeat peptides of the prion protein monitored by mass spectrometry. Protein Sci. 9, 332-343. Wickner, R.B. (1994). Evidence for a prion analog in S. cerevisiae: the [URE3] non-Mendelian genetic element as an altered URE2 protein. Science 264, 566. Will, R.G., and Matthews, W.B. (1984). A retrospective study of Creutzfeldt-Jakob disease in England and Wales 1970-79. I: Clinical features. J. Neurol. Neurosurg. Psychiatry. 47, 134-140. Wong, B.-S., Pan, T., Liu, T., Li, R., Petersen, R.B., Jones, I.M., Gambetti, P., Brown, D.R., and Sy, M.-S. (2000). Prion Disease: A Loss of Antioxidant Function? Biochem. Biophys. Res. Commun. 275, 249-252. Yoshimoto, J., Iinuma, T., Ishiguro, N., Horiuchi, M., Imamura, M., and Shinagawa, M. (1992). Comparative sequence analysis and expression of bovine PrP gene in mouse L-929 cells. Virus Genes 6, 343-356. Zuegg, J., and Gready, J.E. (2000). Molecular dynamics simulation of human prion protein including both N-linked oligosaccharides and the GPI anchor. Glycobiology 10, 959-974. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25276 | - |
| dc.description.abstract | 普昂蛋白質 (prion protein, PrP) 是一個藉 GPI-anchor 連接在膜上的醣蛋白質，由正常構形 (PrPC) 轉變為致病構形 (PrPSc) 的過程，被廣泛接受為引起普昂疾病的原因，但這個轉變過程仍不清楚，包括醣基化的影響以及物種屏障 (species barrier) 的成因，都未成定論。我們試著利用可進行轉譯後修飾作用的表現系統，例如進行醣基化，來表現重組 PrP。因此，本論文在 Escherichia coli 與 Pichia pastoris 中建立了重組小鼠 PrP 的表現系統。將 E. coli 中純化得到的重組蛋白質進行澱粉樣蛋白質纖維之培養。以 thioflavin T (ThT) 呈色法監控纖維的形成，再藉由穿透式電子顯微鏡 (Transmission electron microscope, TEM) 觀察纖維的形狀。基於合成胜肽系統推導出關於物種屏障的理論，而建構出三個突變蛋白質。依照金倉鼠 PrP 的序列，將小鼠重組 PrP 序列中 108, 111, 138 這三個位置的胺基酸換成 methionine。重組小鼠 PrP(23-230) 蛋白質的表現系統也同時建構在 P. pastoris 中，結果顯示由酵母菌產生的重組 PrP 也會存在不可溶的蛋白質中。 | zh_TW |
| dc.description.abstract | Prion protein (PrP) is a glycoprotein attached to plasma membrane via GPI-anchor. It is believed that the conversion from normal cellular PrP (PrPC) to pathogenic scrapie PrP (PrPSc) results in prion diseases. However, many issues still remained elusive, e.g. the effects of glycosylation and the factors that contributed to species barrier. We tried to express recombinant PrP in an expression system which was able to provide post-translational modifications, such as glycosylation. Therefore, recombinant wild type mouse PrP expression in Escherichia coli and Pichia pastoris were established. Recombinant wild type PrP purified from E. coli was used to form amyloid fibrils. The formation of the amyloid fibril was monitored by thioflavin T (ThT) assay. Transmission electron microscopy (TEM) was used to examine the morphology of the fibrils. Based on the theory for species barrier derived from synthetic peptide system established in previous study, three PrP(23-230) mutants were generated in an attempt to study the theory. The amino acids at positions of 108, 111, 138 in mouse PrP sequence were altered to methionine according to the PrP sequence of golden Syrian hamster. Recombinant mouse PrP(23-230) was also constructed for the expression in P. pastoris. It appeared that the recombinant PrP produced by yeast was also accumulated in the insoluble fraction. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T06:07:26Z (GMT). No. of bitstreams: 1 ntu-96-R94b47216-1.pdf: 4179460 bytes, checksum: 80dda1598b5305ab56c72dda1fe5b4ae (MD5) Previous issue date: 2007 | en |
| dc.description.tableofcontents | 目錄................................................................................................................................…………….......I
縮寫表.................................................................................................................................................IV 摘要.......................................................................................................................................................V Abstract..............................................................................................................................................VI 第一章 緒論 1.1 普昂疾病的簡介 ......................................................................................................................1 1.1.1 羊搔癢症 (scrapie) ...................................................................................................1 1.1.2 庫賈氏症 (CJD, Creutzfeldt-Jakob disease) .....................................................2 1.1.3 古魯症 (kuru) .............................................................................................................3 1.1.4 致命性家族失眠症 (FFI, fatal familial insomnia) ...........................................4 1.1.5 狂牛症 (BSE, bovine spongiform encephalopathy, mad cow disease) 與 vCJD (variant CJD) ..............................................................................................4 1.2 普昂 (prion) 簡介與其近代研究 ......................................................................................6 1.3 普昂蛋白質之簡介 .................................................................................................................8 1.3.1 普昂蛋白質的發現 ...................................................................................................8 1.3.2 轉譯出普昂蛋白質的基因 .....................................................................................9 1.3.3 小鼠普昂蛋白質之序列與功能性區塊 ..............................................................9 1.3.4 普昂蛋白質的生理功能 ........................................................................................10 1.3.5 普昂蛋白質的結構 .................................................................................................11 1.4 普昂疾病的物種屏障 (species barrier) .........................................................................12 1.5 轉譯後修飾作用與普昂蛋白質轉變為致病形態的關係 .........................................13 1.5.1 醣基化對普昂蛋白質的影響 ..............................................................................14 1.5.2 GPI-anchor 對普昂蛋白質的影響 ...................................................................15 1.6 澱粉樣蛋白質 (amyloid) 與相關疾病之簡介 ...........................................................16 1.7 研究動機與方向 ...................................................................................................................17 第二章 材料與方法 2.1 實驗材料 ................................................................................................................................20 2.1.1 載體 ............................................................................................................................20 2.1.2 菌株 ............................................................................................................................21 2.1.2.1 大腸桿菌 ...............................................................................................21 2.1.2.2 酵母菌 ...................................................................................................22 2.2 實驗藥品 ................................................................................................................................22 2.2.1 一般化學藥品 ..........................................................................................................22 2.2.2 酵素 ............................................................................................................................22 2.2.3 培養基 ........................................................................................................................23 2.3 儀器設備 ................................................................................................................................24 2.4 實驗方法 ................................................................................................................................25 2.4.1 DNA 相關之操作方法 .........................................................................................25 2.4.1.1 質體 DNA 小量製備 .......................................................................25 2.4.1.2 質體 DNA 中量製備 .......................................................................26 2.4.1.3 DNA 限制酶切 (Restriction Enzyme Digestion) ....................27 2.4.1.4 洋菜膠體電泳 (Agarose Gel Electrophoresis) .........................27 2.4.1.5 DNA 片段純化 (DNA Gel Extraction) ........................................27 2.4.1.6 DNA 片段接合 (Ligation) ..............................................................28 2.4.1.7 大腸桿菌化學法轉型 ........................................................................28 2.4.1.8 酵母菌 Pichia pastoris 之電穿孔 (Electroporation) 轉型 28 2.4.1.9 酵母菌染色體 DNA 之抽取 .........................................................29 2.4.1.10 聚合酶鏈鎖反應 (Polymerase Chain Reaction, PCR) ...........30 2.4.1.11 聚合酶鏈鎖反應產物純化 (PCR clean up) ................................31 2.4.1.12 QuikChange® 點突變技術 .............................................................31 2.4.2 蛋白質相關之操作方法 ........................................................................................32 2.4.2.1 重組蛋白質之最佳表現條件篩選 .................................................32 2.4.2.2 重組蛋白質之表現 ............................................................................33 2.4.2.3 E. coli 表現系統中 inclusion bodies 之純化 .........................35 2.4.2.4 P. pastoris 表現系統中非可溶性蛋白質之純化 ......................36 2.4.2.5 重組蛋白質之純化 ............................................................................37 2.4.2.6 蛋白質電泳檢定 .................................................................................39 2.4.2.7 蛋白質免疫轉印法 ............................................................................42 2.4.2.8 澱粉樣蛋白質纖維養成 ...................................................................44 2.4.2.9 以 thioflavin T 法偵測澱粉樣蛋白質纖維之生成 .................44 2.4.2.10 穿透式電子顯微技術 (Transmission Electron Microscopy, TEM) 樣本之製備 .............................................................................45 第三章 結果與討論 3.1 重組小鼠普昂蛋白質在大腸桿菌表現系統中之研究 .............................................46 3.1.1 進行點突變使重組蛋白質與 C 端標記接合 ................................................46 3.1.2 最適表現條件之探討 ............................................................................................47 3.1.3 重組小鼠普昂蛋白質之 inclusion body 的分離與純化 ..........................47 3.1.4 重組小鼠普昂蛋白質的純化 ..............................................................................48 3.1.5 重組小鼠普昂蛋白質之澱粉樣蛋白質纖維養成 .........................................50 3.1.6 對重組小鼠普昂蛋白質進行點突變以模仿金倉鼠普昂蛋白質 ..............50 3.1.7 各突變株之表現與 inclusion bodies 的純化 ..............................................51 3.1.8 各突變株之蛋白質純化 ........................................................................................52 3.1.9 以突變蛋白質 rPrP_LVM 進行澱粉樣蛋白質纖維養成 ...........................53 3.2 重組小鼠普昂蛋白質在酵母菌表現系統中之研究 ..................................................55 3.2.1 小鼠普昂蛋白質之基因序列選殖 .....................................................................55 3.2.2 載體的轉型與高抗性轉型株之篩選 ................................................................57 3.2.3 最適表現菌株及最適表現條件之探討 ............................................................59 3.2.4 鑑別在 Pichia pastoris 中表現的重組蛋白質是否可溶 ..........................60 第四章 結論與未來展望 4.1 結論 .........................................................................................................................................62 4.2 未來展望 ................................................................................................................................63 參考文獻 .........................................................................................................................................64 圖與表 ..............................................................................................................................................72 碩士論文口試問答摘要 ...........................................................................................................105 | |
| dc.language.iso | zh-TW | |
| dc.title | 重組小鼠普昂蛋白質表現系統之建構與研究 | zh_TW |
| dc.title | Construction and Studies of Recombinant Mouse Prion Protein Expression System | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 95-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 陳佩燁(Pei-Yeh Rita Chen) | |
| dc.contributor.oralexamcommittee | 蘇仲卿,李平篤 | |
| dc.subject.keyword | 普昂,小鼠, | zh_TW |
| dc.subject.keyword | prion protein,PrP,mouse, | en |
| dc.relation.page | 71 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2007-07-19 | |
| 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-96-1.pdf 未授權公開取用 | 4.08 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。