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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.author | Tun-Hsun Kuo | en |
dc.contributor.author | 郭暾珣 | zh_TW |
dc.date.accessioned | 2021-07-01T08:12:29Z | - |
dc.date.available | 2021-07-01T08:12:29Z | - |
dc.date.issued | 2002 | |
dc.identifier.citation | ( PARTⅠ)
[l] Muta, T., and Iwanaga, S. (1996) The role of hemolymph coagulation in innate immunity. Curr. Opin. Immunol. 8, 41-47. [2] Iwanaga, S., Kawabata, and S., Muta, T. (1998) New types of clotting factors and defense molecules found in horseshoe crab hemolymph: their structures and functions. J. Biochem, 123, 1-15. [3] Fortes-Dias, C. L., Minetti, C. A. S., Lin, Y., and Liu, T.-Y. (1993) Agglutination activity of Limulus polyphemus coagulogen following limited proteolysis. Comp. Biochem. Physiol. 105, 79-85. [4] Minetti, C. S. A., Lin, Y, Cislo, T., and Liu, Y. (1991) Purification and characterization of an endotoxin-binding protein with protease inhibitory activity from Limulus amebocytes. J. Biol. Chem. 266, 20773-20780. [5] Liu, T., Lin, Y., Cislo, T., Minetti, C. A. S. A., Baba, J. M. K., and Liu, T. Y.(1991) Limunectin. A phosphocholine-binding protein from Limulus amebocytes with adhesion-promoting properties. J. Biol. Chem. 266, 14813-14821. [6] Kawabata, S., and Iwanaga, S. (1999) Role of lectins in the innate immunity of horseshoe crab. Dev. Comp. Immunol. 23,391-400. [7] Gokudan, S., Muta, T., TSuda, R., Koori, K., Kawahara, T., Seki, N., Mizunoe, Y., W. S. N., Iwanaga, S., and Kawabata, S. (1999) Horseshoe crab acetyl group-recognizing lectins involved in innate immunity are structurally related to fibrinogen. Proc. Acad Sci. U. S. A. 96, 10086-10091. [8] Chiou, S. -T., Chen, Y. -W., Chen, S. -C., Chao, C. -F., and Liu, T. Y. (2000) Isolation and characterization of proteins that bind to galactose, lipopolysaccharide of Escherichia coli, and protein A of Staphylococcus aureus from the hemolymph of Tachypleus tridentatus. J. Biol. Chem. 275,1630-1634. [9] Chen, S. -C., Yen, C. -H., Yeh, M. S., Huang, C. J., and Liu, T. Y. (2000) Biochemical properties and cDNA cloning of two new lectins from the plasma of Tachypleus tridentatus: Tachypleus plasma lectin 1 and 2. J. Biol. Chem. 276, 9631-9639. [10] Saito, T., Kawabata, S., Hirata, M., and Iwanaga, S. (1995) A novel type of limulus lectin-L6. Purification, primary structure, and antibacterial activity. J. Biol. Chem. 270, 14493-14499. [11] Nagai, T., Kawabata, S., Shishikura, F., and Sugita, H. (1999) Purification, characterization, and amino acid sequence of an embryonic lectin in perivitelline fluid of the horseshoe crab. J. Biol. Chem. 274, 37673-37678. [12] Hong, T. H., Chen, S. T., Tang, T. K., Wang, S. C., and Chang, T. H. (1989) The production of polyclonal and monoclonal antibodies in mice using novel immunization methods. J. Immunol. Methods 21, 151-157. [13] Nguyen, N. H., Suzuki, A., Cheng, S. M., Zon, G., and Liu, T. Y. (1986) Isolation and characterization of Limulus C-reactive protein genes. J Biol. Chem. 261, 10450-10455. [14] Okino, N., Kawabata, S. I., Saito, T., Hirata, M., Takagi, T., and Iwanaga, S. (1995) Purification, characterization, and cDNA cloning of a 27-kDa lectin (L10) from horseshoe crab hemocytes. J Biol Chem. 270, 31008-31015. [15] Inamori, K. I., Saito, T., Iwaki, D., Nagira, T., Iwanaga, S., Arisaka, F., and Kawabata, S. (1999) A newly identified horseshoe crab lectin with specificity for blood group A antigen recognizes specific O-antigens of bacterial lipopolysaccharides. J. Biol. Chem. 274, 3272-3278. [16] Nagai, T., Kawabata, S., Shishikura, F., and Sugita, H. (1999) Purification, characterization, and amino acid sequence of an embryonic lectin in perivitelline fluid of the horseshoe crab. J. Biol. Chem. 274, 37673-37678. [17] Iwanaga, S. (2002) The molecular baiss of innate immunity in the horseshe crab. Curr. Opin. Immunol. 14,87-95. [18] Beisel H. G., Kawabata, S., Iwanaga, S., Huber, R., and Bode, W. (1999) Tachylectin-2: crysatl structure of a specific GlcNAc/GalNAc binding lectin involved in the innate immunity host defense of the Japanese horseshoe crab Tachypleus tridentatus. EMBO J. 18, 2313-2322. [19] Inamori, K., Saito, T., Iwaki, D., Nagira, T., Iwanaga, S., Arisaka, F., and Kawabata, S. (2001) Horseshoe crab hemocyte-derived lectin recognizing specific O-antigens of lipopolysaccharides. Adv. Exp. Med. Biol. 484,185-190. [20] Saito, T., Hatada, M., Iwanaga, S., and Kawabata, S. (1997) A newly identified horseshoe crab lectin with binding specificity to O-antigen of bacterial lipopolysaccharides. J. Biol. Chem. 272, 30703-30708. [21] Iwaki, D., Osaki, T., Mizunoe, Y., Wai, S. N. Iwanaga, S., and Kawabata, S. (1999) Functional and diversities of C-reactive proteins present in horseshoe crab hemolymph plasma. Eur. J. Biochem. 264,314-326. [22] Kairies, N., Beisel, H., Fuentes-Prior, P., Tsuda, R., Muta, T., Iwanaga, S., Bode, W., Huber, R., and Kawabata, S. (2001) The 2.0-A crystal of tachylectin 5A provides evidence for the common origin of the innate immunity and the blood coagulation systems. Proc. Natl. Acad Sci. 98, 13519-13524. ( PARTⅡ) [1] Ogura, K., Koyama, T. and Sagami, H (1997) Polyprenyl diphosphate synthases. Subcellular Biochem. 28,57-87. [2] Ogura, K. and Koyama, T. (1998) Enzymatic aspects of isoprenoid chain elongation. Chemical Reviews 98,1263-1276. [3] Shimizu, N., Koyama, T., and Ogura, K. (1998) Molecular cloning, expression, and purification of undecaprenyl diphosphate synthase. No sequence similarity between E- and Z-prenyl diphosphate synthases. J. Biol. Chem. 273, 19476-19481. [4] Apfel, C. M., Takacs, B., Fountoulakis, M., Stieger, M., and Keck, W. (1999) Use of genomics to identify bacterial undecaprenyl pyrophosphate synthetase: cloning, expression, and characterization of the essential gene. J. Bacteriol.181,483-492. [5] Fujihashi, M., Zhang, Y. -W., Higuchi, Y., Li, X.-Y., Koyama, T., and Miki, K. (2001) Crystal structure of cis-prenyl chain elongating enzyme, undecaprenyl diphosphate synthase. Proc. Acad. Sci. 98,4337-4342. [6] Baba, T., and Allen, C. M. (1980) Prenyl transferase from Micrococcus luteus: characterization of undecaprenyl pyrophosphate synthase. Arch. Biochem. 200,474-484. [7] Allen, C. M., Keenan, M.V., and Sack, J. (1976) Lactobacillus plantarum undecaprenyl pyrophosphate synthase purification and reaction requirement. Arch. Biochem. Biophys. 175, 236-248. [8] Keenan, M.V., and Allen, C. M. (1974) Characterization of undecaprenyl pyrophosphate synthase from Lactobacillus plantarum. Arch. Biochem. Biophys. 161,375-383. [9] Allen, C. M., and Muth, J. D. (1977) Lipid activation of undecaprenyl pyrophosphate synthase from Lactobacillus plantarum. Biochemistry 16, 2908-2915. [10] Keenan, M. V., and Allen, C. M. (1974) Phospholipid activation of Lactobacillus plantarum undecaprenyl pyrophosphate synthetase. Biochem. Biophys.Res. Commun. 61,338-342. [11] Allen, C.M. (1985) Purification and characterization of undecaprenyl pyrophosphates synthase. Methode in Enzymol. 110,281-299. [12] Pan, J. J., Chiu, S. T., and Liang, P.H. (2000) Product distribution and pre-steady-state kinetic analysis of E. coli undecaprenyl pyrophospate synthase. Biochemistry 39,10936-10942. [13] Ohnuma, S. -I., Koyama, T., and Ogura, K. (1991) Purification of solanesyl-diphosphate synthase from Micrococcus luteus. A new class of prenyltransferase. J. Biol. Chem. 266, 23706-23713. [14] Fujisaki, S., Nishino, T., and Katsuki, H. (1986) Isoprenoid synthesis in Escherichia coli. Separation and partial purification of four enzymes involved in the synthesis. J. Biochem. 99, 1327-1337. [15] Asai, K. -I., Fujisaki, S., Nishimura, Y, Nishino, T., Okada, K., Nakagawa, T., Kawamukai, M., and Matsuda, H. (1994) The identification of Escherichia coli ispB (cel) gene encoding the octaprenyl diphosphate. Biochem. Biophys. Res. Commun. 202,340-345. [16] Okada, K., Suzuki, K., Kamiya, Y., Zhu, X., Fujisaki, S., Nishimura, Nishino, T., Nakagawa, T., Kawamukai, M., and Matsuda, H. (1996) Polyprenyl diphosphate synthase essentially defines the length of the side chain of ubiquinone. Biochim. Biophys. Acta 1302,217-223. [17] Okada, K., Minehira, M., Zhu, X., Suzuki, K., Nakagawa, T., Matsuda, H., and Kawamukai, M. (1997) The ispB gene encoding octaprenyl diphosphate synthase is essential for growth of Escherichia coli. J. Bacteriol. 179,3058-3060. [18] Kainou, T., Okada, K., Suzuki, K., Nakagawa, T., Matsuda, H., and Kawamukai, M. (2001) Dimer formation of octaprenyl-diphosphate synthase (IspB) is essential for chain length determination of ubiquinone. J. Biol. Chem. 276,7876-7883. [19] Adams, M. W. W. (1993) Enzymes and proteins from organisms that grow near and above 100 degrees celcius. Annu. Rev. Microbiol. 47,627-658. [20] Day, M. W., Hsu, B. T., Joshua-Tor, L., Park, J. B., Zhou, Z. H., Adams, M. W. and Rees, D. C. (1992) X-ray crystal structures of the oxidized and reduced forms of the rubredoxin from them marine hyperthermophilic archaebacterium Pyrococcus furiosus. Protein Sci. 1, 1494-1507. [21] Huber, R., Langworthy, T. A., Konig, H., Thomm, M., Woese, C. R., Sleytr, U. B., and Stetter, K.O.(1986) Thermotoga maritima sp. nov. represents a new genus of unique extremely thermophilic eubacteria growing up to 90℃. Arch Microbiology 144,324-333. [22] Collin, M. D., Jones, D. (1981) Distribution of isoprenoid quinone structural types in bacteria and their taxonomic implication. Microbiol. Rev. 45, 316-354. [23] Teclebrhan, H., Olsson, J., Swiezewska, E., and Dallner, G. (1993) Biosynthesis of the side of chain of ubiquinone: trans-prenyltransferase in rat liver microsomes. J. Biol. Chem. 268,23081-23086. [24] Ernster, L., and Dallner, G. (1995) Biochemical, physiological and medical aspects of ubiquinone function. Biochim. Biophys. Acta 1271,195-204. [25] Fujii, H., Koyama, T., and Ogura, K. (1982) Efficient enzymatic hydrolysis of polyprenyl pyrophosphate. Biochimica et Biophysica Acta 712,716-718. [26] Barshop, B. A., Wrenn, R. F., and Frieden, C. (1983) Analysis of numerical methods for computer simulation of kinetic processes: development of KINSIM-a flexible, portable system. Anal. Biochem. 130,134-145. [27] Davison, P. F. (1968) Proteins in denaturing solvents: gel exclusion studies. Science 161, 906-907. [28] Sreerama, N., and Woody, R. W. (1993) A self-consistent method for the analysis of protein secondary from circular dichroism. Anal. Biochem. 209,32-44. [29] Eftink, M. R. (1995) Use of multiple spectroscopic methods to monitor equilibrium unfolding of proteins. Methods Enzymol. 259,487-512. [30] Pan, J. J., Chiu, S. T., and Liang, P. H. (2000) Product distribution and pre-steady-state kinetic analysis of E. coli undecaprenyl pyrophosphate synthase. Biochemistry 39,10936-10942. [31] Ko, T. P., Chen, Y. K., Robinson, H., Tsai, P. C., Gao, Y. G., Chen, A. P. -C., A. H. -J., and Liang, P. H. (2001) Mechanism of the product chain length determination and the role of a flexible loop in undecaprenyl pyrophosphate synthase catalysis. J. Biol. Chem. 276, 47474-47486. [32] Ohnuma, S. -I., Koyama, T., and Ogura, K. (1992) Chain length distribution of the products formed in solanesyl diphosphate synthase reaction. J. Biochem. (Tokyo) 112,743-749. [33] Ashby, M. N., and Edwards, P. A. (1990) Elucidation of the deficiency in two yeast coenzyme Q mutants. Characterization of the structural gene encoding hexaprenyl pyrophosphates synthetase. J. Biol. Chem. 265, 13157-13164. [34] Koyama, T., Obata, S., Osabe, M., Takeshita, A., Yokoyama, K., Uchita, M. Nishino, T., and Ogura, K. (1993) Thermostable farnesyl diphosphates synthase of Bacillus stearothermophilus: molecular cloning sequence determination, overproduction, and purification. J. Biochem. (Tokyo) 113,355-363. [35] Chen, A., Kroon, P. A., and Poulter, C. D. (1994) Isoprenyl diphosphate synthases: protein comparisons, a phylogenetic tree, and predictions of secondarys structure. Protein Sci. 3, 600-607. [36?Tarshis, L. C., Yan, M., Poulter, C. D., and Sacchettini, J. C. (1994) Crystal of recombinant farnesyl diphosphate synthase at 2.6 A resolution. Biochemistry 33, 10871-10877. [37] Marrero, P. F., Poulter, C. D., and Edwards, P. A. (1992) Effects of site-directed mutagenesis of the highly conserved aspartate residues in domain Ⅱof the farnesyl diphosphate synthase activity. J. Biol. Chem. 267, 21873-21878. [38] Joly, A., and Edwards, P. A. (1993) Effect of site-directed mutagenesis of conserved aspartate and arginine residues upon farnesyl diphosphate synthase activity. J. Biol. Chem. 268, 26983-26989. [39] Song, L., and Poulter, C. D. (1994) Yeast farnesyl-diphosphate synthase: site-directed genesis of residues in highly conserved prenyltransferase domainsⅠand Ⅱ. Proc. Natl. Acad Sci. U.S. A. 91, 3044-3048. [40] Koyama, T., Tajima, M., Sano, H., Doi, T., Koike-Takeshita, A. Obata, S. Nishino, T., and Ogura, K. (1996) Identification of significant residues in the binding site of Bacillus stearothermophilus farnesyl diphosphate synthase. Biochemistry 35,9533-9538. [41] Koyama, T., Saito, K. Ogura, K., Obata, S., and Takeshita, A. (1994) Thermostable farnesyl diphosphates synthase of Bacillus stearothermophilus: crystallization and site-directed mutagenesis. J. Chem. 72, 75-79. [42?Tarshis, L. C., Proteau, P. J., Kellogg, B. A., Sacchettini, J. C., and Poulter, C. D. (1996) Regulation of product chain length by isoprenyl diphosphate synthases. Proc. Natl. Acad. Sci. U.S.A. 93,15018-15023. [43] Poulter, C. D., and Rilling, H. C. (1976) Prenyltransferase: the mechanism of the reaction. Biochemistry 9, 1079-83. [44] Pan, J. J., Kuo, T. H., Chen, Y. K., Yang, L. W., and Liang, P. H. (2002) Insight into the activation mechanism of E. coli octaprenyl pyrophosphate synthase derived from pre-steady-state kinetic analysis. Biochim. Biophys. Acta, 1594,64-73. [45] Altschul, S. F., Madden, T. L., Schaffe, A. A., Zhang, J., Zhang, Z., Miller, W., and Lipman, D. J. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search program. Nucleic acid Res. 25, 3389-3402. [46] Chen, Y. H., Chen, A. P. -C., Chen, C. T., Wang, A. H. -J., and Liang, P. H. (2002) Probing the conformational change of Escherichia coli undecaprenyl pyrophosphate synthase during catalysis using an inhibitor and tryptophan mutants. J. Biol. Chem. 277,7369-7376. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75275 | - |
dc.description.abstract | 第一部份:鱟血清外源凝集素-1和-2在天賦免疫系統抵禦上,扮演和細菌結合的功能。先前劉德勇教授實驗室從台灣鱟血清中分離出二種外源凝集素[Chiou, S-T., Chen, Y.-W., Chen, S.-C., Chao, C.-F. and Liu, T.-Y.(2000) J. Biol. Chem.275,1630-1634],及克隆它們的基因發現其為分子量26-kDa 和12-Da的醣蛋白[Chen, S.-C., Yen, C.-H., and Liu, T.-Y,(2001)J.Biol.Chem.276,9631-9639]。描述在本論文中,我使用酵母菌表達系統使這些蛋白分泌至培養基而能簡易純化及鑑定重組蛋白特性。表達之蛋白由於不同程度的醣化而在SDS-PAGE上出現二個band,其可被切醣酵素切成和不具醣化的突變蛋白在SDS-PAGE上相同位置。使用ELISA研究其ligand結合專一性顯示鱟血清外源凝集素-1和N-acetyl醣類結合而鱟血清外源凝集素-2辨識細菌表面脂多醣之O-antigen部份。當外源凝集素-1的醣化序列的Asn被Asp所取代,即失去細菌結合能力表示醣化對鱟血清外源凝集素-1是必需的。另一方面、外源凝集素-2醣化序列的N3D突變並不影響其功能。就蛋白組成而言、分子間雙硫鍵所形成的dimer對於鱟血清外源凝集素-2是必需的且脂多醣造成其多元體形成。
第二部份:八異戊二烯焦磷酸合成酵素(OPPs)催化五個異戊二烯焦磷酸(IPP)和法呢基焦磷酸(FPP)反應生成產物八異戊二烯焦磷酸,其可形成ubiquinone的支鏈。由於長鏈產物離開酵素不易、此類酵素常需要加入detergent或其他因數維持其最佳活性。先前發現加Triton可將十一異戊二烯焦磷酸合成酵素的速率決定部驟從產物離開轉變成異戊二烯焦磷酸化學反應[Pan, J.J., Chiou, S.T., and Liang, R.H.(2000) Biochemistry 39,10936-109421]。為瞭解八異戊二烯焦磷酸合成酵素之活性激發機制,我們測量酵素單一循環反應(single-turnover)發現異戊二烯焦磷酸的化學反應速率常數為2 s-1、100倍大於steady stat測得的反應速率常數0.02 s-1, 細菌細胞內高分子量蛋白和Triton皆可將反應速率常數增加約三倍、但不足以造成100倍效果。產物以burst方式形成亦證明在有或無Triton狀況,產物離開很慢為速率決定部驟。此部份已發表[Pan, J.J., Kuo, T.H., Chen, Y.K., Yang, L.W., and Liang, P. H. (2002) Insight into the activation mechanism of E. coli octaprenyl pyrophosphates synthase derived from pre-steady-state kinetic analysis. Biochim. Biophys. Acta 1594, 64-73]. 我亦表達、純化及進行動力學研究對耐熱菌的八異戊二烯焦磷酸合成酵素。此酵素在室溫下的活性為0.005 s-l,且活性隨溫度昇高而呈exponential增加。和大腸桿菌不同的是耐熱菌的八異戊二烯焦磷酸合成酵素的化學反應為速率決定部驟。反應產物為40個碳的八異戊二烯焦磷酸且其轉變成45個碳速率幾乎可以忽略。在有10 ?M OPPs-FPP複合物和1?M IPP單一循環反應(single-turnover)狀況下,只有C20而非由大腸桿菌酵素所產生的C20-C40。這些數據顯示大腸桿菌和耐熱菌的八異戊二烯焦磷酸合成酵素動力學的差異,反應出耐熱菌酵素較高的活性區域穩定度。 | zh_TW |
dc.description.abstract | PartⅠ: Tachypleus Plasma Lectin-1 and -2 (TPL-1 and -2), which bind bacteria for innate host defense, were previously isolated from the hemolymph of Taiwanese Tachypleus tridentatus [Chiou, S.-T., Chen, Y.-W., Chen, S. -C., Chao, C.-F. and Liu, T.-Y.(2000)J.Biol.Chem.275,1630-1634] and their genes encoding a 26-kDa and a 12-kDa glycol-proteins were cloned in Dr. Liu's laboratory [Chen, S. -C., Yen, C. -H., and Liu, T .-Y.,(2001)J.Biol.Chem.276,9631-9639]. In the present study, TPL-1 and -2 produced using yeast and the recombinant proteins secreted into medium were purified and characterized. The purified lectins show two bands on the SDS-PAGE as resulted from heterogeneous glycosylation and the endo-H glycosidase treatment could generate a single band, which migrates on the SDS-PAGE at the same position as non-glycosylated mutant protein. Ligand specificity examination using ELISA assay reveals that TPL-1 interacts with N-acetyl saccharides and TPL-2 recognizes O-antigen of bacterial lipopolysaccharides. The glyco moiety of the TPL-1 is essential for its function as the substitution of Asn in the N-glycosylation site with Asp abolishes the ligand binding affinity. On the other hand, N3D mutant TPL-2 retains bacterial binding activity. The dimer formation by intermolecular disulfide linkage is essential for TPL-2 activity and the LPS induces its oligomerization.
PartⅡ.Octaprenyl pyrophosphate synthase (OPPs) catalyzes the sequential condensation of five molecules of isopentenyl pyrophosphate with farnesyl pyrophosphate (FPP) to generate all-trans C40-octaprenyl pyrophosphate, which constitutes the side chain of ubiquinone. Due to the slow product release, a long-chain polyprenyl pyrophosphate synthase often requires detergent or other factor for optimal activity. Our previous studies in examining the activity enhancement of E. coli undecaprenyl pyrophosphate synthase have demonstrated a switch of rate determining step from product release to IPP condensation reaction in the presence of Triton [Pan, J.J., Chiou, S.T., and Liang, P.H.(2000) Biochemistry 39,10936-109421]. In order to understand the mechanism of enzyme activation for E.coli OPPs, a single-turnover reaction was performed and the measured IPP condensation rate (2 s-l) was 100 times larger than the stead-state rate (0.02 s-1). The high molecular weight fractions and Triton could accelerate the steady-stat rate by 3-fold (0.06 s-1) but insufficient to cause full activation (100-fold). A burst product formation was observed in enzyme multiple turnovers indicating a slow product release. This part has been published [Pan, J.J., Kuo, T.H., Chen, Y.K., Nhng, L. W., and Liang, P.H. (2002) Insight into the activation mechanism of E. coli octaprenyl proophosphate synthase derived from pre-steady-state kinetic analysis. Biochim. Biophys. Acta 1594,64-73]. Moreover, a putative gene encoding for OPPs from T. maritima, an anaerobic and thermophilic bacterium,was expressed, purified and its kinetic pathway was determined. The enzyme activity at 25℃ is 0.005 s-1 under steady-state condition and exponentially increased with elevated temperature. In contrast to E. coli OPPs, IPP condensation rather than product release is rate limiting in enzyme reaction. The product of chain elongation catalyzed by T maritima OPPs is C40 and the rate of its conversion to C45 is negligible. Under single-turnover condition with 10 ?M OPPs. FPP complex and 1 pM IPP, only the C20 was formed rather than C20-C40 observed for E.coli enzyme. These data reveal the differences in kinetic properties of OPPs from T maritima and E.coli, reminiscent of lower enzyme activity at room temperature, higher product specificity, higher thermal stability and lower structure flexibility for the thermophilic enzyllle. | en |
dc.description.provenance | Made available in DSpace on 2021-07-01T08:12:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2002 | en |
dc.description.tableofcontents | ABSTRACT ………………………………………………………………………………4 ABBREVIAT1ONS…………………………………………………………………………10 INTRODUCTION (PARTⅠ)………………………………………………………………12 MATERIALS AND METHODS (PARTⅠ)……………………………………………………15 RESULTS (PARTⅠ)………………………………………………………………………25 DISCUSSION (RARTⅠ) …………………………………………………………………27 TABLES (PARTⅠ)………………………………………………………………………31 FIGURES (PARTⅠ)………………………………………………………………………34 REFERENCES (PARTⅠ)…………………………………………………………………40 INTRODUCTION (PARTⅡ)………………………………………………………………44 MATERIALS AND METHODS (PARTⅡ)…………………………………………………49 RESULTS (PARTⅡ) ……………………………………………………………………60 DISCUSSION(PERTⅡ)…………………………………………………………………68 TABLES (PARTⅡ)………………………………………………………………………74 SCHEMES (PARTⅡ)………………………………………………………………………76 FIGURES (PARTⅡ)………………………………………………………………………79 SUPPORTING INFORMATION ……………………………………………………………88 REFERENCES (PARTⅡ) …………………………………………………………………97 | |
dc.language.iso | zh-TW | |
dc.title | 第一部份:鱟血清中細菌結合蛋白表達及特性研究 第二部份:大腸桿菌及耐熱菌之八異戊二磷酸合成酵素之反應機制及動力學研究 | zh_TW |
dc.title | PartⅠ: Expression and Characterization of Tachyplcus Plasma Lectin-1 and -2 from Taiwanese Tachypleus tridentatus PartⅡ: Mechanistic and Kinetic Studies on Recombinant Octaprenyl Pyrophosphate Synthase from E. coli and T.maritima | en |
dc.date.schoolyear | 90-2 | |
dc.description.degree | 碩士 | |
dc.relation.page | 104 | |
dc.rights.note | 未授權 | |
dc.contributor.author-dept | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科學研究所 | zh_TW |
顯示於系所單位: | 生化科學研究所 |
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