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  1. NTU Theses and Dissertations Repository
  2. 生命科學院
  3. 生化科學研究所
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75491
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dc.contributor.authorMING-SUN LIUen
dc.contributor.author劉明山zh_TW
dc.date.accessioned2021-07-01T08:13:29Z-
dc.date.available2021-07-01T08:13:29Z-
dc.date.issued1983
dc.identifier.citation1. Keller, D. F. (1971) G-6-PD Deficency.
2. Carson, P. E., Blanagan, C. L., Ickes, C. E. and Alving, A. S. (1956) Science 124, 484.
3. WHO Scientific Groun (1967) WHO Tech. Rep. Ser. 366, 1.
4. Harris, H. (1971) The Principle of Human Biochemical Genetics.
5. Kirkman, H. N. (1959) Nature 184, 1291.
6. Bonsignore, A., Cancedda, R., Lorenzoni, I., Cosulich, M. E., and De Flor, A. (1971) Biochem. Biophys. Res. Comms. 47, 94.
7. Wrigley, N. G., Heather, J. V. (1972) J. Mol. Biol. 68, 483.
8. Child, B., Zinkham, W., Brown, E. A., Kimbro, E. L., Torbet, J. V. (1958) Bull. Johns Hopkins Hosp. 102, 21.
9. Gaetani, G. D., Parker, J. C., Kirkman, H. N., (1974) Proc. Nat. Acad. Sci. USA 71, 3584.
10. Kirkman, H. N., Wilson, W. G., Clemons, E. H. (1980) J. Lab. Clin. Med. 95, 877.
11. Wilson, W. G., Kirkman, H. N., Clemons, F. H. (1980) J. Lab. Clin. Med. 95, 888.
12. Eggleston, L. V., Krebs, H. A. (1974) Biochem. J. 138, 425.
13. De Flora, A., Morelli, A., Benatti, U., Giuliano, F. and Molinari, P. (1974) Biochem. Biophys. Res. Comms. 60, 999.
14. Bonigno, A., Cancedda, R., Nicolini, A., Damiani, G., and De Flora, A. (1971) Arch. Biochem. Biophys. 147, 493.
15. Morelli, A., Benatti, U., Gaetani, G. F.and De Flora, A. (1978) Proc. Natl. Acad. Sci. USA 75, 1979.
16. Morelli. A., Benatti, U. (1974) Ital. J. Biochem. 23, 279.
17. Lee, C.-Y., Johansson, C. J. (1977) Anal. Biochem. 77, 90.
18. De Flora, A., Morelli, A.., Benatti, U. and Givliano, F. (1975) Arch. Biochem. Biophys. 169, 362.
19. Kahn, A., Dreyfus, J. C. (19711) Biochim. Biohys. Acta. 334, 257.
20. Morelli, A., De Flora, A. (1977) Arch. Biochem. Biophys. 179, 698.
21. Nakatsuji, T., Miwa, S. (1981) Anal. Biochem. 112, 52.
22. O’Farrell, P. H. (1975) J. Biol. Chem. 250 4007.
23. Kaloustian, V. M. D., Idriss-Daouk, S. H., Hallal, R. T., and Awdeh, Z. L. (1974) Biochem. Genet. 12, 51.
24. Hedrick, J. L. (1968) Arch. Biochem. Biophys. 126, 154.
25. Brodelius, P., Larason, P. O. and Mosbach, K. (1974) Eur. J. Biochem. 47, 81.
26. Larsson, P.-O., Mosbach, K. (1974) Methods Enzymol. 34, 235.
27. Lee, C.-Y., Langley, C. H., and Burkhart, J. (1978) Anal. Biochem. 86, 697.
28. Yorhida, A. (1966) J. Biol. Chem. 241, 4966.
29. Dc Flora, A., Giuliano, F. and Morelli, A. (1979) Ital. J. Biochem. 22, 258.
30. Hunter, L. (1980) Anal. Biochem. 101, 78.
31. Hunter, L. (1978) Anal. Biochem. 89, 279.
32. Kahn, A., Bertrand, O., Cottreau, D., Boivin, P. and Dreyfus, J. C. (1977) Biochem. Biophys. Res. Comms. 77, 65.
33. Kahler, S. G., Kirkman, H. N. (1983) J. Biol. Chem. 258, 717.
dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75491-
dc.description.abstract葡萄醣六磷酸去氫?(Glucose-6-phosphate dehydrogenase, G6PD)是人類紅血球維持正常生理代謝機能時不可缺少的酵素。
我們以N6-(aminohexyl) adenosine 2,5 biphosphate接於Sepharose 4B去做親和柱體(Affinity Chromatography)。可自人體紅血球分離得純質G6PD,以電泳鑑定其純度,亦證明此種分離方法十分良好。
我們於1M urea存在下,用粗製G6PD做等電荷焦點電泳(Isoelectric focusing, IEF),發現至少有七種以上的G6PD band出現。其等電點分別為pH 6.88, 6.79, 6.64, 6.5, 6.39, 6.19, 6.10。再將此七種G6PD band做disc electrophoresis,而發現每一 G6PD band又可分成二個成份,經鑑定而知其中之一分子量為96,000左右,可能是G6PD dimer,另一分子量為50,600-66,800,可能是monomer或是dimer分解物。
zh_TW
dc.description.abstractA non-equilibrium pH gradient electrophoresis conducted in 6'acrylamide gel in the presence of 1.0 M urea was used to separate isozymes of g1ucose-6-phosphate dehydrogenase in human erythrocytes. At least seven sharp bands were found to vary both in isoelectric point and in the relative intensity. Their isoelectric points, using pH 5-8 range ampholytes, were pH 6.88. for band 1, pH 6.79 for band 2, pH 6.64 for band 3, pH 6.5 for band 4, pH 6.39 for band 5, pH 6.19 for band 6, pH 6.1 for band 7. Each band was resolved further by a disc electrophoresis into two components, the slow(S) and the fast(f) components defined according to their mobilities in the disc gel. All the seven S-component appear to be dimor having molcular weight between 98000-94000. Any one of the seven f-component has a molecular size smaller than the S-component. Their molecular weight were estimated to be between 66800-50600.en
dc.description.provenanceMade available in DSpace on 2021-07-01T08:13:29Z (GMT). No. of bitstreams: 0
Previous issue date: 1983
en
dc.description.tableofcontents一、中文摘要 ……………………………………………………………………………………………………l
二、緒言 …………………………………………………………………………………………………………2
三、實驗材料 ……………………………………………………………………………………………………5
四、實驗方法 ……………………………………………………………………………………………………6
1. N6-(6-aminohexyl) adenosine 2,5 biphosphate 之合成……………………………………………6
2. Affinity gel 製備 ………………………………………………………………………………………7
3. G6PD 活性的測定 …………………………………………………………………………………………8
4.粗質 G6PD 之製備…………………………………………………………………………………………8
5. 粗質 G6PD 之純化…………………………………………………………………………………………9
6. 以電泳法鑑別 G6PD 酵素…………………………………………………………………………………10
7.以不同濃度 urea 處理 G6PD ……………………………………………………………………………11
8. 酵素的等電荷焦點電泳實驗………………………………………………………………………………12
9. Second Dimension Disc Electrophoresis 分析………………………………………………………12
五、結果與討論 …………………………………………………………………………………………………14
1. N6-(6-aminohexyl) adenosine 2,5 biphosphate 的合成及 Affinity gel 的製備………………14
2. G6PD 的純化 ………………………………………………………………………………………………15
3. G6PD 進行等電荷焦點電泳法實驗 ………………………………………………………………………16
4. Second Dimension Disc Electrophoresis 實驗………………………………………………………18
六、展 望 ………………………………………………………………………………………………………20
七、英文摘要 ……………………………………………………………………………………………………21
八、圖 表 ………………………………………………………………………………………………………27
九、誌 謝 ………………………………………………………………………………………………………44
十、參考文獻 ……………………………………………………………………………………………………45
dc.language.isozh-TW
dc.title人體紅血球葡萄醣六磷酸去氫的純化及性質的研究zh_TW
dc.titlePURIFICATION AND CHARACTERIZATION FOR GLUCOSE-6-PHOSPHATE DEHYDROGENASE FROM HUMAN ERYTHROCYTESen
dc.date.schoolyear71-2
dc.description.degree碩士
dc.relation.page52
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
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