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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.author | Ben-Nen Wang | en |
| dc.contributor.author | 王本甯 | zh_TW |
| dc.date.accessioned | 2021-07-01T08:20:39Z | - |
| dc.date.available | 2021-07-01T08:20:39Z | - |
| dc.date.issued | 1998 | |
| dc.identifier.citation | 陳金龍（1990）國立台灣大學生化科學研究所碩士論文 蔡偉博（1991）國立台灣大學生化科學研究所碩士論文 黃彥華（1993）國立台灣大學生化科學研究所碩士論文 羅清維（1994）國立台灣大學生化科學研究所碩士論文 賴明龍（1993）國立台灣大學生化科學研究所博士論文 餘榮熾（1993）國立台灣大學生化科學研究所博士論文 楊允馨（1994）國立台灣大學生化科學研究所碩士論文 蕭雅鈴（1996）國立台灣大學生化科學研究所碩士論文 黃彥華（1997）國立台灣大學生化科學研究所博士論文 Alder A. J., Scheller A., Hpffman, Y., and Robins D. M. (1991) Mol. Endocrinol. 5, 1587-1596 Aris, J. P. and Blobel, G. (1991) Methods Enzymol. 194, 735-749 Beato, M. (1989) Cell 56, 335-344 Bradshaw, B. S. and Wolfe, H. G. (1977) Biol. Reprod. 16, 292-297 Brinkmann, A. O., Kuiper, G. G., Ris-Stalpers, C., van Rooij, H. C., Romalo, G., Trifiro, M., Mulder E., Pinsky, L., Schweikert, H. U. and Trapman, J. (1991) J. Steroid Biochem. Mol. Biol. 40 (1-3), 349-352, 1991 Cato, A. C. B., Skroch, P. S. Mary, P. T., Snoek, P. and Ponta, H. (1988) EMBO J. 7, 1403-1410 Celis, L., Claessens, F., Peter, B., Heyens, W., Verhoeven, G., Rombauts, W. (1993) Mol. Cell. Endocrinal. 94, 165-172 Chang, C., Kokontis, J. and Liao, S. (1988) Science 240, 324-326 Chung, C. T>, Niemela, S. L. and Miller, R. H. (1989) Proc. Natl. Acad. Sci. USA 86, 2172-2175 Classens, F., Celid, L., De Vos, P., Peeters, B., Heyns, W., Verhoeven, G. and Rombauts, W. (1993) Biochem. Biophys. Res. Commun. 191(2), 688-694 Cleutjiens, K. B., Eekenlen, C. C., Korput, H. A., Brinkmann, A. O. and Trapman, J. (1996) J. Biol. Chem. 271(11), 6379-6388 Cleutjiens, K. B., Korput, H. A., Eekelen, C. C., Rooij, H. C., Faber, P. W. and Trapman, J. (1997) Mol. Endocrinol. 11(2), 148-161 Dijck, P. V., Winderickx., J., Heyns, W. and Verhoeven, G. (1989) Mol. Cell. Endocrinol. 64, 195-204 Fabre, S. Manin, M., Paihoux, E., Veyssiere, G., Jean C. (1994) J. Biol. Chem. 169, 5857-5864 Ham, J., Thomoson, A., Needham, N. Webb, P. and Parker, M., (1988) Nucleic Acids Res. 16, 5263-5276 Ho, K. C., Marschke, K. B., Tan, J. A., Power, S. G. A., Wilson, E. M. and French, F. S. (1993) J. Biol. Chem. 168, 27226-27235 Huang, Y-H., Luo, C-W., Yu, L-C., Chu, S-T., Chen, Y-H. (1995) Biophysical J. 69, 2084-2089 Lucas, P. C. and Granner, D. K. (1992) Annu. Rev. Biochem. 61, 1131-1173 Kemppainen, J. A., Lane, M. V., Sar, M. and Wilson, E. M. (1992) J. Biol. Chem. 267(2), 968-974 Mitchell, P. J., and Tjian, R. (1989) Science 245, 371-378 Murtha, P., Tindall, D. J. and Young, C. Y. F. (1993) Biochem. 32, 6459-6464 Orti, E., Bodwell, J. E., and Munck, A. (1992) Endocrine Rev. 13, 105-128 Pang, S. F., Chow, P. H. and Wong, T. M. (1979) J. Reprod Fertil. 56, 129-132 Peitz, B. and Olds-Clarke, P. (1986) Biol. Reprod. 35, 608-617 Price, D. and Williams-Ashman, H. G. (1961) in Sex and Iternal Secretions (Young, W. C., ed.), vol, 1, pp. 366-448, Williams & Wilkins, Baltimore Qween, K., Dhabuwala, C. B. and Prerrpoint, C. G. (1981)J. Reprod Fertil. 62, 423-426 Rennie, P. S>, Bruchovsky, N., Leco, K. T., Sheppard, P. C., McQueen, S. A., Cheng, H., Snoek, P., Hamel, A., Bock, M. E. (1993) Mol. Endocrinol. 7, 23-36 Riegman, P. H. J., Vlietstra, R. J., van der Korput, J. A. G. M., Brinkmann, A. O. and Trapman, J. (1991) Mol. Endocrinol. 5, 1921-1930 Rushmere, N. K., Parker, M. G. and Davies, P. (1987) Mol. Endocrinol. 5, 1921-1930 Sanger, F., Nicklen, S. and Coulson, A. R. (1977) Proc. Natl. Acad. Sci. USA 74, 5463 Schuur, E. R., Henderson, G. A., Kmetec, L. A., Miller, J. D., Lamparski, H. G. and Henderson, D. R. (1996)J. Biol. Chem. 271(12), 7043-7051 Steitz, J. and Aumuller, G. (1991) Andrologia 24, 27-31 Simental, J. A., Sar, M., Lane, M. V., French, F. S. and Wilson, E. M. (1991) J. Biol. Chem. 266(1), 510-518 Strahle, U., Schmid, W., and Schutz, G. (1988) EMBOJ. 7, 3389-3395 Tan, J. A., Marschke, K. B., Ho, K.C., Perry S. T., Wilson, E. M. and French F, S. (1992) J. Biol. Chem. 267,4456-4466 Trus, M. and Beato, M. (1993) Endocrine Rev. 14, 459-479 Veldscholte J., Berrevoets C. A., Ris-Stalpers C., Kuiper G. G., Jenster G., Trapman J., Brinkmann A. O., and Mulder E. (1992) J. Steroid Biochem. & Mol. Bio. 41(3-8), 665-9 White, I. J. (1958) Anim. Breeding 26, 709-711 Yamamoto, K. R. (1985) Annu. Rev. Genet. 19, 209-252 Yu, L-C., Chen, J-L., Tasi, W-B. and Chen, Y-H. (1993) Biochem. J. 196, 571-576 Yu, L-C., Hsiao, Y-L., Yang, Y-H., Lin, M. and Chen, Y-H. (1997) Biochem. Biophys. Res. Commun. 231, 106-110 Zhou, Z., Lane, M. V., KeppINEN, j. a., French, F. S. and Wilson, E. M. (1995) Mol. Endocriol. 9(2), 208-218 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76358 | - |
| dc.description.abstract | 儲精囊等雄性附屬性腺分泌液占精液體積的二分之一以上，其中含有許多影響生殖力的因數。小白鼠儲精囊自體抗原（SVA）純化自小白鼠儲精囊分泌液，已知SVA可與精蟲結合並影響其活動力，且其基因表現受雄性素調控，故SVA在生殖功能上可能具有一定的重要性。 由比對分析的結果，在SVA基因序列上發現十個與雄性素反應單元（ARE）保守序列相似性大於50％的區域。我們建構了8個包含不同片段ARE-like elements的報導質體，以攝護腺癌細胞LNCaP作為表現系統，針對其中五個ARE-like elements，探討與雄性素所調控基因表現之間的關係。結果發現，當ARE-a, ARE-b, TATA-Box同時存在報導基因上游時，基因表現可受雄性素刺激達2.5倍；而當ARE-c, ARE-d, TATA-Box同時存在報導基因上游時，基因表現可受雄性素刺激達4.8倍。 另外，我們使用Electrophoresis Mobility Shift Assay (EMSA)分析雄性素受體與SVA?動子的結合區域，結果發現儲精囊細胞核萃取物與SVA基因上-250?＋8的區域有結合，且結合的程度隨著核萃取物濃度增加而增加。相較於重組的大鼠雄性素受體DNA結合區域（rAR(DBD)）與此段序列的結合，儲精囊核萃取物的結合程度明顯大了許多，因而猜測有某些儲精囊特有的因數與之結合。 本論文雖未能完整的解答雄性素與SVA基因表現之間的關係，所提供初步的研究結果可作為未來研究的參考。 | zh_TW |
| dc.description.abstract | Semen comes mainly from the secretions of seminal vesicle and other accessory glands. An autoantigen, the seminal vesicle autoantigen (SVA), has been purified from seminal vesicle secretion. It is a 19 kDa androgen-dependent glycoprotein. The preliminary data from our laboratory indicate that SVA is a sperm mobility inhibitor with the ability to suppress the BSA-induce sperm capacitation. By comparison, ten potential androgen response elements (AREs) have been mapped on the gene. In this work, we ligated different fragments of SVA promoter and intronic regions on the upstream of a reporter gene chloramphenicol acetyltransferase (CAT). Eight plasmids were constructed and their transcriptional activities were evaluated by CAT assay. Including ARE-a, ARE-b, and TATA-box together, the plasmid could enhance 2.5 folds of CAT expression under induced by 10 nM R1881. Under the same condition, ARE-c, ARE-d, and TATA-box were cooperated to enhance 4.8 folds of CAT expression. Our preliminary study suggested that the protein(s) from nuclear extract of seminal vesicle could bind to -250?+8 region of SVA gene. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-01T08:20:39Z (GMT). No. of bitstreams: 0 Previous issue date: 1998 | en |
| dc.description.tableofcontents | 目 錄……………………………………………………i 圖表目錄……………………………………………………iii 中文摘要……………………………………………………I ABSTRACT……………………………………………………II 縮寫表……………………………………………………III 第一章 緒論……………………………………………………1 1．1 儲精囊分泌液與生殖的相關性……………………………………………………1 1．2 小白鼠儲精囊自體抗原……………………………………………………2 1．2．1 特性及功能Characters of SVA……………………………………………………2 1．2．2 基因層次的研究Molecular Studies……………………………………………………2 1．3 雄性素與基因調控……………………………………………………3 1．3．1 雄性素Androgen……………………………………………………3 1．3．2 雄性素受體Androgen Receptor, AR……………………………………………………3 1．3．3 雄性素反應單元Androgen Response Element, ARE……………………………………………………4 1．3．4 可能作用機制Possible Mechanism of AR Action……………………………………………………5 1．4 本論文的研究方向……………………………………………………6 第二章 實驗設計與方法……………………………………………………15 2．1 實驗流程大綱……………………………………………………15 2．2 實驗設計……………………………………………………16 2．2．1 載體建構（圖2-1）Plasmids Construction……………………………………………………16 2．2．2 質體轉殖與基因表現Transfection and Expression……………………………………………………16 2．2．3 雄性素受器與DNA結合序列分析Interactions of AR and SVA Promoter Region……………………………………………………17 2．3 材料與方法MATERIAL AND METHODS……………………………………………………18 2．3．1 細胞培養與質體轉殖Cell Culture and Transfection……………………………………………………18 2．3．1．1 一般培養Cell Culture……………………………………………………18 2．3．1．2 細胞生長速率之計算Population Doubling……………………………………………………19 2．3．1．3 質體轉殖Plasmids Transfection……………………………………………………19 2．3．2 建構含有SVA?動子序列以及報導基因的質體Construction of Plasmids Include SVA Gene and Reporter Gene……………………………………………………20 2．3．2．1 以聚合脢連鎖反應製備SVA?動子片段Polymerase Chain Reaction, PCR……………………………………………………20 2．3．2．2 DNA片段溶離Elution DNA Fragment from Agarose Gel……………………………………………………21 2．3．2．3 DNA鏈合Ligation……………………………………………………22 2．3．2．4 質體轉型Transformation……………………………………………………23 2．3．2．5 小量質體抽取Plasmids Purification……………………………………………………24 2．3．2．6 大量質體抽取Plasmids Purification……………………………………………………25 2．3．2．7 洋菜膠體電泳Agarose Gel Electrophoresis……………………………………………………25 2．3．2．8 DNA定序DNA Sequencing……………………………………………………26 2．3．3 CAT Assay……………………………………………………29 2．3．4 細胞核萃取物之製備與分析方法Prepare and Analysis of Nuclear Extracts……………………………………………………31 2．3．4．1 細胞核萃取物之製備Prepare of Nuclear Extract……………………………………………………31 2．3．4．2 蛋白質定量Protein Concentration Definition……………………………………………………34 2．3．4．3 SDS-聚丙烯醯胺膠體電泳SDS-PAGE……………………………………………………35 2．3．4．4 Serva Blue R染色法Serva Blue R Staining……………………………………………………36 2．3．4．5 西方墨點法Western Blotting……………………………………………………36 2．3．6 Electrophoresis Mobility Shift Aaasy (EMSA)……………………………………………………39 第三章 實驗結果……………………………………………………47 3．1 質體建構PLASMIDS CONSTRUCTION……………………………………………………47 3．2 細胞培養CELL CULTURE……………………………………………………48 3．3 轉錄起始功能測定TRANSCRIPTIONAL ACTIVITY ASSAY……………………………………………………48 3．4 EMSA……………………………………………………49 第四章 討論……………………………………………………61 參考資料……………………………………………………644 附錄一……………………………………………………67 附錄二……………………………………………………68 | |
| dc.language.iso | zh-TW | |
| dc.title | 小白鼠儲精囊自體抗原啟動子之研究 | zh_TW |
| dc.title | Studies of Mouse Seminal Vesicle Autoantigen Promoter | en |
| dc.date.schoolyear | 86-2 | |
| dc.description.degree | 碩士 | |
| dc.relation.page | 68 | |
| 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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