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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 黃火鍊 | |
dc.contributor.author | Liang-Tai Wei | en |
dc.contributor.author | 魏良泰 | zh_TW |
dc.date.accessioned | 2021-06-13T03:14:42Z | - |
dc.date.available | 2007-08-01 | |
dc.date.copyright | 2006-08-01 | |
dc.date.issued | 2006 | |
dc.date.submitted | 2006-08-01 | |
dc.identifier.citation | 伍、參考文獻
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The doublesex proteins of Drosophila melanogaster bind directly to a sex-specific yolk protein gene enhancer. Embo J 10, 2577-82. Chase, B. A. and Baker, B. S. (1995). A genetic analysis of intersex, a gene regulating sexual differentiation in Drosophila melanogaster females. Genetics 139, 1649-61. Cline, T. W. and Meyer, B. J. (1996). Vive la difference: males vs females in flies vs worms. Annu Rev Genet 30, 637-702. Conaway, J. W., Florens, L., Sato, S., Tomomori-Sato, C., Parmely, T. J., Yao, T., Swanson, S. K., Banks, C. A., Washburn, M. P. and Conaway, R. C. (2005). The mammalian Mediator complex. FEBS Letters 579, 904-908. Coschigano, K. T. and Wensink, P. C. (1993). Sex-specific transcriptional regulation by the male and female doublesex proteins of Drosophila. Genes Dev 7, 42-54. Dynan, W. S. and Tjian, R. (1983). The promoter-specific transcription factor Sp1 binds to upstream sequences in the SV40 early promoter. Cell 35, 79-87. Dynlacht, B. D., Hoey, T. and Tjian, R. (1991). Isolation of coactivators associated with the TATA-binding protein that mediate transcriptional activation Cell 66, 563-576. Erdman, S. E. and Burtis, K. C. (1993). The Drosophila doublesex proteins share a novel zinc finger related DNA binding domain. Embo J 12, 527-35. Erdman, S. E., Chen, H. J. and Burtis, K. C. (1996). Functional and genetic characterization of the oligomerization and DNA binding properties of the Drosophila doublesex proteins. Genetics 144, 1639-52. Garrett-Engele, C. M., Siegal, M. L., Manoli, D. S., Williams, B. C., Li, H. and Baker, B. S. (2002). intersex, a gene required for female sexual development in Drosophila, is expressed in both sexes and functions together with doublesex to regulate terminal differentiation. Development 129, 4661-75. Hays, T. S., Deuring, R., Robertson, B., Prout, M. and Fuller, M. T. (1989). Interacting proteins identified by genetic interactions: a missense mutation in alpha-tubulin fails to complement alleles of the testis-specific beta-tubulin gene of Drosophila melanogaster. Mol Cell Biol 9, 875-84. Kim, Y.-J., Bjorklund, S., Li, Y., Sayre, M. H. and Kornberg, R. D. (1994). A multiprotein mediator of transcriptional activation and its interaction with the C-terminal repeat domain of RNA polymerase II. Cell 77, 599-608. Lee, T. I. and Young, R. A. (2000). Transcription of eukaryotic protein-coding genes. Annual Review of Genetics 34, 77-137. Lemon., B. and Tjian., R. (2000). Orchestrated response: a symphony of transcription factors for gene control Genes Dev. 14, 2551-2569. Li, H. and Baker, B. S. (1998a). Her, a gene required for sexual differentiation in Drosophila, encodes a zinc finger protein with characteristics of ZFY-like proteins and is expressed independently of the sex determination hierarchy. Development 125, 225-35. Li, H. and Baker, B. S. (1998b). hermaphrodite and doublesex function both dependently and independently to control various aspects of sexual differentiation in Drosophila. Development 125, 2641-51. Marin, I. and Baker, B. S. (1998). The evolutionary dynamics of sex determination. Science 281, 1990-4. Meisterernst, M., Roy, A. L., Lieu, H. M. and Roeder, R. G. (1991). Activation of class II gene transcription by regulatory factors is potentiated by a novel activity. Cell 66, 981-993 Myer, V. E. and Young, R. A. (1998). RNA polymerase II holoenzymes and subcomplexes. J Biol Chem 273, 27757-27760. Myers, L. C., Gustafsson, C. M., Bushnell, D. A., Lui, M., Erdjument-Bromage, H., Tempst, P. and Kornberg, R. D. (1998). The Med proteins of yeast and their function through the RNA polymerase II carboxy-terminal domain. Genes Dev. 12, 45-54. Myers, L. C. and Kornberg, R. D. (2000). Mediator of transcriptional regulation. Annual Review of Biochemistry 69, 729-749. Pinto, M. and Lobe, C. G. (1996). Products of the grg (Groucho-related gene) family can dimerize through the amino-terminal Q domain. J Biol Chem 271, 33026-31. Pultz, M. A. and Baker, B. S. (1995). The dual role of hermaphrodite in the Drosophila sex determination regulatory hierarchy. Development 121, 99-111. Pultz, M. A., Carson, G. S. and Baker, B. S. (1994). A genetic analysis of hermaphrodite, a pleiotropic sex determination gene in Drosophila melanogaster. Genetics 136, 195-207. Rachez, C. and Freedman, L. P. (2001). Mediator complexes and transcription. Curr Opin Cell Biol 13, 274-80. Ryner, L. C. and Baker, B. S. (1991). Regulation of doublesex pre-mRNA processing occurs by 3'-splice site activation. Genes Dev 5, 2071-85. Saluja, D., Vassallo, M. F. and Tanese, N. (1998). Distinct subdomains of human TAFII130 are required for interactions with glutamine-rich transcriptional activators. Mol Cell Biol 18, 5734-43. Sato, S., Tomomori-Sato, C., Banks, C. A., Parmely, T. J., Sorokina, I., Brower, C. S., Conaway, R. C. and Conaway, J. W. (2003). A mammalian homolog of Drosophila melanogaster transcriptional coactivator intersex is a subunit of the mammalian Mediator complex. J Biol Chem 278, 49671-4. Sato, S., Tomomori-Sato, C., Parmely, T. J., Florens, L., Zybailov, B., Swanson, S. K., Banks, C. A., Jin, J., Cai, Y., Washburn, M. P. et al. (2004). A set of consensus mammalian mediator subunits identified by multidimensional protein identification technology. Mol Cell 14, 685-91. Seipel, K., Georgiev, O. and Schaffner, W. (1992). Different activation domains stimulate transcription from remote ('enhancer') and proximal ('promoter') positions. Embo J 11, 4961-8. Siegal, M. L. and Baker, B. S. (2005). Functional conservation and divergence of intersex, a gene required for female differentiation in Drosophila melanogaster. Dev Genes Evol 215, 1-12. Stearns, T. and Botstein, D. (1988). Unlinked noncomplementation: isolation of new conditional-lethal mutations in each of the tubulin genes of Saccharomyces cerevisiae. Genetics 119, 249-60. Tanese, N. and Tjian, R. (1993). Coactivators and TAFs: a new class of eukaryotic transcription factors that connect activators to the basal machinery. Cold Spring Harb Symp Quant Biol 58, 179-85. Thompson, C. M., Koleske, A. J., Chao, D. M. and Young, R. A. (1993). A multisubunit complex associated with the RNA polymerase II CTD and TATA-binding protein in yeast. Cell 73, 1361-1375. Van Doren, M., Ellis, H. M. and Posakony, J. W. (1991). The Drosophila extramacrochaetae protein antagonizes sequence-specific DNA binding by daughterless/achaete-scute protein complexes. Develoment 113, 245-255. Verrijzer, C. P. and Tjian, R. (1996). TAFs mediate transcriptional activation and promoter selectivity. Trends in Biochemical Sciences 21, 338-342 Wang, Y., Li, Y., Zeng, W., Zhu, C., Xiao, J., Yuan, W., Wang, Y., Cai, Z., Zhou, J., Liu, M. et al. (2004). IXL, a new subunit of the mammalian Mediator complex, functions as a transcriptional suppressor. Biochem Biophys Res Commun 325, 1330-8. Waterbury, J. A., Jackson, L. L. and Schedl, P. (1999). Analysis of the doublesex female protein in Drosophila melanogaster: role on sexual differentiation and behavior and dependence on intersex. Genetics 152, 1653-67. Younger-Shepherd, S., Vaessin, H., Bier, E., Jan, L. Y. and Jan, Y. N. (1992). deadpan, an essential pan-neural gene encoding an HLH protein, acts as a denominator in Drosophila sex determination. Cell 70, 911-22. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/31553 | - |
dc.description.abstract | 中文摘要
在過去的研究中,對Drosophila melanogaster性別決定機制已瞭解得很清楚,其中一個名為intersex (ix)的基因,在雌性個體會與doublesex (dsx)共同作用於此調節層級的最底層。Drosophila的ix在雌雄個體上皆會表現,但對於體細胞性別發展只作用於雌性,在雄性個體上並無影響。其胺基酸序列並沒有可辨識的DNA結合區能直接與DNA結合,但是有類似轉錄活化結構功能區,可與其他蛋白質產生交互作用,與DSXF共同作用時,可能扮演了相當於DSXM C端的功能。 Mediator complex發現於酵母菌,為由20個以上的分子所構成的蛋白複合體,對轉錄的調控扮演著相當重要的角色,其中Brower等人(2002)與Wang等人(2004)皆選殖到一個為哺乳類Mediator complex次分子的新基因,是Drosophila ix的同源體,北方墨點中驗證了在許多組織中皆有表現,且蛋白主要分佈於細胞核內,推測其功能為轉錄調節者。 本實驗室同樣也在鯉魚的睪丸中發現了類似Drosophila intersex的基因(cIX),鑒於intersex在果蠅中所扮演性別決定的重要功能,對於此基因在鯉魚中所擔任的功能而感到興趣,為了進一步探討cIX的功能,本論文完成cIX全長cDNA,設計並建構cIX的重組DNA,再利用純化之重組蛋白誘導抗體。根據PCR以及西方墨點的實驗結果,顯示cIX是一個廣泛存在於各個鯉魚組織的蛋白質,並沒有組織特異性。然而也發現cIX有主要兩種不同的形式存在,另外在與其他物種的同源蛋白質的胺基酸序列比對中,顯示此類蛋白質在C端的保守性很高,N端的變異性則較大,至於這些資訊所代表的意義則需要再進一步的研究。 | zh_TW |
dc.description.abstract | Abstract
The sex-determination of Drosophila melanogaster has been known clearly in previous studies. There is a gene called intersex (ix), which functions together with doublesex (dsx) at the bottom of the hierarchy in the sex deternination females. Drosophila ix is expressed in both sex, but only functions in female somatic cell. It has no effect on male. There is no recognizable DNA-binding domain on the amino acid sequence of ix for binding to DNA, but it contains a region like transcriptional activation domains to interact with other proteins, and ix might play a similar role as the C-terminal of DSXM when acts together with DSXF. Mediator complex was found in yeast, it is a protein complex composed over 20 subunits, and plays an essential role in the regulation of transcription. Brower et al. (2002) and Wang et al. (2004) also have cloned a navel gene of mammalian Mediator complex subunit, which is the homolog of Drosophila ix. Northern blot analysis demonstrated that ix is expressed in various tissues, and the protein is localized mainly to nucleus, supporting its function as a transcriptional regulator. Our laboratory also found a gene like Drosophila intersex in the testis of Cyprinus carpio (cIX). According to the significant function played by Drosophila ix in sex-determination, we are interested in the function of this gene in carp. To study the functions of cIX further, I cloned the full-length cDNA of cIX. The recombinant protein is produced and purified to serve as the antigen to induce antibody. The results of PCR and western blot analysis demonstrated that cIX is broadly existing in various tissue of carp, when the tissue specificity is not distinct. I also found that cIX has two mainly form, and the alignment of the homologs of other species indicates that IX proteins are conserved in C-terminal, and shows variation in N-terminal. The meaning of these information needs further study. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T03:14:42Z (GMT). No. of bitstreams: 1 ntu-95-R92b43012-1.pdf: 1725534 bytes, checksum: eb168f5d4973cba3b98b720943a976c2 (MD5) Previous issue date: 2006 | en |
dc.description.tableofcontents | 目錄
目錄 I 中文摘要 III Abstract IV 壹、緒論 1 (一) Drosophila melanogaster性別決定機制 1 (二) Drosophila melanogaster之Intersex 2 (三) Mediator Complex 4 (四) Intersex of the Mammalian Mediator Complex 5 (五) Intersex of Cyprinus carpio 6 貳、材料與方法 7 (一) 實驗動物 7 (二) 鯉魚intersex(cIX)重組蛋白之製備 7 (三) Intersex重組蛋白抗體之誘導與純化 14 (四) 西方墨點法(Western blot) 16 (五) 免疫沉澱法(immunoprecipitation) 18 (六) 二維電泳 19 (七) 鯉魚intersex(cIX)全長mRNA之定序 22 (八) 各物種Intersex蛋白質序列分析 22 叄、結果 24 (一) Cyprinus carpio intersex基因之選殖 24 (二) C.carpio 與D.rerio之intersex序列比較 25 (三) Cyprinus carpio intersex預測之胺基酸序列的比較 25 (四) Cyprinus carpio intersex重組蛋白之製備 26 (五) Cyprinus carpio intersex抗體之製備 27 (六) 二維電泳分析 27 肆、討論 29 (一) Cyprinus carpio intersex基因之選殖 29 (二) Cyprinus carpio intersex之胺基酸序列的比較 30 (三) Cyprinus carpio intersex重組蛋白抗體 31 (四) 睪丸萃取液經免疫沈澱後的二維電泳分析 31 (五) 未來可能的研究方向 32 伍、參考文獻 34 陸、圖表 37 圖1- Drosophila melanogaster的性別決定調節層級 38 圖2- Drosophila melanogaster之Doublesex(DSX)與Intersex(IX)示意圖 39 圖3- Mediator Complex於轉錄起始時訊息的傳遞示意圖 39 圖4- 鯉魚組織cDNA基因庫進行cIX之PCR結果 40 圖5- C.carpio Intersex mRNA與胺基酸序列(normal form) 41 圖6- C.carpio Intersex mRNA與胺基酸序列(short form) 42 圖7- IX-normal、IX-short與斑馬魚IX之序列比較 43 圖8- 各物種之intersex胺基酸序列比對與UPMGA系統發生樹 45 圖9-. cIX重組蛋白之表現質體IX-pET-1 MAP 47 圖10- IPTG誘導表現之結果 48 圖11- 兔子抗cIX血清之測試 49 圖12- cIX蛋白於鯉魚各組織之表現 50 圖13- 鯉魚睪丸組織進行免疫沈澱後洗滌之劃分 51 圖14- 鯉魚睪丸組織免疫沈澱之二維電泳分析 52 表1- 引子(primer)列表 53 | |
dc.language.iso | zh-TW | |
dc.title | 鯉魚Intersex基因之選殖與定性之研究 | zh_TW |
dc.title | Cloning and Characterization of Carp Intersex Gene | en |
dc.type | Thesis | |
dc.date.schoolyear | 94-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 柯逢春,李明亭 | |
dc.subject.keyword | 鯉魚, | zh_TW |
dc.subject.keyword | intersex,carp,mediator complex, | en |
dc.relation.page | 53 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2006-08-01 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
顯示於系所單位: | 分子與細胞生物學研究所 |
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