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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 繆希椿(Shi-Chuen Miaw) | |
| dc.contributor.author | Chia-Kai Wu | en |
| dc.contributor.author | 吳茄愷 | zh_TW |
| dc.date.accessioned | 2021-06-15T04:52:09Z | - |
| dc.date.available | 2016-10-05 | |
| dc.date.copyright | 2011-10-05 | |
| dc.date.issued | 2011 | |
| dc.date.submitted | 2011-08-18 | |
| dc.identifier.citation | 1. Kataoka, K., M. Nishizawa and S. Kawai, Structure-function analysis of the maf oncogene product, a member of the bZip family. J Virol, 1993. 67: p. 2133-2141
2. Motohashi, H., J.A. Shavit, K. Igaeashi,M. Yamamoto, and J.D.Engel, The world according to Maf.pdf. Nucleic Acids Res, 1997. 25: p. 2953-2959. 3. Blank, V., Small Maf Proteins in Mammalian Gene Control: Mere Dimerization Partners or Dynamic Transcriptional Regulators? Journal of Molecular Biology, 2008. 376(4): p. 913-925. 4. Fujiwara KT., K.K., Nishizawa M., Two new members of the maf oncogene family, mafK and mafF, encode nuclear b-Zip proteins lacking putative trans-activator domain. Oncogene, 1993. 8: p. 2371-2380 5. Kotkow, K.J., and S.H. Orkin, Dependence of globin gene expression in mouse erythroleukemia cells on the NF-E2 heterodimer. Mol Cell Biol, 1995. 15: p. 4640-4647 6. Human mafG is a functional partner for p45 NF-E2 in activating globin gene expression. Blood. 89: p. 3925-3935 7. I-Cheng Ho, M.R.H., John W. Rooney and a.L.H. Glimcher, The Proto-Oncogene c-maf Is Responsible for Tissue-Specific Expression of IL-4. Cell, 1996. 85: p. 975-983. 8. Zhang, C., et al., MafA Is a Key Regulator of Glucose-Stimulated Insulin Secretion. Molecular and Cellular Biology, 2005. 25(12): p. 4969-4976. 9. Moriguchi, T., et al., MafB Is Essential for Renal Development and F4/80 Expression in Macrophages. Molecular and Cellular Biology, 2006. 26(15): p. 5715-5727. 10. Mears, A.J., et al., Nrl is required for rod photoreceptor development.Nature Genetics, 2001. 29(4): p. 447-452. 11. Grigoryan, G., A.W. Reinke, and A.E. Keating, Design of protein-interaction specificity gives selective bZIP-binding peptides. Nature, 2009. 458(7240): p. 859-864. 12. Yang Y., a.A.C., Large Maf Transcription factors; Cousins of AP-1 Proteins and Important Regulators of Cellular Differentiation. Einstein J Biol Med, 2007. 23: p. 2-11. 13. Motohashia, H., O'Connora. T., Katsuokab, F., Engelb, J.D. and Yamamoto, M., Integration and diversity of the regulatory network composed of Maf and CNC families of transcription factors. Gene, 2002. 294: p. 1-12. 14. James I. Kim., e., al., Requirement for the c-Maf transcription factor in crystallin gene regulation and lens development.pdf. Proc. Natl. Acad. Sci, 1999. 96: p. 3781-3785 15. Brian Z. Ring., S.P.C., Paul A. Overbeek., and Gregory S. Barsh., Regulation of mouse lens fiber cell development and differentiation by the Maf gene. Development, 2000. 127: p. 307-317. 16. Rajaram, N. and T.K. Kerppola, Synergistic Transcription Activation by Maf and Sox and Their Subnuclear Localization Are Disrupted by a Mutation in Maf That Causes Cataract. Molecular and Cellular Biology, 2004. 24(13): p. 5694-5709. 17. Kataoka, k., et al., Differentially expressed Maf family transcription factors, c-Maf and MafA, activate glucagon and insulin gene expression in pancreatic islet alpha- and beta-cells. J Mol Endocrinol, 2004. 32: p. 9-20. 18. Zhu, J., H. Yamane, and W.E. Paul, Differentiation of Effector CD4 T Cell Populations*. Annual Review of Immunology, 2010. 28(1): p. 445-489. 19. Murphy, K.M. and S.L. Reiner, Decision making in the immune system: The lineage decisions of helper T cells. Nature Reviews Immunology, 2002. 2(12): p. 933-944. 20. Tanaka, S., et al., The enhancer HS2 critically regulates GATA-3-mediated Il4 transcription in TH2 cells. Nature Immunology, 2010. 12(1): p. 77-85. 21. Romani, L., et al., Defective tryptophan catabolism underlies inflammation in mouse chronic granulomatous disease. Nature, 2008. 451(7175): p. 211-215. 22. Korn, T., et al., IL-17 and Th17 Cells. Annual Review of Immunology, 2009. 27(1): p. 485-517. 23. Patricia J. Dubin, J.K.K., IL-23 mediates inflammatory responses to mucoid Pseudomonas aeruginosa lung infection in mice. Am J Physiol Lung Cell Mol Physiol 2006. 292: p. L519-L528. 24. Nurieva, R.I., et al., Bcl6 Mediates the Development of T Follicular Helper Cells. Science, 2009. 325(5943): p. 1001-1005. 25. Yu, D., et al., Lineage specification and heterogeneity of T follicular helper cells. Current Opinion in Immunology, 2009. 21(6): p. 619-625. 26. James I. Kim, I.-C.H. and a.L.H.G. Michael J. Grusby, The Transcription Factor c-Maf Controls the Production of IL-4 but Not other Th2 cytokines.pdf. Immunity, 1999. 10(745-751). 27. Bauquet, A.T., et al., The costimulatory molecule ICOS regulates the expression of c-Maf and IL-21 in the development of follicular T helper cells and TH-17 cells. Nature Immunology, 2008. 10(2): p. 167-175. 28. Apetoh, L., et al., The aryl hydrocarbon receptor interacts with c-Maf to promote the differentiation of type 1 regulatory T cells induced by IL-27. Nature Immunology, 2010. 11(9): p. 854-861. 29. Nagy, A., Cre Recombinase: The Universal Reagent for Genome Tailoring. genesis, 2000. 26: p. 99-109 30. BrianI Sauer and N. Henderson., Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. Proc. Natl. Acad. Sci, 1988. 85: p. 5166-5170. 31. Hayashi, S., Efficient Recombination in Diverse Tissues by a Tamoxifen-Inducible Form of Cre: A Tool for Temporally Regulated Gene Activation/Inactivation in the Mouse. Developmental Biology, 2002. 244(2): p. 305-318. 32. Neal G. Copeland., Nancy A. Jenkins., and and Donald L. Court, RECOMBINEERING A POWERFUL NEW TOOL FOR MOUSE FUNCTIONAL GENOMICS. Nat. Rev. Genet., 2001. 2: p. 769-779 33. Liu, P., A Highly Efficient Recombineering-Based Method for Generating Conditional Knockout Mutations. Genome Research, 2003. 13(3): p. 476-484. 34. R., P.A., What makes the bacteriophage λ Red system useful for genetic engineering: Molecular Mechanism and biological function. FEMS Microbiol Lett., 2001. 201: p.9-14. 35. Lee, E., A Highly Efficient Escherichia coli-Based Chromosome Engineering System Adapted for Recombinogenic Targeting and Subcloning of BAC DNA. Genomics, 2001. 73(1): p. 56-65. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46042 | - |
| dc.description.abstract | 轉錄因子c-Maf調控第二型輔助T淋巴細胞(TH2)的IL-4、第十七型輔助T淋巴細胞(TH17)的IL-21以及第一型調節T細胞(Tr1)的IL-10表現。這些細胞激素對於個別細胞族群除了有重要的功能外，同時也是維持該細胞族群不可或缺的因子。由於c-Maf剔除小鼠於胚胎時期或是出生後幾天內死亡，因此沒有一個好的c-maf基因缺失的動物模式來探討c-Maf缺失對於體內免疫功能的影響。至目前為止，將c-Maf剔除小鼠的fetal liver轉殖進入RAG-1剔除小鼠，是現今可以得到c-Maf剔除T淋巴細胞的方法，可是無法持續繁殖此類實驗鼠。因此，使用條件性基因剔除小鼠是一個解決此問題的最好策略。在本篇論文中，利用DNA recombineering方式建構c-maf條件性剔除構成物。將進一步以此構成物來建立c-Maf條件性剔除小鼠，這將是一個相當有價值的小鼠模式來研究c-Maf在免疫功能中所扮演的角色。 | zh_TW |
| dc.description.abstract | The transcription factor c-Maf is an important transactivator for the IL-4, IL-21 and IL-10 production in TH2, TH17 and Tr1 (Type 1 regulatory T cells), respectively. These cytokines are important for the function of their cell subtypes. Convention c-Maf-deficient mice die within few days after birth. Therefore, there is obstacle to use c-Maf-deficient mice for studying c-Maf regulating immune fuction in vivo. Until now, the only way to investigate the effect of c-Maf deficiency in T cells was transferring the fetal liver of c-Maf null mice into RAG-1 knockout mice. However, these mice can not be propagated to maintain mice strain. The best strategy to study the role of c-Maf in T cells is to generate c-Maf conditional knockout mice. In these study, we built a c-Maf conditional knockout construct by DNA recombineering for generating c-Maf conditional knockout mice. It could be a valuable murine model for the studies of c-Maf mediated immune function. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T04:52:09Z (GMT). No. of bitstreams: 1 ntu-100-R98449011-1.pdf: 1080562 bytes, checksum: 551a57deaef348c4323de683b60d2245 (MD5) Previous issue date: 2011 | en |
| dc.description.tableofcontents | 目錄:
致謝 Ⅰ 中文摘要 Ⅱ Abstract Ⅲ 目錄 Ⅳ 1.背景介紹 1 (1)轉錄因子c-Maf 1 (2)輔助型CD4+ T淋巴細胞 2 (3)轉錄因子c-Maf在免疫系統中的角色 2 (4)c-Maf條件性剔除小鼠的重要性 3 2.實驗材料 6 2.(1) 化學藥品及試劑 6 2.(2) 酵素及抗體 7 2.(3) 實驗Kits 9 2.(4) 設備 9 2.(5) 培養液及緩衝液 10 3.方法 12 3.(1)勝任細胞的製備 a.氯化鈣(CaCl2)法製備 12 b.電穿孔法(Electroporation)製備 12 3.(2)轉型作用(Transformation) 熱休克轉型法(Heat-shock transformation) 13 a.使用氯化鈣(CaCl2)法之勝任細胞 13 電穿孔作用(Electroporation) 13 b.使用電穿孔法(Electroporation)之勝任細胞 13 3.(3)DNA重組工程(DNA Recombineering8 a. Gap repair (Retrieving) 13 b. Targeting 14 3.(4)檢驗大腸桿菌內質體DNA剔除作用 a.運用表現Cre recombinase 之EL350進行DNA剔除作用 14 b.運用表現Flp recombinase 之EL250進行DNA剔除作用 15 3.(5)建立c-Maf條件性剔除構成物(Conditional knockout construct: CKO) 16 3.(6)西方墨點法(Western blot) 17 3.(7)細胞培養和轉染(Transfection) 17 3.(8)蛋白質穩定性 18 3.(9) c-Maf變異型反轉錄病毒質體 18 4.結果 20 c-Maf條件性剔除構成物(CKO)的產生及DNA targeting 20 Wt和Y3F c-Maf蛋白質穩定性 21 c-Maf變異型反轉錄病毒質體 22 5.討論 23 c-Maf條件性剔除構成物(CKO)的產生及c-Maf條件性剔除小鼠之運用 23 Y3F c-Maf蛋白質穩定性與Wt c-Maf相似 25 c-Maf變異型反轉錄病毒質體 26 6.參考文獻 27 7.表 32 (1)使用Recombineering方式建構c-Maf條件性剔除構成物的引子序列 32 (2)延伸c-Maf條件性剔除構成物5′端序列所使的引子序列 33 (3)製造南方墨點法的探針所使用的引子序列: 33 8.附圖 34 圖一. DNA recombineering: Gap repair (Retrieving) 34 圖二. DNA recombineering: Targeting 36 圖三. 運用表現Cre recombinase 之EL350勝任細胞或是表現Flpe recombinase之EL250勝任細胞進行DNA剔除作用: 38 圖四. CKO p1+3+4 pSK+ 載體 40 圖五. CKO p1+3+4 pL253 載體 42 圖六. pL253FL (FL: Full-Length sequence of CKO) 44 圖七. pL253FL with Neomycin loxP cassette (pL253FLNeo) 46 圖八. pL253FC( pL253 Final Construct) 48 圖九. 使用Cre-loxP system檢視pL253FC質體是否具有基因剔除能力 50 圖十. 使用Flpe-FRT system檢驗pL253FC質體是否具有基因剔除能力 52 圖十一. 使用Flpe-FRT system和Cre-loxP system雙重檢驗pL253FC質體是否具有基因剔除能力 54 圖十二. pL253FCEP(pL253FC with extended part ) 56 圖十三. c-Maf CKO DNA targeting 58 圖十四.Wt和Y3F c-Maf蛋白質穩定性 60 圖十五. c-Maf變異型反轉錄病毒質體 62 | |
| dc.language.iso | zh-TW | |
| dc.subject | 條件性剔除構成物 | zh_TW |
| dc.subject | 條件性剔除小鼠 | zh_TW |
| dc.subject | DNA recombineering | en |
| dc.subject | c-Maf | en |
| dc.title | 建立c-maf條件性剔除構成物來產生c-Maf剔除小鼠 | zh_TW |
| dc.title | To build c-maf conditional knockout construct for generating c-Maf knockout mice | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 99-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 伍安怡(Betty-A Wu-Hsieh),司徒惠康 | |
| dc.subject.keyword | 條件性剔除小鼠,條件性剔除構成物, | zh_TW |
| dc.subject.keyword | c-Maf,DNA recombineering, | en |
| dc.relation.page | 63 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2011-08-18 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 免疫學研究所 | zh_TW |
| 顯示於系所單位： | 免疫學研究所 | |
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