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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 黃楓婷(Feng-Ting Huang) | |
dc.contributor.author | Wen-Chuang Hsieh | en |
dc.contributor.author | 謝文娟 | zh_TW |
dc.date.accessioned | 2021-06-08T00:05:47Z | - |
dc.date.copyright | 2013-08-27 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-08-13 | |
dc.identifier.citation | Aoufouchi, S., Faili, A., Zober, C., D'Orlando, O., Weller, S., Weill, J. C., & Reynaud, C. A. (2008). Proteasomal degradation restricts the nuclear lifespan of AID. J Exp Med, 205(6), 1357-1368.
Aravind, L., & Koonin, E. V. (2001). Prokaryotic homologs of the eukaryotic DNA-end-binding protein Ku, novel domains in the Ku protein and prediction of a prokaryotic double-strand break repair system. Genome Res, 11(8), 1365-1374. Auerbach, A. D. (2009). Fanconi anemia and its diagnosis. Mutat Res, 668(1-2), 4-10. Bachrati, C. Z., & Hickson, I. D. (2008). RecQ helicases: guardian angels of the DNA replication fork. Chromosoma, 117(3), 219-233. Barreto, V., Reina-San-Martin, B., Ramiro, A. R., McBride, K. M., & Nussenzweig, M. C. (2003). C-terminal deletion of AID uncouples class switch recombination from somatic hypermutation and gene conversion. Mol Cell, 12(2), 501-508. Bassing, C. H., Swat, W., & Alt, F. W. (2002). The mechanism and regulation of chromosomal V(D)J recombination. Cell, 109 Suppl, S45-55. Blaho, V. A., Buczynski, M. W., Dennis, E. A., & Brown, C. R. (2009). Cyclooxygenase-1 orchestrates germinal center formation and antibody class-switch via regulation of IL-17. J Immunol, 183(9), 5644-5653. Bohr, V. A. (2008). Rising from the RecQ-age: the role of human RecQ helicases in genome maintenance. Trends Biochem Sci, 33(12), 609-620. Bottaro, A., Lansford, R., Xu, L., Zhang, J., Rothman, P., & Alt, F. W. (1994). S region transcription per se promotes basal IgE class switch recombination but additional factors regulate the efficiency of the process. EMBO J, 13(3), 665-674. Bransteitter, R., Pham, P., Scharff, M. D., & Goodman, M. F. (2003). Activation-induced cytidine deaminase deaminates deoxycytidine on single-stranded DNA but requires the action of RNase. Proc Natl Acad Sci U S A, 100(7), 4102-4107. Casellas, R., Nussenzweig, A., Wuerffel, R., Pelanda, R., Reichlin, A., Suh, H., Qin, X. F., Besmer, E., Kenter, A., Rajewsky, K., Nussenzweig, M. C. (1998). Ku80 is required for immunoglobulin isotype switching. EMBO J, 17(8), 2404-2411. Chaudhuri, J., & Alt, F. W. (2004). Class-switch recombination: interplay of transcription, DNA deamination and DNA repair. Nat Rev Immunol, 4(7), 541-552. Chaudhuri, J., Basu, U., Zarrin, A., Yan, C., Franco, S., Perlot, T., Vuong, B., Wang, J., Phan, R. T., Datta, A., Manis, J., Alt, F. W. (2007). Evolution of the immunoglobulin heavy chain class switch recombination mechanism. Adv Immunol, 94, 157-214. Chaudhuri, J., Tian, M., Khuong, C., Chua, K., Pinaud, E., & Alt, F. W. (2003). Transcription-targeted DNA deamination by the AID antibody diversification enzyme. Nature, 422(6933), 726-730. Conticello, S. G., Thomas, C. J., Petersen-Mahrt, S. K., & Neuberger, M. S. (2005). Evolution of the AID/APOBEC family of polynucleotide (deoxy)cytidine deaminases. Mol Biol Evol, 22(2), 367-377. Coulon, S., Gaillard, P. H., Chahwan, C., McDonald, W. H., Yates, J. R., 3rd, & Russell, P. (2004). Slx1-Slx4 are subunits of a structure-specific endonuclease that maintains ribosomal DNA in fission yeast. Mol Biol Cell, 15(1), 71-80. Coulon, S., Noguchi, E., Noguchi, C., Du, L. L., Nakamura, T. M., & Russell, P. (2006). Rad22Rad52-dependent repair of ribosomal DNA repeats cleaved by Slx1-Slx4 endonuclease. Mol Biol Cell, 17(4), 2081-2090. Crossan, G. P., van der Weyden, L., Rosado, I. V., Langevin, F., Gaillard, P. H., McIntyre, R. E., Gaillard, P. H., McIntyre, R. E., Gallagher, F., Kettunen, M. I., Lewis, D. Y., Brindle, K., Arends, M. J., Adams, D. J., Patel, K. J. (2011). Disruption of mouse Slx4, a regulator of structure-specific nucleases, phenocopies Fanconi anemia. Nat Genet, 43(2), 147-152. Daniels, G. A., & Lieber, M. R. (1995). RNA:DNA complex formation upon transcription of immunoglobulin switch regions: implications for the mechanism and regulation of class switch recombination. Nucleic Acids Res, 23(24), 5006-5011. de Winter, Johan P., & Joenje, Hans. (2009). The genetic and molecular basis of Fanconi anemia. Mutat Res, 668(1–2), 11-19. Delpy, L., Le Bert, M., Cogne, M., & Khamlichi, A. A. (2003). Germ-line transcription occurs on both the functional and the non-functional alleles of immunoglobulin constant heavy chain genes. Eur J Immunol, 33(8), 2108-2113. Di Noia, J. M., & Neuberger, M. S. (2007). Molecular mechanisms of antibody somatic hypermutation. Annu Rev Biochem, 76, 1-22. Di Noia, J., & Neuberger, M. S. (2002). Altering the pathway of immunoglobulin hypermutation by inhibiting uracil-DNA glycosylase. Nature, 419(6902), 43-48. Dickerson, S. K., Market, E., Besmer, E., & Papavasiliou, F. N. (2003). AID mediates hypermutation by deaminating single stranded DNA. J Exp Med, 197(10), 1291-1296. Dudley, D. D., Chaudhuri, J., Bassing, C. H., & Alt, F. W. (2005). Mechanism and control of V(D)J recombination versus class switch recombination: similarities and differences. Adv Immunol, 86, 43-112. Durandy, A. (2003). Activation-induced cytidine deaminase: a dual role in class-switch recombination and somatic hypermutation. Eur J Immunol, 33(8), 2069-2073. Ehrenstein, Michael R., Rada, Cristina, Jones, Anne-Marie, Milstein, Cesar, & Neuberger, Michael S. (2001). Switch junction sequences in PMS2-deficient mice reveal a microhomology-mediated mechanism of Ig class switch recombination. Proc Natl Acad Sci U S A, 98(25), 14553-14558. Fekairi, S., Scaglione, S., Chahwan, C., Taylor, E. R., Tissier, A., Coulon, S., Dong, M. Q., Ruse, C., Yates, J. R., 3rd, Russell, P., Fuchs, R. P., McGowan, C. H., Gaillard, P. H. (2009). Human SLX4 is a Holliday junction resolvase subunit that binds multiple DNA repair/recombination endonucleases. Cell, 138(1), 78-89. Fricke, W. M., & Brill, S. J. (2003). Slx1-Slx4 is a second structure-specific endonuclease functionally redundant with Sgs1-Top3. Genes Dev, 17(14), 1768-1778. Harriman, G. R., Bradley, A., Das, S., Rogers-Fani, P., & Davis, A. C. (1996). IgA class switch in I alpha exon-deficient mice. Role of germline transcription in class switch recombination. J Clin Invest, 97(2), 477-485. Hoege, C., Pfander, B., Moldovan, G. L., Pyrowolakis, G., & Jentsch, S. (2002). RAD6-dependent DNA repair is linked to modification of PCNA by ubiquitin and SUMO. Nature, 419(6903), 135-141. Honjo, T., Kinoshita, K., & Muramatsu, M. (2002). Molecular mechanism of class switch recombination: linkage with somatic hypermutation. Annu Rev Immunol, 20, 165-196. Honjo, T., Nagaoka, H., Shinkura, R., & Muramatsu, M. (2005). AID to overcome the limitations of genomic information. Nat Immunol, 6(7), 655-661. Imai, K., Slupphaug, G., Lee, W. I., Revy, P., Nonoyama, S., Catalan, N., Yel, L., Forveille, M., Kavli, B., Krokan, H. E., Ochs, H. D., Fisher, A., Durandy, A. (2003). Human uracil-DNA glycosylase deficiency associated with profoundly impaired immunoglobulin class-switch recombination. Nat Immunol, 4(10), 1023-1028. Jacob, J., Przylepa, J., Miller, C., & Kelsoe, G. (1993). In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. III. The kinetics of V region mutation and selection in germinal center B cells. J Exp Med, 178(4), 1293-1307. Jung, D., & Alt, F. W. (2004). Unraveling V(D)J recombination; insights into gene regulation. Cell, 116(2), 299-311. Jung, S., Rajewsky, K., & Radbruch, A. (1993). Shutdown of class switch recombination by deletion of a switch region control element. Science, 259(5097), 984-987. Kaliraman, V., & Brill, S. J. (2002). Role of SGS1 and SLX4 in maintaining rDNA structure in Saccharomyces cerevisiae. Curr Genet, 41(6), 389-400. Kang, S. H., Kim, G. R., Seong, M., Baek, S. H., Seol, J. H., Bang, O. S., Ovaa, H., Tatsumi, K., Komatsu, M., Tanaka, K., Chung, C. H. (2007). Two novel ubiquitin-fold modifier 1 (Ufm1)-specific proteases, UfSP1 and UfSP2. J Biol Chem, 282(8), 5256-5262. Khamlichi, AA, Glaudet, F, Oruc, Z, Denis, V, Le Bert, M, & Cogne, M. (2004). Immunoglobulin class-switch recombination in mice devoid of any S mu tandem repeat. Blood, 103(10), 3828-3836. Kim, Y., Lach, F. P., Desetty, R., Hanenberg, H., Auerbach, A. D., & Smogorzewska, A. (2011). Mutations of the SLX4 gene in Fanconi anemia. Nat Genet, 43(2), 142-146. Koo, B. H., Le Goff, C., Jungers, K. A., Vasanji, A., O'Flaherty, J., Weyman, C. M., & Apte, S. S. (2007). ADAMTS-like 2 (ADAMTSL2) is a secreted glycoprotein that is widely expressed during mouse embryogenesis and is regulated during skeletal myogenesis. Matrix Biol, 26(6), 431-441. Kotnis, Ashwin, Du, Likun, Liu, Chonghai, Popov, Sergey W, & Pan-Hammarstrom, Qiang. (2009). Non-homologous end joining in class switch recombination: the beginning of the end. Philos T Roy Soc B, 364(1517), 653-665. Kracker, S., & Durandy, A. (2011). Insights into the B cell specific process of immunoglobulin class switch recombination. Immunol Lett, 138(2), 97-103. Le Goff, C., Morice-Picard, F., Dagoneau, N., Wang, L. W., Perrot, C., Crow, Y. J., Bauer. F., Flori, E., Prost-Squarcioni, C., Krakow, D., Ge, G., Greenspan, D. S., Bonnet, D., Le Merrer, M., Munnich, A., Apte, S. S., Cormier-Daire, V. (2008). ADAMTSL2 mutations in geleophysic dysplasia demonstrate a role for ADAMTS-like proteins in TGF-beta bioavailability regulation. Nat Genet, 40(9), 1119-1123. Lee, H., Trott, J. S., Haque, S., McCormick, S., Chiorazzi, N., & Mongini, P. K. (2010). A cyclooxygenase-2/prostaglandin E2 pathway augments activation-induced cytosine deaminase expression within replicating human B cells. J Immunol, 185(9), 5300-5314. Li, Z., Scherer, S. J., Ronai, D., Iglesias-Ussel, M. D., Peled, J. U., Bardwell, P. D., Zhuang, M., Lee, K., Martin, A., Edelmann, W., Scharff, M. D. (2004). Examination of Msh6- and Msh3-deficient mice in class switching reveals overlapping and distinct roles of MutS homologues in antibody diversification. J Exp Med, 200(1), 47-59. Lieber, M. R. (2010). The mechanism of double-strand DNA break repair by the nonhomologous DNA end-joining pathway. Annu Rev Biochem, 79, 181-211. Litman, G. W., Rast, J. P., Shamblott, M. J., Haire, R. N., Hulst, M., Roess, W., Litman, R. T., Hinds-Frey, K. R., Zilch, A., Amemiya, C. T. (1993). Phylogenetic diversification of immunoglobulin genes and the antibody repertoire. Mol Biol Evol, 10(1), 60-72. Manis, J. P., Gu, Y., Lansford, R., Sonoda, E., Ferrini, R., Davidson, L., Rajewsky, K., Alt, F. W. (1998). Ku70 is required for late B cell development and immunoglobulin heavy chain class switching. J Exp Med, 187(12), 2081-2089. Manis, John P., Tian, Ming, & Alt, Frederick W. (2002). Mechanism and control of class-switch recombination. Trends Immunol, 23(1), 31-39. Martin, A., & Scharff, M. D. (2002). Somatic hypermutation of the AID transgene in B and non-B cells. Proc Natl Acad Sci U S A, 99(19), 12304-12308. Martin, Alberto, & Scharff, Matthew D. (2002). AID and mismatch repair in antibody diversification. Nat Rev Immunol, 2(8), 605-614. Min, I. M., Schrader, C. E., Vardo, J., Luby, T. M., D'Avirro, N., Stavnezer, J., & Selsing, E. (2003). The Smu tandem repeat region is critical for Ig isotype switching in the absence of Msh2. Immunity, 19(4), 515-524. Mizuta, R., Iwai, K., Shigeno, M., Mizuta, M., Uemura, T., Ushiki, T., & Kitamura, D. (2003). Molecular visualization of immunoglobulin switch region RNA/DNA complex by atomic force microscope. J Biol Chem, 278(7), 4431-4434. Moldovan, G. L., & D'Andrea, A. D. (2009). How the fanconi anemia pathway guards the genome. Annu Rev Genet, 43, 223-249. Mullen, J. R., Kaliraman, V., Ibrahim, S. S., & Brill, S. J. (2001). Requirement for three novel protein complexes in the absence of the Sgs1 DNA helicase in Saccharomyces cerevisiae. Genetics, 157(1), 103-118. Munoz, I. M., Hain, K., Declais, A. C., Gardiner, M., Toh, G. W., Sanchez-Pulido, L., Heuckmann, J. M., Toth, R., Macartney, T., Eppink, B., Kanaar, R., Ponting, C. P., Lilley, D. M., Rouse, J. (2009). Coordination of structure-specific nucleases by human SLX4/BTBD12 is required for DNA repair. Mol Cell, 35(1), 116-127. Muramatsu, Masamichi, Kinoshita, Kazuo, Fagarasan, Sidonia, Yamada, Shuichi, Shinkai, Yoichi, & Honjo, Tasuku. (2000). Class Switch Recombination and Hypermutation Require Activation-Induced Cytidine Deaminase (AID), a Potential RNA Editing Enzyme. Cell, 102(5), 553-563. Okazaki, I. M., Hiai, H., Kakazu, N., Yamada, S., Muramatsu, M., Kinoshita, K., & Honjo, T. (2003). Constitutive expression of AID leads to tumorigenesis. J Exp Med, 197(9), 1173-1181. Petersen-Mahrt, S. K., Harris, R. S., & Neuberger, M. S. (2002). AID mutates E. coli suggesting a DNA deamination mechanism for antibody diversification. Nature, 418(6893), 99-103. Pham, P., Bransteitter, R., Petruska, J., & Goodman, M. F. (2003). Processive AID-catalysed cytosine deamination on single-stranded DNA simulates somatic hypermutation. Nature, 424(6944), 103-107. Rada, C., Di Noia, J. M., & Neuberger, M. S. (2004). Mismatch recognition and uracil excision provide complementary paths to both Ig switching and the A/T-focused phase of somatic mutation. Mol Cell, 16(2), 163-171. Rada, C., Williams, G. T., Nilsen, H., Barnes, D. E., Lindahl, T., & Neuberger, M. S. (2002). Immunoglobulin isotype switching is inhibited and somatic hypermutation perturbed in UNG-deficient mice. Curr Biol, 12(20), 1748-1755. Ramiro, Almudena R., & Nussenzweig, Michel C. (2004). Immunology: Aid for AID. Nature, 430(7003), 980-981. Reaban, M. E., & Griffin, J. A. (1990). Induction of RNA-stabilized DNA conformers by transcription of an immunoglobulin switch region. Nature, 348(6299), 342-344. Reaban, M. E., Lebowitz, J., & Griffin, J. A. (1994). Transcription induces the formation of a stable RNA.DNA hybrid in the immunoglobulin alpha switch region. J Biol Chem, 269(34), 21850-21857. Revy, Patrick, Muto, Taro, Levy, Yves, Geissmann, Frederic, Plebani, Alessandro, Sanal, Ozden, Catalan, Nadia, Forveille, Monique, Dufourcq-Lagelouse, Remi, Gennery, Andrew, Tezcan, Ilhan, Ersoy, Fugen, Kayserili, Hulya, Ugazio, Alberto G., Brousse, Nicole, Muramatsu, Masamichi, Notarangelo, Luigi D., Kinoshita, Kazuo, Honjo, Tasuku, Fischer, Alain, Durandy, Anne. (2000). Activation-Induced Cytidine Deaminase (AID) Deficiency Causes the Autosomal Recessive Form of the Hyper-IgM Syndrome (HIGM2). Cell, 102(5), 565-575. Rouse, J. (2009). Control of genome stability by SLX protein complexes. Biochem Soc Trans, 37(Pt 3), 495-510. Schrader, C. E., Edelmann, W., Kucherlapati, R., & Stavnezer, J. (1999). Reduced isotype switching in splenic B cells from mice deficient in mismatch repair enzymes. J Exp Med, 190(3), 323-330. Schwartz, E. K., & Heyer, W. D. (2011). Processing of joint molecule intermediates by structure-selective endonucleases during homologous recombination in eukaryotes. Chromosoma, 120(2), 109-127. Seidl, K. J., Bottaro, A., Vo, A., Zhang, J., Davidson, L., & Alt, F. W. (1998). An expressed neo(r) cassette provides required functions of the 1gamma2b exon for class switching. Int Immunol, 10(11), 1683-1692. Shi, H. J., Wen, J. K., Miao, S. B., Liu, Y., & Zheng, B. (2012). KLF5 and hhLIM cooperatively promote proliferation of vascular smooth muscle cells. Mol Cell Biochem, 367(1-2), 185-194. Shinkura, R., Ito, S., Begum, N. A., Nagaoka, H., Muramatsu, M., Kinoshita, K., Sakakibara, Y., Hijikata, H., Honjo, T. (2004). Separate domains of AID are required for somatic hypermutation and class-switch recombination. Nat Immunol, 5(7), 707-712. Stavnezer, J. (1996). Immunoglobulin class switching. Curr Opin Immunol, 8(2), 199-205. Stavnezer, J., Guikema, J. E., & Schrader, C. E. (2008). Mechanism and regulation of class switch recombination. Annu Rev Immunol, 26, 261-292. Stavnezer, Janet, & Schrader, Carol E. (2006). Mismatch repair converts AID-instigated nicks to double-strand breaks for antibody class-switch recombination. Trends Genet, 22(1), 23-28. Stoepker, C., Hain, K., Schuster, B., Hilhorst-Hofstee, Y., Rooimans, M. A., Steltenpool, J., Oostra, A. B., Eirich, K., Korthof, E. T., Nieuwint, A. W., Jaspers, N. G., Bettecken, T., Joenje, H., Schindler, D., Rouse, J., de Winter, J. P. (2011). SLX4, a coordinator of structure-specific endonucleases, is mutated in a new Fanconi anemia subtype. Nat Genet, 43(2), 138-141. Svendsen, J. M., & Harper, J. W. (2010). GEN1/Yen1 and the SLX4 complex: Solutions to the problem of Holliday junction resolution. Genes Dev, 24(6), 521-536. Svendsen, J. M., Smogorzewska, A., Sowa, M. E., O'Connell, B. C., Gygi, S. P., Elledge, S. J., & Harper, J. W. (2009). Mammalian BTBD12/SLX4 assembles a Holliday junction resolvase and is required for DNA repair. Cell, 138(1), 63-77. Ta, V. T., Nagaoka, H., Catalan, N., Durandy, A., Fischer, A., Imai, K., Nonoyama, S., Tashiro, J., Ikegawa, M., Ito, S., Kinoshita, K., Muramatsu, M., Honjo, T. (2003). AID mutant analyses indicate requirement for class-switch-specific cofactors. Nat Immunol, 4(9), 843-848. Takeishi, S., Matsumoto, A., Onoyama, I., Naka, K., Hirao, A., & Nakayama, K. I. (2013). Ablation of Fbxw7 eliminates leukemia-initiating cells by preventing quiescence. Cancer Cell, 23(3), 347-361. Tian, M., & Alt, F. W. (2000). Transcription-induced cleavage of immunoglobulin switch regions by nucleotide excision repair nucleases in vitro. J Biol Chem, 275(31), 24163-24172. Tonegawa, Susumu. (1983). Somatic generation of antibody diversity. Nature, 302(5909), 575-581. Tsai, Albert, & Lieber, Michael. (2010). Mechanisms of chromosomal rearrangement in the human genome. BMC Genomics, 11(Suppl 1), S1. Walker, J. R., Corpina, R. A., & Goldberg, J. (2001). Structure of the Ku heterodimer bound to DNA and its implications for double-strand break repair. Nature, 412(6847), 607-614. Wang, R., Wang, Y., Liu, N., Ren, C., Jiang, C., Zhang, K., Yu, S., Chen, Y., Tang, H., Deng, Q., Fu, C., Wang, Y., Li, R., Liu, M., Pan, W., Wang, P. (2013). FBW7 regulates endothelial functions by targeting KLF2 for ubiquitination and degradation. Cell Res, 23(6), 803-819. Wu, X., Geraldes, P., Platt, J. L., & Cascalho, M. (2005). The double-edged sword of activation-induced cytidine deaminase. J Immunol, 174(2), 934-941. Yoshikawa, K., Okazaki, I. M., Eto, T., Kinoshita, K., Muramatsu, M., Nagaoka, H., & Honjo, T. (2002). AID enzyme-induced hypermutation in an actively transcribed gene in fibroblasts. Science, 296(5575), 2033-2036. Yu, K., Chedin, F., Hsieh, C. L., Wilson, T. E., & Lieber, M. R. (2003). R-loops at immunoglobulin class switch regions in the chromosomes of stimulated B cells. Nat Immunol, 4(5), 442-451. Zarrin, A. A., Tian, M., Wang, J., Borjeson, T., & Alt, F. W. (2005). Influence of switch region length on immunoglobulin class switch recombination. Proc Natl Acad Sci U S A, 102(7), 2466-2470. Zelazowski, P, Max, E E, Kehry, M R, & Snapper, C M. (1997). Regulation of Ku expression in normal murine B cells by stimuli that promote switch recombination. J Immunol, 159(6), 2559-2562. Zhang, J., Bottaro, A., Li, S., Stewart, V., & Alt, F. W. (1993). A selective defect in IgG2b switching as a result of targeted mutation of the I gamma 2b promoter and exon. EMBO J, 12(9), 3529-3537. Zimmerman, R. S., Cox, S., Lakdawala, N. K., Cirino, A., Mancini-DiNardo, D., Clark, E., . . . Funke, B. H. (2010). A novel custom resequencing array for dilated cardiomyopathy. Genet Med, 12(5), 268-278. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17302 | - |
dc.description.abstract | 抗體類型轉換重組 (class switch recombination, CSR) 是B細胞為了有效抵抗外來抗原所進行的分子機制,可使B細胞原分泌IgM轉換成分泌不同類型的抗體。因CSR分子機制相當複雜,以至於目前研究對CSR各階段所參與之因子尚不清楚。故本論文欲探討參與CSR機制中尚未被發掘之潛在重要分子。藉由微陣列 (microarray) 分析小鼠B細胞於刺激進行CSR後表現量上升的基因,挑選出目標基因Slx1。Slx1與Slx4形成複合體,為一結構特異性內切酶 (structure specific endonuclease),能專一性截切基因重組過程中的數種DNA二級結構,依功能及表現量上預測,Slx1有可能參與於CSR反應。 本篇研究目的即是於小鼠B細胞株 (CH12F3) 中,以基因抑制和基因剔除之方式探討Slx1是否作用於CSR。CH12F3細胞經刺激後會進行CSR,自表現IgM轉變為表現IgA的抗體類型。本研究發現,受刺激兩天之細胞其Slx1基因表現量約為未受刺激細胞的兩倍。為驗證Slx1對CSR的重要性,本研究以具有Slx1 shRNA的質體轉染入細胞中,得到一株Slx1基因抑制效率高達80% 之轉染株,其CSR發生頻率為控制組的1.6倍。為更加確定此結果,本論文嘗詴於細胞中剔除Slx1基因。目前已得到Slx1+/puro及Slx1+/-之細胞株,進行分析後發現其CSR發生頻率與控制組相比為顯著性的降低,此表現型與Slx1基因抑制之實驗截然不同。由此研究結果推測Slx1可能參與CSR機制中,但未來仍需更多實驗結果證實Slx1於CSR所扮演的角色。 | zh_TW |
dc.description.abstract | To efficiently defend various antigens, B cells secrete many different isotypes of antibodies. Upon stimulation, B cells switch from secreting IgM to other isotypes of antibodies. The isotype switching is through class switch recombination (CSR). Until now, the detailed molecular mechanism of CSR remains unclear. Therefore, this study aimed to gain insights into CSR by analyzing the differential gene expression pattern from microarray data of the murine B cell line. The up-regulated genes during CSR were further confirmed by qRT-PCR. From literature researching, we selected Slx1 as the potential candidate gene. Slx1 forms the complex with Slx4 and the complex is known as a structure-specific endonuclease. Previous studies indicate that the Slx1-Slx4 complex processes various DNA secondary structures formed during DNA repair. Hence, Slx1 might be involved in CSR.
In this study, through knocking down Slx1 or knocking out Slx1 gene in CH12F3 cells, the CSR model cell line, the role of Slx1 in CSR was investigated. First, Slx1 mRNA level in stimulated CH12F3 cells was about two-fold of unstimulated cells by qRT-PCR assay. The result was similar with the microarray analysis. Moreover, CSR frequency in Slx1 knockdown cells increased comparing to wild type. As for Slx1 knockout in CH12F3 cells (Slx1+/-), surprisingly, the CSR was dramatically reduced comparing to wild type, which was totally opposite to the results of Slx1 knockdown experiments. The results suggest that Slx1 might be involved in CSR. The requirement and the role of Slx1 in CSR need further experiments to confirm. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T00:05:47Z (GMT). No. of bitstreams: 1 ntu-102-R00b22016-1.pdf: 930176 bytes, checksum: f6c4a35ff1134369a8bd934b640152e1 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 謝辭 i
摘要 ii Abstract iii 縮寫表 v 目錄 vi 第一章、緒論 1 1.1 抗體增加變異性之三種分子機制 1 1.2 抗體類型轉換機制 (Class Switch Recombination, CSR) 2 1.2.1變換區 (Switch region, S region) 3 1.2.2 Germ-line transcript (GLT) 3 1.2.3 誘發活化性胞嘧啶核苷脫氨酶 (Activation Induced cytidine Deaminase, AID) 4 1.2.4 鹼基切除修復 (Base Excision Repair, BER) 與核酸配對錯誤修復 (Mismatch Repair, MMR) 6 1.2.5 非同源性黏合 (Non-Homologous End Joining, NHEJ) 7 1.3 Slx1 7 1.3.1 Slx1-Slx4複合體 8 1.3.2 Slx1 8 1.3.3 Slx4 9 1.4 本論文研究之目的與主題 10 第二章、材料與方法 11 2.1 細胞培養與鑑定 11 2.1.1 細胞培養與刺激 11 2.1.2 細胞染色及流式細胞儀分析 11 2.2 基因選殖與質體DNA之建構 11 2.2.1 聚合酶鏈反應 (Polymerase Chain Reaction, PCR) 12 2.2.2 DNA瓊脂糖膠體電泳 12 2.2.3 TA cloning 13 2.2.4 大腸桿菌勝任細胞之製備 13 2.2.5 電轉型作用 13 2.2.6 質體DNA萃取與定序 14 2.2.7 定位點突變 (Site-directed mutagenesis) 14 2.3 基因抑制 (Knock down) 14 2.3.1 shRNA質體之製備 14 2.3.2 轉染作用 15 2.3.3 慢病毒感染 (Lentivirus Infection) 15 2.4 基因剔除 (Knock out) 15 2.4.1 原理與質體之建構 15 2.4.2 基因剔除之質體轉染作用 16 2.4.3 染色體DNA萃取 (酚-氯仿) 16 2.5 RNA之定量 17 2.5.1 抽取total RNA 17 2.5.2 DNase處理 17 2.5.3製備互補DNA (Complementary DNA, cDNA) 18 2.5.4 即時定量聚合酶鏈反應 (Real Time PCR) 18 2.5.5 相對定量 (Relative Quantization, ΔΔCt method) 18 2.6 蛋白質分析 19 2.6.1 常用試劑之製備 19 2.6.2 使用之抗體與廠牌 19 2.6.3 樣品之處理 19 2.6.4 蛋白質膠體電泳 20 2.6.5 蛋白質轉印 20 2.6.6 免疫呈色法 20 第三章、實驗結果 21 3.1 微陣列 (Microarray) 分析 21 3.1.1 候選基因 21 3.1.2 目標基因─Slx1 22 3.2 Slx1於CH12F3細胞之表現量分析 22 3.2.1 Slx1及Slx4之mRNA表現量 22 3.2.2 大腸桿菌過量表現Slx1之系統測試抗體條件 22 3.2.3 CH12F3中Slx1蛋白質表現情形 23 3.3 於CH12F3細胞中抑制Slx1基因表現 23 3.3.1 以shRNA轉染抑制基因表現 23 3.3.2 以慢病毒感染 (Lentivirus infection) 抑制基因表現 24 3.3.3 轉染株之CSR發生頻率 24 3.3.4 轉染株表現型之分子機制分析 24 3.4 建構穩定過量表現Slx1蛋白質之轉染株 25 3.4.1 Slx1補償實驗 (Complementary assay) 26 3.5 Slx1基因剔除 26 3.5.1 Slx1+/puro表現型 26 3.5.2 Slx1+/- 表現型 27 第四章、討論 28 4.1 候選基因 28 4.1.1 Ptgs1 (Prostaglandin-Endoperoxide Synthase 1) 基因 28 4.1.2 Fbxw7 (F-Box And WD Repeat Domain Containing 7) 基因 28 4.1.3 Adamtsl2 (A Disintegrin And Metalloproteinase with Thrombospondin motifs like 2) 基因 29 4.1.4 Csrp3 (Cysteine And Glycine-Rich Protein 3) 基因 29 4.1.5 Ufsp2 (UFM1-Specific Peptidase 2) 基因 29 4.1.6 Fabp3、Slc31a2、9030617O03 Rik基因 30 4.2 Slx1抗體 30 4.3 於細胞中表現外源性Slx1 30 4.4 基因抑制及基因剔除 31 第五章、本研究之未來發展 33 第六章、實驗圖表 34 圖一、受刺激兩天至三天的CH12F3細胞株其Slx1及Slx4之基因表現量 34 圖二、以大腸桿菌過量表現小鼠Slx1蛋白質並測試不同來源之Slx1抗體 35 圖三、分析未受刺激CH12F3細胞中Slx1蛋白質表現之情形 36 圖四、穩定表現Slx1或Slx4 shRNA各轉染株其Slx1或Slx4基因表現情形 37 圖五、含Slx1 shRNA之慢病毒感染所得細胞株其Slx1基因表現情形 38 圖六、穩定表現Slx1 shRNA細胞株其CSR發生頻率 39 圖七、穩定表現Slx1 shRNA轉染株中其germline transcript、AID及Slx4之基因表現情形 40 圖八、於CH12F3細胞株建構穩定過量表現小鼠Slx1之細胞株 41 圖九、Slx1+/puro細胞之Slx1基因表現量 42 圖十、Slx1+/puro細胞株於刺激一至三天之CSR發生頻率 43 圖十一、Slx1+/puro細胞株於刺激一至三天之CSR發生頻率的變化 44 圖十二、Slx1+/-細胞株刺激一天之CSR發生頻率 45 表一、候選基因之文獻資料 46 參考文獻 47 附圖 55 附圖一、穩定表現Ufps2 shRNA細胞株其Ufsp2基因表現量及其CSR發生頻率 55 附圖二、穩定表現Slx1 shRNA細胞株其CSR發生頻率 56 附圖三、穩定表現Slx4 shRNA細胞株其CSR發生頻率 57 附圖四、以大腸桿菌過量表現小鼠Slx1蛋白質並測試Abnova之Slx1抗體 58 附圖五、鑑定Slx1基因剔除細胞株之PCR引子對 59 附錄一、引子序列 60 附錄二、PCR反應條件 62 附錄三、shRNA菌株編號對應之TRCN編號 64 | |
dc.language.iso | zh-TW | |
dc.title | 核酸內切酶Slx1對抗體類型轉換重組之影響及其功能探討 | zh_TW |
dc.title | Studies on the role of Slx1 in class switch recombination | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 周綠蘋(Lu-Ping Chow),黃兆祺(Eric Hwang),陳彥榮(Yen-Rong Chen) | |
dc.subject.keyword | 抗體類型轉換重組,Slx1,Slx4,基因抑制,基因剔除, | zh_TW |
dc.subject.keyword | class switch recombination (CSR),Slx1,Slx4,knockdown,knockout, | en |
dc.relation.page | 64 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2013-08-13 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
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