請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/51012
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 余榮熾 | |
dc.contributor.author | Bi-Shan Liou | en |
dc.contributor.author | 劉碧珊 | zh_TW |
dc.date.accessioned | 2021-06-15T13:23:51Z | - |
dc.date.available | 2019-07-04 | |
dc.date.copyright | 2016-07-04 | |
dc.date.issued | 2016 | |
dc.date.submitted | 2016-06-22 | |
dc.identifier.citation | 1. Mann, J.D., Cahan, A., Gelb, A.G., Fisher, N., Hamper, J., Tippett, P., Sanger, R., and Race, R.R. (1962). A sex-linked blood group. Lancet 1, 8-10.
2. Campana, T., Szabo, P., Piomelli, S., and Siniscalco, M. (1978). The Xga antigen on red cells and fibroblasts. Cytogenet Cell Genet 22, 524-526. 3. Fraser, G.R., Steinberg, A.G., Defaranas, B., Mayo, O., Stamatoyannopoulos, G., and Motulsky, A.G. (1969). Gene frequencies at loci determining blood-group and serum-protein polymorphisms in two villages of northwestern Greece. Am J Hum Genet 21, 46-60. 4. Bhatia, H.M. (1963). Frequency of sex-linked blood group Xg-a in Indians in Bombay. Preliminary study. Indian J Med Sci 17, 491-492. 5. Chown, B., Lewis, M., and Kaita, H. (1964). The Xg Blood Group System: Data on 294 White Families, Mainly Canadian. Can J Genet Cytol 6, 431-434. 6. Devenish, A., Burslem, M.F., Morris, R., and Contreras, M. (1986). Serologic characteristics of a further example of anti-Xga and the frequency of Xga in North London blood donors. Transfusion 26, 426-427. 7. Dewey, W.J., and Mann, J.D. (1967). Xg blood group frequencies in some further populations. J Med Genet 4, 12-15. 8. Gavin, J., Tippett, P., Sanger, R., and Race, R.R. (1963). The Xg Blood Groups of Negroes. Nature 200, 82-83. 9. Ratanaubol, K., and Ratanasirivanich, P. (1971). Xg blood groups of Thais. Nature 229, 430. 10. Sanger, R., Tippett, P., and Gavin, J. (1971). The X-linked blood group system Xg. Tests on unrelated people and families of northern European ancestry. J Med Genet 8, 427-433. 11. Toivanen, P., and Hirvonen, T. (1969). Fetal development of red cell antigens K, k, Lua, Lub, Fya, Fyb, Vel and Xga. Scand J Haematol 6, 49-55. 12. Szabo, P., Campana, T., and Siniscalco, M. (1977). Radioimmune assay for the Xg(a) surface antigen at the individual red cell level. Biochem Biophys Res Commun 78, 655-662. 13. Daniels, G., and Green, C. (2000). Expression of red cell surface antigens during erythropoiesis. Vox Sang 78 Suppl 2, 149-153. 14. Ellis, N.A., Tippett, P., Petty, A., Reid, M., Weller, P.A., Ye, T.Z., German, J., Goodfellow, P.N., Thomas, S., and Banting, G. (1994). PBDX is the XG blood group gene. Nat Genet 8, 285-290. 15. Weller, P.A., Critcher, R., Goodfellow, P.N., German, J., and Ellis, N.A. (1995). The human Y chromosome homologue of XG: transcription of a naturally truncated gene. Hum Mol Genet 4, 859-868. 16. Meynet, O., Scotlandi, K., Pradelli, E., Manara, M.C., Colombo, M.P., Schmid-Antomarchi, H., Picci, P., Bernard, A., and Bernard, G. (2010). Xg expression in Ewing's sarcoma is of prognostic value and contributes to tumor invasiveness. Cancer Res 70, 3730-3738. 17. Goodfellow, P. (1983). Expression of the 12E7 antigen is controlled independently by genes on the human X and Y chromosomes. Differentiation 23 Suppl, S35-39. 18. Ellis, N.A., Ye, T.Z., Patton, S., German, J., Goodfellow, P.N., and Weller, P. (1994). Cloning of PBDX, an MIC2-related gene that spans the pseudoautosomal boundary on chromosome Xp. Nat Genet 6, 394-400. 19. Smith, M.J., Goodfellow, P.J., and Goodfellow, P.N. (1993). The genomic organisation of the human pseudoautosomal gene MIC2 and the detection of a related locus. Hum Mol Genet 2, 417-422. 20. Goodfellow, P., Pym, B., Mohandas, T., and Shapiro, L.J. (1984). The cell surface antigen locus, MIC2X, escapes X-inactivation. Am J Hum Genet 36, 777-782. 21. Berletch, J.B., Yang, F., Xu, J., Carrel, L., and Disteche, C.M. (2011). Genes that escape from X inactivation. Hum Genet 130, 237-245. 22. Levy, R., Dilley, J., Fox, R.I., and Warnke, R. (1979). A human thymus-leukemia antigen defined by hybridoma monoclonal antibodies. Proc Natl Acad Sci U S A 76, 6552-6556. 23. Hahn, J.H., Kim, M.K., Choi, E.Y., Kim, S.H., Sohn, H.W., Ham, D.I., Chung, D.H., Kim, T.J., Lee, W.J., Park, C.K., et al. (1997). CD99 (MIC2) regulates the LFA-1/ICAM-1-mediated adhesion of lymphocytes, and its gene encodes both positive and negative regulators of cellular adhesion. J Immunol 159, 2250-2258. 24. Yoon, S.S., Jung, K.I., Choi, Y.L., Choi, E.Y., Lee, I.S., Park, S.H., and Kim, T.J. (2003). Engagement of CD99 triggers the exocytic transport of ganglioside GM1 and the reorganization of actin cytoskeleton. FEBS Lett 540, 217-222. 25. Pettersen, R.D., Bernard, G., Olafsen, M.K., Pourtein, M., and Lie, S.O. (2001). CD99 signals caspase-independent T cell death. J Immunol 166, 4931-4942. 26. Bernard, G., Breittmayer, J.P., de Matteis, M., Trampont, P., Hofman, P., Senik, A., and Bernard, A. (1997). Apoptosis of immature thymocytes mediated by E2/CD99. J Immunol 158, 2543-2550. 27. Lee, K.J., Yoo, Y.H., Kim, M.S., Yadav, B.K., Kim, Y., Lim, D., Hwangbo, C., Moon, K.W., Kim, D., Jeoung, D., et al. (2015). CD99 inhibits CD98-mediated beta1 integrin signaling through SHP2-mediated FAK dephosphorylation. Exp Cell Res 336, 211-222. 28. Bernard, G., Raimondi, V., Alberti, I., Pourtein, M., Widjenes, J., Ticchioni, M., and Bernard, A. (2000). CD99 (E2) up-regulates alpha4beta1-dependent T cell adhesion to inflamed vascular endothelium under flow conditions. Eur J Immunol 30, 3061-3065. 29. Tippett, P., Shaw, M.A., Green, C.A., and Daniels, G.L. (1986). The 12E7 red cell quantitative polymorphism: control by the Y-borne locus, Yg. Ann Hum Genet 50, 339-347. 30. Goodfellow, P.N., and Tippett, P. (1981). A human quantitative polymorphism related to Xg blood groups. Nature 289, 404-405. 31. Goodfellow, P.J., Pritchard, C., Tippett, P., and Goodfellow, P.N. (1987). Recombination between the X and Y chromosomes: implications for the relationship between MIC2, XG and YG. Ann Hum Genet 51, 161-167. 32. Latron, F., Blanchard, D., and Cartron, J.P. (1987). Immunochemical characterization of the human blood cell membrane glycoprotein recognized by the monoclonal antibody 12E7. Biochem J 247, 757-764. 33. Petty, A.C., and Tippett, P. (1995). Investigation of the biochemical relationship between the blood group antigens Xga and CD99 (12E7 antigen) on red cells. Vox Sang 69, 231-235. 34. Fouchet, C., Gane, P., Cartron, J.P., and Lopez, C. (2000). Quantitative analysis of XG blood group and CD99 antigens on human red cells. Immunogenetics 51, 688-694. 35. Fouchet, C., Gane, P., Huet, M., Fellous, M., Rouger, P., Banting, G., Cartron, J.P., and Lopez, C. (2000). A study of the coregulation and tissue specificity of XG and MIC2 gene expression in eukaryotic cells. Blood 95, 1819-1826. 36. Helmuth, L. (2001). Genome research: map of the human genome 3.0. Science 293, 583-585. 37. Lo, H.S., Wang, Z., Hu, Y., Yang, H.H., Gere, S., Buetow, K.H., and Lee, M.P. (2003). Allelic variation in gene expression is common in the human genome. Genome Res 13, 1855-1862. 38. Morley, M., Molony, C.M., Weber, T.M., Devlin, J.L., Ewens, K.G., Spielman, R.S., and Cheung, V.G. (2004). Genetic analysis of genome-wide variation in human gene expression. Nature 430, 743-747. 39. Shastry, B.S. (2009). SNPs: impact on gene function and phenotype. Methods Mol Biol 578, 3-22. 40. Yan, H., Yuan, W., Velculescu, V.E., Vogelstein, B., and Kinzler, K.W. (2002). Allelic variation in human gene expression. Science 297, 1143. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/51012 | - |
dc.description.abstract | Xga是表現於紅血球上的血型抗原,此抗原是否表現會因個人不同,分成Xg (a+)及Xg (a-);CD99是表現於細胞表面的抗原,而CD99及Xga兩種抗原的表現性具有性別上的差異性。過去的研究已知,CD99的表現量在紅血球上呈現高低的差異,且其在紅血球上表現量的高低跟Xga是否表現有著共同調控的關係。研究指出,當Xg(a+)表現型者時,CD99表現量高;Xg(a-)表現型者,CD99表現量低;但在男性的紅血球上會呈現第三種表現型;特定比例的Xg(a-)男性,CD99卻呈現高表現量。然而Xga及CD99抗原特殊表現型之分子遺傳機制尚未明瞭。
本研究室首先取不同個體之血液的gDNA進行研究,依據其Xga及CD99表現型將其分類成上述三種類型,接著取16人的gDNA以NGS進行性染色體大區域定序,範圍包含XG及CD99兩個基因,並排列出此區域之單一核苷酸多型性(single nucleotide polymorphism,簡稱 SNP)。經由分析結果發現,位於CD99及XG 兩個基因間的SNP rs311103 (G/C) 的基因型吻合三種表現型的分佈,可能對兩者間的調控有很大的關聯性。我們先利用了報導基因法確認了SNP rs311103區域,扮演著enhancer的角色。之後進一步使用程式預測可能與此SNP區域結合呈現親和力差異的轉錄因子,得到GATA family (GATA1~6)、LEF1等轉錄因子的預測。 我們利用K-562細胞可進一步被誘導分化為紅血球性狀做為模型,以及real-time PCR去分析這些轉錄因子在細胞中本身表現量的差異,發現GATA1於K-562細胞分化時表現量增加;接著利用報導基因分析法去做進一步的檢驗,經由分析我們推測GATA1最有可能扮演刺激此SNP的角色。從EMSA實驗中,我們觀察到 GATA1與SNP rs311103基因型為G時可能具有專一性結合。後續實驗我們想利用ChIP進一步證實兩者間的結合作用,並利用報導基因法觀察是否影響兩基因之啟動子以及3C (Chromosome conformation capture) 等實驗更進一步去證實GATA1在SNP rs311103異合子型的細胞株中,對於不同對偶基因型的XG及CD99基因間的表現是否有影響,來證實GATA1轉錄因子扮演差異性調控,因而造成Xga及CD99於紅血球上呈現表現差異之分子機制。 | zh_TW |
dc.description.abstract | Xga is a blood group antigen present on red blood cells (RBCs). The expression of the Xga blood group antigen on red blood cells is different among individuals, subdivided into Xg (a+) and Xg (a-). CD99 molecule is a cell surface glycoprotein. The expression of these two antigens are different in gender. According to previous studies, CD99 is expressed on all human cells, but on red cells, the level of expression shows individual variation. Xg (a+) RBCs all belong to the group of CD99 high expressors and Xg (a-) RBCs belong to the group of CD99 low expressors. Interestingly, some Xg (a-) RBCs from males express high level expression of CD99. However, the molecule genetic mechanism of the special phenotypic correlation between Xga and CD99 antigens is still unclear.
In our investigation, we collected the gDNA from different people, and divided the samples into three groups according to the expression levels of XG and CD99 antigens. We took 16 gDNA samples and sequencing the XG and CD99 gene region by Next Generation Sequencing (NGS). Then we arranged all single-nucleotide polymorphisms (SNPs). Through the data analyzed, we found the SNP rs311103 (G/C), located between the XG gene and the CD99 gene, is associated with the three phenotype groups. In the present study, we aim to demonstrate the transcription factor leading to the different expression levels of the XG and CD99 genes. Several transcription factors, including GATA family (GATA1 to GATA6) and LEF1, were predicted in silico to have different binding affinity to the SNP rs311103 regions with the different genotypes of G and C nucleotides. In further investigation using K-562 erythroleukemia cells as a study model, we found that the expression of the GATA1 gene increased after the K-562 cells differentiated into erythrocytic cells. Ectopic expression of the GATA1 transcription factor also lead to a significant induction of the transcriptional activity in the reporter construct with the SNP rs311103-G form genotype. These result highly suggested that GATA1 may play an important role in activating the XG gene and CD99 gene with the G form genotype at the SNP rs311103 position. In future investigations, we plan to demonstrate whether GATA1 will differentially stimulate the expression the XG gene and CD99 gene with the G form and C form genotype in a SNP heterozygous cell line to substantiate functional role of GATA1 transcription factor in the co-regulation of CD99 and XG blood group phenotypes. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T13:23:51Z (GMT). No. of bitstreams: 1 ntu-105-R03b46031-1.pdf: 1881288 bytes, checksum: 72417a42fa5e9a9e212abc7273c4ab71 (MD5) Previous issue date: 2016 | en |
dc.description.tableofcontents | 口試委員會審定書......................................................................................... i
中文摘要...................................................................................................... ii英文摘要...................................................................................................... iv 縮寫表.......................................................................................................... vi 目錄............................................................................................................ viii 圖目錄......................................................................................................... x 第一章 緒論................................................................................................ 1 1.1 人類XG血型系統................................................................................. 1 1.2 CD99抗原............................................................................................. 2 1.3 Xg Blood Group,CD99 antigen與SNP之關聯.................................... 3 1.3.1 單核苷核酸多型性 (Single nucleotide polymorphism, SNP)........ 3 1.3.2 XG血型和CD99抗原與XGSNP............................................... 3 1.4 研究構想及目的.................................................................................... 4 第二章 實驗材料與方法............................................................................. 6 2.1 細胞培養與分化(Cell Culture) .............................................................. 6 2.2 RNA表現之分析................................................................................... 6 2.3 報導基因分析法(Reporter assay) ..........................................................7 2.4 基因轉殖-脂質體轉染法(Liposome Transfection) .................................. 8 2.5 蛋白質表現分析.................................................................................... 9 2.6 電泳遷移率變動分析 (Electrophoretic Mobility Shift Assay, EMSA) ... 10 第三章 結果.............................................................................................. 12 3.1 SNP rs311103基因型對於CD99抗原表現量之影響........................... 12 3.2 SNP rs311103不同基因型對Xga及CD99抗原表型的連結................ 12 3.3 轉錄因子對XG基因及CD99基因不同SNP基因型的轉錄活性調控關 係........................................................................................................ 13 3.3.1 利用電腦預測何種轉錄因子可能會因SNP rs311103基因型不 同而導致其和基因結合時產生親和性差別............................. 14 3.3.2 分析轉錄因子於血球細胞株中的表現量................................. 14 3.3.3 分析可能具有結合能力差異之轉錄因子對SNP rs311103的核甘 酸G及C基因型之轉錄活性影響............................................ 15 3.3.4 證實GATA1對SNP rs311103不同基因型之轉錄活性影響..... 15 3.4 GATA1與SNP rs311103之間的蛋白質與DNA交互作用(interaction) ............................................................................................................. 16 第四章 討論.............................................................................................. 18 第五章 圖表.............................................................................................. 21參考文獻..................................................................................................... 31 附錄............................................................................................................. 35 圖目錄 圖1. SNP rs311103基因型對照於CD99抗原之表現量............................... 21 圖2. SNP rs311103不同基因型G及C的轉錄活性分析.............................. 22 圖3. 以程式預測可能會和SNP rs311103基因型的不同而導致親和性差別之轉錄因子..................................................................................................... 23 圖4. 可能具有結合能力差異之轉錄因子於血球細胞株中的基因表現量.... 25 圖5. 可能具有結合能力差異之轉錄因子對SNP rs311103不同基因型G及C的轉錄活性分析.......................................................................................... 27 圖6. GATA1、GATA2與GATA3對SNP rs311103不同基因型G及C的轉錄活性分析..................................................................................................... 29 圖7. 轉錄因子GATA1、GATA2、GATA3與SNP rs311103基因型間的蛋白質與DNA交互作用.................................................................................... 30 | |
dc.language.iso | zh-TW | |
dc.title | SNP rs311103於紅血球Xg及CD99抗原表現型
形成之分子機制研究 | zh_TW |
dc.title | Study of the molecule mechanism connecting SNP
rs311103 and the blood group Xg /CD99 phenotype formation | en |
dc.type | Thesis | |
dc.date.schoolyear | 104-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 涂玉青,張?仁,朱善德 | |
dc.subject.keyword | CD99 molecule,XG blood group,single nucleotide polymorphism (SNP), | zh_TW |
dc.relation.page | 39 | |
dc.identifier.doi | 10.6342/NTU201600444 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2016-06-23 | |
dc.contributor.author-college | 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 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-105-1.pdf 目前未授權公開取用 | 1.84 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。