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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張明富 | |
| dc.contributor.author | Chih-Hsien Kao | en |
| dc.contributor.author | 高智賢 | zh_TW |
| dc.date.accessioned | 2021-06-13T15:47:58Z | - |
| dc.date.available | 2010-08-13 | |
| dc.date.copyright | 2008-08-13 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-06-30 | |
| dc.identifier.citation | Asabe, S. I., Tanji, Y., Satoh, S., Kaneko, T., Kimura, K., and Shimotohno, K. (1997). The N-terminal region of hepatitis C virus-encoded NS5A is important for NS4A-dependent phosphorylation. J. Virol. 71: 790-6.
Bartenschlager, R., Ahlborn-Laake, L., Mous, J., and Jacobsen, H. (1993). Nonstructural protein 3 of the hepatitis C virus encodes a serine-type proteinase required for cleavage at the NS3/4 and NS4/5 junctions. J. Virol. 67: 3835-44. Behrens, S. E., Tomei, L., and De Francesco, R. (1996). Identification and properties of the RNA-dependent RNA polymerase of hepatitis C virus. EMBO J. 15: 12-22. Bertolini, L., Lacovacci, S., Ponzetto, A., Gorini, G., Battaglia, M., and Carloni, G. (1993) The human bone-marrow-derived B-cell line CE, susceptible to hepatitis C virus infection. Res. Virol. 144: 281-285 Bertolotti A, Lutz Y, Heard DJ, Chambon P and Tora L (1996) hTAF (II)68, a novel RNA/ssDNA-binding protein with homology to the pro-oncoproteins FUS and EWS is associated with both TFIID and RNA polymerase II. EMBO J. 15:5022–5031. Bradley D. W. (1985). The agents of non-A, non-B viral hepatitis. J. Virol. Meth. 103:07-319. Bretner, M. (2005). Existing and future therapeutic options for hepatitis C virus infection. Acta Biochimica Polonica 52: 57-70. Bukh, J., Purcell, R. H., and Miller, R. H. (1992). Sequence analysis of the 5’ noncoding region of hepatitis C virus. Proc. Natl. Acad. Sci. USA. 89: 4942-4946. Bukh, J., Purcell, R. H., Miller, R. H. (1994). Sequence analysis of the core gene of 14 hepatitis C virus genotypes. Proc. Natl. Acad. Sci. 91: 8239-43. Cai, Z., and Sprent, J. (1996) Influence of antigen dose and costimulation on the primary response of CD8+ T cells in vitro. J. Exp. Med. 183: 2247-2257 Carrère-Kremer, S., Montpellier-Pala, C., Cocquerel, L., Wychowski, C., Penin, F., and Dubuisson, J. (2002). Subcellular localization and topology of the p7 polypeptide of hepatitis C virus. J. Virol. 76: 3720-30. Chang, S. C., Cheng, J. C., Kou, Y. H., Kao, C. H., Chiu, C. H., Wu, H. Y., Chang, M. F. (2000). Roles of the AX(4)GKS and arginine-rich motifs of hepatitis C virus RNA helicase in ATP- and viral RNA-binding activity. J. Virol. 74: 9732-7. Cocchi, F., deVico, A.L., Garzino-Demo, A., Arya, S.K., Gallo, R.C., and Lusso, P. (1995) Identification of Rantes, MIP-1β as the major HIV-suppressive factors produced by CD8+ T cells. Science 270: 1181-1185 Choo, Q. L., Kuo, G., Weiner, A. J., Overby, L. R., Bradley, D. W., and Houghton, M. (1989). Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 244:359-62. Choo, Q. L., Weiner, A. J., Overby, L. R., Kuo, G., Houghton, M., and Bradley, D. W. (1990). Hepatitis C virus: the major causative agent of viral non-A, non-B hepatitis. Br. Med. Bull. 46:423-41. Choo, Q. L., Richman, K. H., Han, J. H., Berger, K., Lee, C., Dong, C., Gallegos, C., Coit, D., Medina-Selby, A., Barr, P. J., Weiner, A. J., Bradley, D. W., Kuo, G., and Houghton, M. (1991). Genetic organization and diversity of the hepatitis C virus. Proc. Natl. Acad. Sci. USA 88: 2451-5. Chung, R. T., and Kaplan, L. M. (1999). Heterogeneous nuclear ribonucleoprotein I (hnRNP-I/PTB) selectively binds the conserved 3' terminus of hepatitis C viral RNA. Biochem. Biophys. Res. Commun. 254: 351-62. Crozat A, Aman P, Mandahl N and Ron D (1993) Fusion of CHOP to a novel RNA-binding protein in human myxoid liposarcoma. Nature. 363:640–644. Egger, D., Wolk, B., Gosert, R., Bianchi, L., Blum, H. E., Moradpour, D., and Bienz, K. (2002). Expression of hepatitis C virus proteins induces distinct membrane alterations including a candidate viral replication complex. J. Virol. 76: 5974-84. El-Hage, N., and Luo, G. (2003). Replication of hepatitis C virus RNA occurs in a membrane-bound replication complex containing nonstructural viral proteins and RNA. J. Gen. Virol. 84: 2761-9. Failla, C., Tomei, L., and De Francesco, R. (1994). Both NS3 and NS4A are required for proteolytic processing of hepatitis C virus nonstructural proteins. J. Virol. 68: 3753-60. Franco, M.A., Tin, C., Rott, L.S., van Cotte, J.L., McGhee, J.R., and Greenberg, H.B. (1997) Evidence for CD8+ T-cell immunity to murine rotavirus in the absence of perforin, fas and gamma interferon. J. Virol. 71: 479-486 Friebe, P., and Bartenschlager, R. (2002). Genetic analysis of sequences in the 3’ nontranslated region of hepatitis C virus that are important for RNA replication. J. Virol. 76: 5326-38. Gale, M. J., Korth, M. J., Tang, N. M., Tan, S. L., Hopkins, D. A., Dever, T. E., Polyak, S. J., Gretch, D. R., and Katze, M. G. (1997). Evidence that hepatitis C virus resistance to interferon is mediated through repression of the PKR protein kinase by the nonstructural 5A protein. Virology 230: 217-27. Gao, L., Aizaki, H., He, J. W., and Lai, M. M. (2004). Interactions between viral nonstructural proteins and host protein hVAP-33 mediate the formation of hepatitis C virus RNA replication complex on lipid raft. J. Virol. 78: 3480-8. Gontarek, R. R., Gutshall, L. L., Herold, K. M., Tsai, J., Sathe, G. M., Mao, J., Prescott, C., and Del Vecchio, A. M. (1999). hnRNP C and polypyrimidine tract-binding protein specifically interact with the pyrimidine-rich region within the 3'NTR of the HCV RNA genome. Nucleic Acids Res. 27:1457-63. Gosert, R., Egger, D., Lohmann, V., Bartenschlager, R., Blum, H. E., Bienz, K., and Moradpour, D. (2003). Identification of the hepatitis C virus RNA replication complex in Huh7 cells harboring subgenomic replicons. J. Virol. 77: 5487-92. Grakoui, A., McCourt, D. W., Wychowski, C., Feinstone, S. M., and Rice, C. M. (1993a). A second hepatitis C virus-encoded proteinase. Proc. Natl. Acad. Sci. USA 90: 10583-7. Grakoui, A., McCourt, D. W., Wychowski, C., Feinstone, S. M., and Rice, C. M. (1993b). Characterization of the hepatitis C virus-encoded serine proteinase: determination of proteinase-dependent polyprotein cleavage sites. J. Virol. 67: 2832-43. Griffin, S. D., Beales, L. P., Clarke, D. S., Worsfold, O., Evans, S. D., Jaeger, J., Harris, M. P., and Rowlands, D. J. (2003). The p7 protein of hepatitis C virus forms an ion channel that is blocked by the antiviral drug, Amantadine. FEBS Lett. 535:34-8. Guidotti, L.G., Ishikawa, T., Hobbs, M.V., Matzke, B., Schreiber, R., and Chisari, F.V. (1996) Intracellular inactivation of the hepatitis B virus by cytotoxic T lymphocytes. Immunity 4: 35-36 Hahm B, Kim YK, Kim JH, Kim TY, Jang SK.(1998) Heterogeneous nuclear ribonucleoprotein L interacts with the 3' border of the internal ribosomal entry site of hepatitis C virus.J Virol.72(11):8782-8. Han, J. H., Shyamala, V., Richman, K. H., Brauer, M. J., Irvine, B., Urdea, M. S., Tekamp-Olson, P., Kuo, G., Choo, Q. L., and Houghton, M. (1991). Characterization of the terminal regions of hepatitis C viral RNA: identification of conserved sequences in the 5' untranslated region and poly(A) tails at the 3' end. Proc. Natl. Acad. Sci. 88:1711-5. Hijikata, M., Kato, N., Ootsuyama, Y., Nakagawa, M., and Shimotohno, K. (1991). Gene mapping of the putative structural region of the hepatitis C virus genome by in vitro processing analysis. Proc. Natl. Acad. Sci. USA 88: 5547-5551. Hijikata, M., Mizushima, H., Akagi, T., Mori, S., Kakiuchi, N., Kato, N., Tanaka, T., Kimura, K., and Shimotohno, K. (1993). Two distinct proteinase activities required for the processing of a putative non-structural precursor protein of hepatitis C virus. J. Virol. 67: 4665-75. Holland, J., Spindler, K., Horodyski, F., Grabau, E., Nichol, S., and VandePol, S. (1982) Rapid evolution of RNA genomes. Science 215: 1577-1579 Huang H, Sun F, Owen DM, Li W, Chen Y, Gale M Jr, Ye J..(2007). Hepatitis C virus production by human hepatocytes dependent on assembly and secretion of very low-density lipoproteins. Proc. Natl. Acad. Sci. USA 104:5848-5853. Ito, T., Tahara, S. M., and Lai, M. M. (1998). The 3'-untranslated region of hepatitis C virus RNA enhances translation from an internal ribosomal entry site. J. Virol. 72(11): 8789-96 Ito, T., and Lai, M. M. (1999). An internal polypyrimidine-tract-binding protein-binding site in the hepatitis C virus RNA attenuates translation, which is relieved by the 3'-untranslated sequence. Virology 254: 288-96. Kaito, M., Watanabe, S., Tsukiyama-Kohara, K., Yamaguchi, K., Kobayashi, Y., Konishi, M., Yokoi, M., Ishida, S., Suzuki, S., and Kohara, M. (1994). Hepatitis C virus particle detected by immunoelectron microscopic study. J. Gen. Virol. 75: 1755-60. Kaneko, T., Tanji, Y., Satoh, S., Hijikata, M., Asabe, S., Kimura, K., and Shimotohno, K. (1994). Production of two phosphoproteins from the NS5A region of hepatitis C viral genome. Biochem. Biophys. Res. Commun. 205: 320-6. Kato T, Matsumura T, Heller T, Saito S, Sapp RK, Murthy K, Wakita T, Liang TJ. (2007) Production of Infectious Hepatitis C Virus of Various Genotypes in Cell Cultures. J. Virol. 81: 4405-4411 Kato T, Wakita T..(2005). Production of infectious hepatitis C virus in cell culture. Uirusu.55:287-296. Kato, N., Hijikata, M., Ootsuyama, Y., Nakagawa, M., Ohkoshi, S., Sugimura,T., and Shimotohno, K. (1990). Molecular cloning of the human hepatitis C virus genome from Japanese patients with non-A, non-B hepatitis. Proc. Natl. Acad. Sci. USA 87: 9524-8. Kato, J., Kato, N., Yoshida, H., Ono-Nita, S. K., Shiratori, Y., and Omata, M. (2002). Hepatitis C virus NS4A and NS4B proteins suppress translation in vivo. J. Med. Virol. 66: 187-99. Kato, N., Yoshida, H., Ono-Nita, S. K., Kato, J., Goto, T., Otsuka, M., Lan, K. H., Matsushima, K., Shiratori, Y., and Omata, M. (2000). Activation of intracellular signaling by hepatitis B and C viruses: C-viral core is the most potent signal inducer. Hepatology 32: 405-12. Kim CS, Seol SK, Song OK, Park JH, Jang SK. (2007). An RNA-binding protein, hnRNP A1, and a scraffold protein, septin 6, facilitate hepatitis C virus replication. J. Virol. 81:3852-3865. Kolykhalov, A. A., Feinstone, S. M., and Rice, C. M. (1996). Identification of a highly conserved sequence element at the 3’ terminus of hepatitis C virus genome RNA. J. Virol. 70: 3363-71. Krecic AM, Swanson MS. (1999). hnRNP complexes: composision, struction, and function. Current Opinion in Cell Biology. 11:363-371. Kuo, G., Choo, Q. L., Alter, H. J., Gitnick, G. L., Redeker, A. G., Purcell, R. H., Miyamura, T., Dienstag, J. L., Alter, M. J., Stevens, C. E., Tegtmeier, G. E., Bonino, F., Colombo, M., Lee, W. S., Kuo, C., Berger, K., Shuster, J. R., Overby, L. R., Bradler, D. W. and Houghton, M. (1989). An assay for circulating antibodies to a major etiologic virus of human non-A, non-B hepatitis. Science 244: 362-4. Lerat, H., Honda, M., Beard, M. R., Loesch, K., Sun, J., Yang, Y., Okuda, M., Gosert, R., Xiao, S. Y., Weinman, S. A., and Lemon, S. M. (2002). Steatosis and liver cancer in transgenic mice expressing the structural and nonstructural proteins of hepatitis C virus. Gastroenterology 122: 352-65. Levrero M.(2006).Viral hepatitis and liver cancer：the case of hepatitis C. Oncogene 25:3834-3847. Lohmann, V., Körner, F., Dobierzewska, A., and Bartenschlager, R. (2001). Mutations in hepatitis C virus RNAs conferring cell culture adaptation. J. Virol. 75: 1437-49. Lohmann, V., Hoffmann, S., Herian, U., Penin, F., and Bartenschlager, R. (2003). Viral and cellular determinants of hepatitis C virus RNA replication in cell culture. J. Virol. 77:3007-19. Lohmann, V., Körner, F., Herian, U., and Bartenschlager, R. (1997). Biochemical properties of hepatitis C virus NS5B RNA-dependent RNA polymerase and identification of amino acid sequence motifs essential for enzymatic activity. J. Virol. 71: 8416-28. Lohmann, V., Korner, F., Koch, J., Herian, U., Theilmann, L., and Bartenschlager, R. (1999). Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line. Science 285:110-3. Machold, R.P., Wiertz, E.J., Jones, T.R., Ploegh, H.L. (1997) The HCMV gene products US11 and US2 differ in their ability to attack allelic forms of murine major histocompatibility complex (MHC) class I heavy chains. J. Exp. Med. 185: 363-366 Martell, M., Esteban, J. I. Quer, J., Genesca, J., Weiner, A., Esteban, R., guardia, J., and Gomez, J. (1992). Hepatitis C virus (HCV) circulates as a population of different but closely related genomes: quasispecies nature of HCV genome distribution. J. Virol. 66: 3225-3229. Mercer, D. F., Schiller, D. E., Elliott, J. F., Douglas, D. N., Hao, C., Rinfret, A., Addison, W. R., Fischer, K. P., Churchill, T. A., Lakey, J. R., Tyrrell, D. L., and Kneteman, N. M. (2001). Hepatitis C virus replication in mice with chimeric human livers. Nat. Med. 7:927-33. Moradpour, D., and Blum, H. E. (2004). A primer on the molecular virology of hepatitis C. Liver Int. 24: 519-25 Ogata, N., alter, H. J., Miller, R. H., and Purcell, R. H. (1991). Nucleotide sequence and mutation rate of the H strain of hepatitis C virus. Proc. Natl. Acad. Sci. USA 88: 3392-3396. Okamoto, H., Kojima, M., Okada, S.-I., Yoshizawa, H., Iizuka, H., Tanaka, T., Muchmore, E.E., Peterson, D.A., Ito, Y., and Mishiro, S. (1992) Genetic drift of hepatitis C virus during an 8.2-year infection in chimpanzee : variability and stability. Virology 190: 894-899 Pavic, I., Polic, B., Crnkovic, I., Lucin, P., Jonjic, S., and Koszinowki, U.H. (1993) Participation of endogenous tumor necrosis factor α in host resistance to cytomegalovirus infection. J. Gen. Virol. 74: 2215-2223 Pavlovic, D., Neville, D. C., Argaud, O., Blumberg, B., Dwek, R. A., Fischer, W. B., and Zitzmann, N. (2003). The hepatitis C virus p7 protein forms an ion channel that is inhibited by long-alkyl-chain iminosugar derivatives. Proc. Natl. Acad. Sci. 100: 6104-8. Pflugheber, J., Fredericksen, B., Sumpter Jr., R., Wang, C., Ware, F., Sodora, D. L., and Gale Jr., M. (2002). Regulation of PKR and IRF-1 during hepatitis C virus RNA replication. Proc. Natl. Acad. Sci. USA 99: 4650-5. Piccininni, S., Varaklioti, A., Nardelli, M., Dave, B., Raney, K. D., and McCarthy, J. E. (2002). Modulation of the hepatitis C virus RNA-dependent RNA polymerase activity by the non-structural (NS) 3 helicase and the NS4B membrane protein. J. Biol. Chem. 277: 45670-9. Plagemann, P. G. W. (1991). Hepatitis C virus Arch. Virol. 120: 165-180. Santolini, E., Migliacio, G., and La Monica, N. (1994). Biosynthesis and bio- chemical properties of the hepatitis C virus core protein. J. Virol. 68: 3631-41. Shi, S. T., Lee, K. J., Aizaki, H., Hwang, S. B., and Lai, M. M. (2003). Hepatitis C virus RNA replication occurs on a detergent-resistant membrane that cofractionates with caveolin-2. J. Virol. 77: 4160-8. Shimizu, Y.K., Iwamoto, A., Hijikata, M., Purcell, R.H., Yoshikaru, H. (1992) Evidence for in vitro replication of hepatitis C virus genome in a human T cell line. Proc. Natl. Acad. Sci. USA 89: 5477-5481 Suzuki T, Ishii K, Aizaki H, Wakita T.(2007). Hepatitis C viral life cycle. Adv Drug Deliv Rev. 59:1200-1212. Tai, C. L., Chi, W. K., Chen, D. S., and Hwang, L. H. (1996). The helicase activity associated with hepatitis C virus nonstructural protein 3 (NS3). J. Virol. 70: 8477-84. Takamizawa, A., Mori, C., Fuke, I., Manabe, S., Murakami, S., Fujita, J., Onishi, E., Andoh, T., Yoshida, I., and Okayama, H. (1991). Structure and organization of the hepatitis C virus genome isolated from human carriers. J. Virol. 65: 1105-13. Tan, S. L., and Katze, M. G. (2001). How hepatitis C virus counteracts the interferon response: the jury is still out on NS5A. Virology 284: 1-12. Tan, S. L., Pause, A., Shi, Y., and Sonenberg, N. (2002). Hepatitis C therapeutics: current status and emerging strategies. Nat. Rev. Drug Discov. 1:867-81. Tanji, Y., Kaneko, T., Satoh, S., and Shimotohno, K. (1995). Phosphorylation of hepatitis C virus-encoded nonstructural protein NS5A. J. Virol. 69: 3980-6. Walewski, J. L., Keller, T. R., Stump, D. D., and Branch, A. D. (2001). Evidence for a new hepatitis C virus antigen encoded in an overlapping reading frame. RNA 7: 710-21. Weiner, A. J., Brauer, M. J., Rosenblatt, J., Richman, K. H., Tung, J., Crawford, K., Bonino, F., Saracco, G., Choo, Q. L., Houghton, M., and Han, J. H. (1991). Variable and hypervariable domains are found in the regions of HCV corresponding to the flavivirus envelope and NS1 proteins and the pestivirus envelope glycoproteins. Virology 180:842-8. Weiner, A.J., Geysen, H.M., Christopherson, C., Hall, J.E., Mason, T.J., Saracw, G., Bonine, F., Crawford, K., Marion, C.D., Crawford, K.A., Brunetto, M., Barr, P.J., Miyamura, T., McHutchison, J.G., and Houghtonm M. (1992) Evidence for immune selection of hepatitis C virus (HCV) putative envelope glycoprotein variants: potential role in chronic HCV infection. Proc. Natl. Acad. Sci. USA 89: 3468-3474 Xie, Z. C., Riezu, J. I., Lasarte, J. J., Guillen, J., Su, J. H., Civeira, M. P., and Prieto, J. (1998). Transmission of hepatitis C virus infection to tree shrews. Virology 244: 513-20. Xu, Z., Choi, J., Yen, T. S. B., Lu, W., Strohecker, A., Govindarajan, S., Chien, D., Selby, M. J., and Ou, J. (2001). Synthesis of a novel hepatitis C virus protein by ribosomal frameshift. EMBO J. 20: 3840-8. Yi M, Lemon SM.(2003). 3' nontranslated RNA signals required for replication of hepatitis C virus RNA. J Virol.77:3557-68. You, S., Stump, D. D., Branch, A. D., and Rice, C. M. (2004). A cis-acting replication element in the sequence encoding the NS5B RNA-dependent RNA polymerase is required for hepatitis C virus RNA replication. J. Virol. 78: 1352-66. Zinszner H, Albalat R and Ron D (1994) A novel effector domain from the RNA-binding protein TLS or EWS is required for oncogenic transformation by CHOP. Genes Dev. 8: 2513–2526. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/37863 | - |
| dc.description.abstract | C型肝炎病毒（hepatitis C virus, HCV）屬於黃熱病毒科（Flaviviridae）正向單股RNA病毒。基因體約9.6 kb，能夠產生結構蛋白質core、E1、E2 、p7以及非結構蛋白質NS2、NS3、NS4A、NS4B、NS5A、NS5B。由於C型肝炎病毒RNA複製時的突變率很高，使C型肝炎病毒能逃過宿主的攻擊。C型肝炎病毒主要是經由血液或體液感染，感染後約有80％的病患換轉變成慢性肝炎，其中約有10-20％病患會產生肝硬化，也有約1-5％的病患會轉變為肝癌。全球約有超過1.7億的人口感染C型肝炎病毒，已成為了一個必須積極去面對與解決的全球性健康問題。
本實驗室希望能夠建立一個利用Huh7細胞產生C型肝炎病毒的穩定表現系統。本研究首先將質體pJFH1進行改造，在C型肝炎病毒基因後端再接上D型肝炎病毒的ribozyme和poly A signal，完成質體pJFH1-RZ-PA的建構。將質體pJFH1-RZ-PA轉染到Huh7細胞中，兩天後萃取細胞內的RNA進行RT-PCR，結果能夠偵測到反股的C型肝炎病毒RNA，也能偵測到核心蛋白質的表現，顯示藉由轉染送入的質體所產生的C型肝炎病毒正股RNA能夠自行複製，產生反股RNA。再將質體的T7驅動子置換成CMV 驅動子，完成質體pCMV-JFH1-RZ-PA之建構，再用質體pCMV-JFH1-RZ-PA轉染Huh7細胞，在G418存在下，挑選出能夠穩定表現HCV RNA的Huh7細胞株Huh7-CMV9。 由先前的論文指出，hnRNP（heterogeneous nuclear ribonucleoprotein） family的成員會和C型肝炎病毒RNA結合，幫助C型肝炎病毒的複製。我們利用shRNA技術以及穩定表現HCV RNA的Huh7細胞株Huh7-CMV9，研究hnRNP哪一些成員參與C型肝炎病毒的複製。結果發現 hnRNP P2/ FUS若被knockdown，會使細胞中的HCV反股RNA量減少，顯示hnRNP P2可能參與C型肝炎病毒的複製。 為了尋找抗C型肝炎病毒藥物，利用Huh7-CMV9複製系統以測試藥物對進行C型肝炎病毒於細胞中所累積的反股RNA量的影響。結果顯示從48種藥物之中有10種藥物可抑制C型肝炎病毒於細胞中所累積的反股RNA量一半以上。進一步實驗結果測得藥物D46的IC50濃度為3.43μM。hnRNP P2/ FUS和藥物D46如何調節C型肝炎病毒RNA複製之詳細作用機轉，須再進一步研究。 | zh_TW |
| dc.description.abstract | Hepatitis C virus (HCV) infects about 170 million people worldwide and has been a major public health problem. HCV contains a positive-stranded RNA genome of 9.6 kilobases that encodes a polyprotein of about 3000 amino acid residues. The majority of HCV-infected patients fail to clear the virus and develop chronic liver diseases including cirrhosis and hepatocellular carcinoma. For screening chemicals and cellular factors that can inhibit HCV replication, an infectious HCV clone was first established, plasmid pCMV-JFH1-RZ-PA that contains sequences representing the full-length genomic RNA of HCV under the control of CMV promoter, followed by the ribozyme of hepatitis delta virus was constructed. Huh7 cells were transfected with pCMV-JFH1-RZ-PA and pCMV-Tag2 containing neomycin-resistant gene. Following G418 selection, one clone, designated Huh7-CMV9, that is capable of producing viral RNA and proteins at high levels was identified. In addition, HCV-like particles were detected in the Huh7-CMV9 cells by Electron Microscopy. Heterogeneous nuclear ribonucleoprotein A1 was recently shown to be important for HCV replication. By introducing shRNAs into the Huh7-CMV9 cells in this study, hnRNP P2 was identified to specifically reduce the replication of HCV. On the other hand, 10 out of 48 chemicals examined showed inhibitory effects on HCV replication in the Huh7-CMV9 system. Among these chemicals, the IC50 of chemical D46 to inhibit HCV replication is 3.43μM. How hnRNP P2 and D46 regulate HCV replication requires further studies. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T15:47:58Z (GMT). No. of bitstreams: 1 ntu-97-R94442002-1.pdf: 1804232 bytes, checksum: 095e8364c1209749d624c9a0f4945b93 (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | 中文摘要................. Ⅰ
英文摘要.................. Ⅲ 縮寫表.................... Ⅳ 緒論........................1 實驗材料來源...............13 實驗方法...................17 實驗結果...................28 討論.......................32 圖表.......................36 參考文獻...................55 | |
| dc.language.iso | zh-TW | |
| dc.subject | C型肝炎病毒 | zh_TW |
| dc.subject | 複製 | zh_TW |
| dc.subject | hnRNP | en |
| dc.subject | HCV | en |
| dc.subject | replication | en |
| dc.title | 利用新建立之C型肝炎病毒感染細胞株篩選可調節病毒複製之宿主細胞因子及化合物 | zh_TW |
| dc.title | Selection of host factors and chemicals that regulate the replication of hepatitis C virus by using a newly established infectious clone | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 董馨蓮,李芳仁 | |
| dc.subject.keyword | C型肝炎病毒,複製, | zh_TW |
| dc.subject.keyword | HCV,replication,hnRNP, | en |
| dc.relation.page | 63 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2008-06-30 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 生物化學暨分子生物學研究所 | zh_TW |
| 顯示於系所單位： | 生物化學暨分子生物學科研究所 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-97-1.pdf 未授權公開取用 | 1.76 MB | Adobe PDF |
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