請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60400完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張繼堯(Chi-Yao Chang) | |
| dc.contributor.author | Yi-Sen Wang | en |
| dc.contributor.author | 王奕森 | zh_TW |
| dc.date.accessioned | 2021-06-16T10:17:14Z | - |
| dc.date.available | 2020-08-16 | |
| dc.date.copyright | 2013-08-25 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-08-17 | |
| dc.identifier.citation | 曾文陽。石斑魚養殖學。前程出版社，1998年。
Brash, D. E., Ziegler, A., Jonason, A. S., Simon, J. A., Kunala, S., & Leffell, D. J. (1996). Sunlight and sunburn in human skin cancer: p53, apoptosis, and tumor promotion. J Investig Dermatol Symp Proc, 1(2), 136-142. Castillo, J. P., & Kowalik, T. F. (2002). Human cytomegalovirus immediate early proteins and cell growth control. Gene, 290(1-2), 19-34. Castillo, J. P., Yurochko, A. D., & Kowalik, T. F. (2000). Role of human cytomegalovirus immediate-early proteins in cell growth control. J Virol, 74(17), 8028-8037. Chelbi-Alix, M. K., Quignon, F., Pelicano, L., Koken, M. H., & de The, H. (1998). Resistance to virus infection conferred by the interferon-induced promyelocytic leukemia protein. J Virol, 72(2), 1043-1051. Chi, S. C., Lo, C. F., Kou, G. H., Chang, P. S., Peng, S. E., and Chen, S. N.(1997). Mass mortalities associated with viral nervous necrosis (VNN) disease in two species of hatchery-reared grouper, Epinephelus fuscogutatus and Epinephelus akaara (Temminck & Schlegel). Journal of Fish Diseases 20(3), 185-193. Chou, H. Y., Hsu, C. C., & Peng, T. Y. (1998). Isolation and characterization of a pathogenic iridovirus from cultured grouper (Epinephelus sp.) in Taiwan. Fish Pathology, 33(4), 201-206. De Kinkelin, P., Bearzotti, M. . (1981). Immunization of rainbow trout against viral hemorrhagic septicemia (VHS) with a thermoresistant variant of the virus. . Dev Biol Stand 49, 431-439. Delhalle, S., Duvoix, A., Schnekenburger, M., Morceau, F., Dicato, M., & Diederich, M. (2003). An introduction to the molecular mechanisms of apoptosis. Ann N Y Acad Sci, 1010, 1-8. Delhon, G., Tulman, E. R., Afonso, C. L., Lu, Z., Becnel, J. J., Moser, B. A., Rock, D. L. (2006). Genome of invertebrate iridescent virus type 3 (mosquito iridescent virus). J Virol, 80(17), 8439-8449. doi: 10.1128/JVI.00464-06 Do, J. W., Moon, C. H., Kim, H. J., Ko, M. S., Kim, S. B., Son, J. H., Park, J. W. (2004). Complete genomic DNA sequence of rock bream iridovirus. Virology, 325(2), 351-363. doi: 10.1016/j.virol.2004.05.008 Essani K, Granoff A. . (1989). Amphibian and piscine iridoviruses proposal for nomenclature and taxonomy based on molecular and biological properties. . Intervirology., 30, 187-193. Goorha, R. . (1995). Family Iridoviridae. In: Murphy, F.A., Fauquet, C.M., Bishop, D.H.L., Ghabrial, S.A., Jarvis, A.W., Martelli, G.P. et al. (Eds.), Virus taxonomy: classification and nomenclature of viruses. Sixth report of the international committee on taxonomy of viruses. Arch Virol Suppl 10, 95-99. Gupta, S., Agrawal, A., Agrawal, S., Su, H., & Gollapudi, S. (2006). A paradox of immunodeficiency and inflammation in human aging: lessons learned from apoptosis. Immun Ageing, 3, 5. doi: 10.1186/1742-4933-3-5 He, J. G., Lu, L., Deng, M., He, H. H., Weng, S. P., Wang, X. H., Zhou, S. Y., Long, Q. X., Wang, X. Z., and Chan, S. M. . (2002). Sequence analysis of the complete genome of an iridovirus isolated from the tiger frog. . Virology, 292(2), 185-197. Herbein G, O'Brien WA. (2000). Tumor necrosis factor (TNF)-alpha and TNF receptors in viral pathogenesis. Proc Soc Exp Biol Med., 223(3). Herbein, G., & O'Brien, W. A. (2000). Tumor necrosis factor (TNF)-alpha and TNF receptors in viral pathogenesis. Proc Soc Exp Biol Med, 223(3), 241-257. Hirono, I., Nam, B. H., Kurobe, T., & Aoki, T. (2000). Molecular cloning, characterization, and expression of TNF cDNA and gene from Japanese flounder Paralychthys olivaceus. J Immunol, 165(8), 4423-4427. Horiuchi, T., Mitoma, H., Harashima, S., Tsukamoto, H., & Shimoda, T. (2010). Transmembrane TNF-alpha: structure, function and interaction with anti-TNF agents. Rheumatology (Oxford), 49(7), 1215-1228. doi: 10.1093/rheumatology/keq031 Huang, Y., Huang, X., Liu, H., Gong, J., Ouyang, Z., Cui, H., Qin, Q. (2009). Complete sequence determination of a novel reptile iridovirus isolated from soft-shelled turtle and evolutionary analysis of Iridoviridae. BMC Genomics, 10, 224. doi: 10.1186/1471-2164-10-224 Jacobs, B. L., Jancovich, J. K., Bremont, M., and Touchman, J. W.(2010). Evidence for Multiple Recent Host Species Shifts among the Ranaviruses (Family Iridoviridae). Journal of Virology 84(6), 2636-2647. Jakob, N. J., Kleespies, R. G., Tidona, C. A., Muller, K., Gelderblom, H. R., & Darai, G. (2002). Comparative analysis of the genome and host range characteristics of two insect iridoviruses: Chilo iridescent virus and a cricket iridovirus isolate. J Gen Virol, 83(Pt 2), 463-470. Jakob, N. J., Muller, K., Bahr, U., & Darai, G. (2001). Analysis of the first complete DNA sequence of an invertebrate iridovirus: coding strategy of the genome of Chilo iridescent virus. Virology, 286(1), 182-196. doi: 10.1006/viro.2001.0963 Jancovich, J. K., Mao, J. H., Chinchar, V. G., Wyatt, C., Case, S. T., Kumar, S., Valente, G., Subramanian, S., Davidson, E. W., Collins, J. P., and Jacobs, B. L. . (2003). Genomic sequence of a ranavirus (family Iridoviridae) associated with salamander mortalities in North America. Virology 316(1), 90-103. Kerr, J. F., Wyllie, A. H., & Currie, A. R. (1972). Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer, 26(4), 239-257. Kinchington, P. R., Hougland, J. K., Arvin, A. M., Ruyechan, W. T., & Hay, J. (1992). The varicella-zoster virus immediate-early protein IE62 is a major component of virus particles. J Virol, 66(1), 359-366. Kroemer, G., Dallaporta, B., & Resche-Rigon, M. (1998). The mitochondrial death/life regulator in apoptosis and necrosis. Annu Rev Physiol, 60, 619-642. doi: 10.1146/annurev.physiol.60.1.619 Lam, F. W., Wu, S. Y., Lin, S. J., Lin, C. C., Chen, Y. M., Wang, H. C., Lin, J. H. (2011). The expression of two novel orange-spotted grouper (Epinephelus coioides) TNF genes in peripheral blood leukocytes, various organs, and fish larvae. Fish Shellfish Immunol, 30(2), 618-629. doi: 10.1016/j.fsi.2010.12.011 Lin, P. W., Huang, Y. J., John, J. A., Chang, Y. N., Yuan, C. H., Chen, W. Y., Chang, C. Y. (2008). Iridovirus Bcl-2 protein inhibits apoptosis in the early stage of viral infection. Apoptosis, 13(1), 165-176. doi: 10.1007/s10495-007-0152-y Loparev, V. N., Parsons, J. M., Knight, J. C., Panus, J. F., Ray, C. A., Buller, R. M., Esposito, J. J. (1998). A third distinct tumor necrosis factor receptor of orthopoxviruses. Proc Natl Acad Sci U S A, 95(7), 3786-3791. Lu, L., Zhou, S. Y., Chen, C., Weng, S. P., Chan, S. M., & He, J. G. (2005). Complete genome sequence analysis of an iridovirus isolated from the orange-spotted grouper, Epinephelus coioides. Virology, 339(1), 81-100. doi: 10.1016/j.virol.2005.05.021 Ma, X. (2001). TNF-alpha and IL-12: a balancing act in macrophage functioning. Microbes Infect, 3(2), 121-129. Martinez, F. P., Cosme, R. S., & Tang, Q. (2010). Murine cytomegalovirus major immediate-early protein 3 interacts with cellular and viral proteins in viral DNA replication compartments and is important for early gene activation. J Gen Virol, 91(Pt 11), 2664-2676. doi: 10.1099/vir.0.022301-0 Murali, S., Wu, M. F., Guo, I. C., Chen, S. C., Yang, H. W., and Chang, C. Y. (2002). Molecular characterization and pathogenicity of a grouper iridovirus (GIV) isolated from yellow grouper, Epinephelus awoara (Temminck & Schlegel). . Journal of Fish Diseases 25(2), 91-100. Murata, T., Isomura, H., Yamashita, Y., Toyama, S., Sato, Y., Nakayama, S., Tsurumi, T. (2009). Efficient production of infectious viruses requires enzymatic activity of Epstein-Barr virus protein kinase. Virology, 389(1-2), 75-81. doi: 10.1016/j.virol.2009.04.007 Qin, Q. W., Lam, T. J., Sin, Y. M., Shen, H., Chang, S. F., Ngoh, G. H., & Chen, C. L. (2001). Electron microscopic observations of a marine fish iridovirus isolated from brown-spotted grouper, Epinephelus tauvina. J Virol Methods, 98(1), 17-24. Schultz, D. R., & Harrington, W. J., Jr. (2003). Apoptosis: programmed cell death at a molecular level. Semin Arthritis Rheum, 32(6), 345-369. doi: 10.1053/sarh.2003.50005 Sheikh, M. S., Antinore, M. J., Huang, Y., & Fornace, A. J., Jr. (1998). Ultraviolet-irradiation-induced apoptosis is mediated via ligand independent activation of tumor necrosis factor receptor 1. Oncogene, 17(20), 2555-2563. doi: 10.1038/sj.onc.1202292 Song, W. J., Oin, Q. W., Qiu, J., Huang, C. H., Wang, F., and Hew, C. L. . (2004). Functional Genomics analysis of Singapore grouper iridovirus: Complete sequence determination and proteomic analysis. . Journal of Virology, 78(22), 12576-12590. Song, Y. J., & Stinski, M. F. (2005). Inhibition of cell division by the human cytomegalovirus IE86 protein: role of the p53 pathway or cyclin-dependent kinase 1/cyclin B1. J Virol, 79(4), 2597-2603. doi: 10.1128/JVI.79.4.2597-2603.2005 Stinski, M. F., & Petrik, D. T. (2008). Functional roles of the human cytomegalovirus essential IE86 protein. Curr Top Microbiol Immunol, 325, 133-152. Tan, W. G. H., Barkman, T. J., Chinchar, V. G., and Essani, K. . (2004). Comparative genomic analyses of frog virus 3, type species of the genus Ranavirus (family Iridoviridae). . Virology 323(1), 70-84. Tidona, C. A., & Darai, G. (1997). Molecular anatomy of lymphocystis disease virus. Arch Virol Suppl, 13, 49-56. Tsai, C. T., Lin, C. H., & Chang, C. Y. (2007). Analysis of codon usage bias and base compositional constraints in iridovirus genomes. Virus Res, 126(1-2), 196-206. doi: 10.1016/j.virusres.2007.03.001 Tsai, C. T., Ting, J. W., Wu, M. H., Wu, M. F., Guo, I. C., & Chang, C. Y. (2005). Complete genome sequence of the grouper iridovirus and comparison of genomic organization with those of other iridoviruses. Journal of Virology, 79(4), 2010-2023. doi: Doi 10.1128/Jvi.79.4.2010-2023.2005 van Oers, M. M., Ince, I. A., Westenberg, M., Vlak, J. M., Demirbag, Z., and Nalcacioglu, R. (2008). Open reading frame 193R of Chilo iridescent virus encodes a functional inhibitor of apoptosis (IAP). . Virology 376(1), 124-131. Williams, T. (1996). The iridoviruses. Adv Virus Res, 46, 345-412. Willis, D. B., & Granoff, A. (1985). trans activation of an immediate-early frog virus 3 promoter by a virion protein. J Virol, 56(2), 495-501. Wu, M. S., Chen, C. W., Liu, Y. C., Huang, H. H., Lin, C. H., Tzeng, C. S., & Chang, C. Y. (2012). Transcriptional analysis of orange-spotted grouper reacting to experimental grouper iridovirus infection. Dev Comp Immunol, 37(2), 233-242. doi: 10.1016/j.dci.2012.04.002 Xia, L., Cao, J., Huang, X., & Qin, Q. (2009). Characterization of Singapore grouper iridovirus (SGIV) ORF086R, a putative homolog of ICP18 involved in cell growth control and virus replication. Arch Virol, 154(9), 1409-1416. doi: 10.1007/s00705-009-0457-y Xia, L., Liang, H., Huang, Y., Ou-Yang, Z., & Qin, Q. (2010). Identification and characterization of Singapore grouper iridovirus (SGIV) ORF162L, an immediate-early gene involved in cell growth control and viral replication. Virus Res, 147(1), 30-39. doi: 10.1016/j.virusres.2009.09.015 Zhang, Q. Y., Zhao, Z., Xiao, F., Li, Z. Q., and Gui, J. F. . (2006). Molecular characterization of three Rana grylio virus (RGV) isolates and Paralichthys olivaceus lymphocystis disease virus (LCDV-C) in iridoviruses. . Aquaculture 251(1), 1-10. Zou, J., Wang, T., Hirono, I., Aoki, T., Inagawa, H., Honda, T., Secombes, C. J. (2002). Differential expression of two tumor necrosis factor genes in rainbow trout, Oncorhynchus mykiss. Dev Comp Immunol, 26(2), 161-172. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60400 | - |
| dc.description.abstract | 石斑魚(Epinephelus spp.)是具有高度經濟價值的養殖魚種，在台灣，除了東南亞及中國大陸的養殖興起，造成的經濟競爭外，其受到虹彩病毒的感染，造成魚隻死亡，更是各國都面臨的棘手問題，因此研究虹彩病毒為當前的重要課題。本實驗室藉由cDNA微陣列晶片，分析石斑魚虹彩病毒(grouper iridovirus, GIV)基因表現次序，推測其具有二十一個極早期基因(immediate early gene, IE)，這些基因能藉由調控病毒表現及改變宿主的生理狀態，使病毒得以進行複製增殖。而石斑魚虹彩病毒030L就被偵測為極早期基因，其全長為696個鹼基對，由231個胺基酸組成，蛋白質分子量為25.9 kDa，再經由NCBI資料庫分析GIV030L開放閱讀框架的結果顯示，其為腫瘤壞死因子受體(tumor necrosis factor receptor, TNFR)超級家族的一員。此TNFR蛋白具有膜外TNF配體結合區和穿膜區，卻缺少了膜內功能區，推測此TNFR-like基因可能是虹彩病毒為了對抗寄主TNF誘發系統，所演化出來的圈套。
本實驗建構了真核表現載體pcDNA3CF-030L，轉染於子宮頸部腫瘤(HeLa)細胞中，經紫外光照射後，藉由免疫細胞化學染色及DNA斷裂端標記試驗(TUNEL assay)，發現過量表現的GIV030L無法阻擋紫外光所造成的細胞凋亡，紫外光照射所誘發的細胞凋亡是經由內部路徑(intrinsic pathway)所造成。因此本實驗進一步想藉由TNF-α細胞凋亡外部路徑(extrinsic pathway)來誘發石斑魚腎臟(GK)細胞的細胞凋亡。接著本實驗建構石斑魚TNF-α基因於原核表現載體pET-43.1a(+)-Factor Xa-TNF-α，並成功地在大腸桿菌BL-21(DE3) plysS以IPTG誘導表現出可溶性的重組蛋白，藉由親和性層析管柱，以不同濃度之咪唑(imidazole)沖提出目標蛋白，再由Factor Xa蛋白質切割酶切割後，得到重組的TNF-α蛋白。以不同濃度之TNF-α蛋白，與相同劑量之Actinomycin D處理於石斑魚腎臟細胞中，在顯微鏡下進行觀察，得知隨著TNF-α重組蛋白濃度的增加，GK細胞凋亡的情形也逐步惡化。 | zh_TW |
| dc.description.abstract | Grouper (Epinephelus spp.) are highly economic value farmed fish in Taiwan. In addition to the farming competition from China and Southeast Asia countries, the high mortality caused by iridovirus infection became one of the thorniest issues in grouper aquaculture. Therefore, the study of mechanism involving in the iridovirus infection is the most important current events. Using cDNA microarray assay, our laboratory had identified 21 immediate early genes from grouper iridovirus including GIV030L. The GIV030L contains 696 nucleotides and is composed of 231 amino acids which encode a 25.9 kDa protein. According NCBI database comparative sequence analysis, GIV030L represents a tumor necrosis factor receptor (TNFR) homolog with a TNF ligand binding extracellular domain and a transmembrane domain, but lacking a intracellular domain. This incomplete TNFR-like viral protein may serve as a decoy and play a role to help iridovirus against host TNF attacking system.
In this study, GIV030L was constructed into pcDNA3CF eukaryotic expression vector and was transfected into HeLa cells. However, the overexpressed GIV030L can not prevent the HeLa cells from apoptosis trigged by UV-irradiation mediated intrinsic apoptosis pathway in the immunocytochemistry and Terminal deoxynucleotidyl Transferase mediated dUTP Nick End Labeling (TUNEL) assays. To establish the grouper kidney (GK) cells apoptosis through TNF and TNFR extrinsic pathway, the TNF-α gene was cloned from orange-spotted grouper and was constructed into pET-43.1a(+)-Factor Xa prokaryotic expression vector. The E. coli expressed recombinant TNF-α contains a NusA protein at N-terminal to help TNF-α under correct folding. After Ni2+ affinity column purification and Factor Xa protease digestion, the soluble recombinant grouper TNF-α was subjected to apoptosis experiment. With no harmful concentration of Actinomycin D, the GK cells were gradually deteriorated fallowing the increase of recombinant TNF-α addition. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T10:17:14Z (GMT). No. of bitstreams: 1 ntu-102-R00b45018-1.pdf: 1349169 bytes, checksum: 611014d0a9479492b01703484a65b03f (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | 謝辭 I
摘要 II Abstract III 目錄 V 圖目次 VIII 表目次 IX 第一章 前言 1 1.1 石斑魚之簡介與台灣養殖概況 1 1.2 虹彩病毒 (Iridovirus) 2 1.2.1 虹彩病毒之分類 2 1.2.2 虹彩病毒之特性 4 1.2.3 石斑魚虹彩病毒極早期基因 5 1.3 病毒極早期基因之功能 5 1.4 TNF-α 6 1.5 TNFR 7 1.6 細胞凋亡(Apoptosis) 7 1.6.1 細胞凋亡及細胞壞死 7 1.6.2 細胞凋亡路徑 8 1.7 研究動機 8 第二章 材料與方法 10 實驗材料與藥品試劑 10 實驗方法 15 2.1 細胞培養 (cell culture) 15 2.1.1 細胞株及培養條件 15 2.1.2 細胞繼代 (passage) 培養 15 2.2 表現載體構築 (construction of plasmids) 15 2.3 勝任細胞 (competent cell) 的製備 16 2.4 轉形作用 (Transformation) 17 2.5 細胞轉染 (Transfection) 17 2.6 大腸桿菌重組蛋白之表現 18 2.7 大腸桿菌重組蛋白之純化 18 2.8 SDS-PAGE 電泳 18 2.9 西方墨點法 (Western blot) 19 2.10 免疫螢光染色 20 第三章 結果 21 3.1 GIV030L的序列與特性之分析 21 3.2 點帶石斑魚 TNF-α 之序列分析 21 3.3 轉染GIV030L 無法阻擋紫外光及Actinomycin D 誘導HeLa細胞凋亡 23 3.4 點帶石斑魚 TNF-α 重組蛋白之表現與純化 22 3.5 以重組TNF-α 誘發 GK 及 HeLa 細胞凋亡 22 3.6 以不同濃度之重組TNF-α搭配 Actinomycin D 處理 GK 細胞之影響 23 第四章 討論 24 第五章 參考文獻 28 圖目次 圖一、 石斑魚虹彩病毒030L之基因與胺基酸序列 34 圖二、 GIV030L序列及其蛋白結構分析 35 圖三、 石斑魚虹彩病毒030L胺基酸序列穿膜區域預測 36 圖四、 石斑魚 TNF-α 之基因與胺基酸序列 37 圖五、 石斑魚 TNF-α 之蛋白質結構分析 38 圖六、 真核細胞表現載體 pcDNA3CF-030L 之建構 39 圖七、 表現030L蛋白的HeLa細胞由紫外光照射後 引發HeLa細胞凋亡 40 圖八、 表現 030L 蛋白的 HeLa 細胞處理 Actinomycin D 後引發 HeLa 細胞凋亡 41 圖九、 原核系統表現載體 pET-43.1a(+)-Factor Xa-TNF-α 之建構 42 圖十、 利用四種不同大腸桿菌勝任細胞表現 TNF-α可溶性融合蛋白 43 圖十一、利用Ni2+親和性層析管柱純化 NusA-Factor Xa-TNF-α融合蛋白 44 圖十二、經由Factor Xa 酵素切割得到重組TNF-α 蛋白 45 圖十三、以重組TNF-α 處理 GK 及 HeLa 細胞 46 圖十四、相同濃度之Actinomycin D 與 不同劑量之重組TNF-α 造成 GK 細胞凋亡 47 圖十五、 石斑魚虹彩病毒029L及065R胺基酸序列穿膜區域預測 48 表目次 表一、 實驗使用之引子序列 49 | |
| dc.language.iso | zh-TW | |
| dc.subject | 石斑魚虹彩病毒 | zh_TW |
| dc.subject | 極早期基因 | zh_TW |
| dc.subject | GIV | en |
| dc.subject | iridovirus | en |
| dc.title | 石斑魚虹彩病毒極早期基因030L之分子特性表現及其功能之分析 | zh_TW |
| dc.title | Molecular characterization, expression and functional analysis of grouper iridovirus immediate-early gene 030L | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蘇建國(Jyan-Gwo J. Su),林正輝(Cheng-Hui Lin) | |
| dc.subject.keyword | 石斑魚虹彩病毒,極早期基因, | zh_TW |
| dc.subject.keyword | GIV,iridovirus, | en |
| dc.relation.page | 50 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2013-08-17 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 漁業科學研究所 | zh_TW |
| 顯示於系所單位： | 漁業科學研究所 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-102-1.pdf 未授權公開取用 | 1.32 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。