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研究生:陳立涵
研究生(外文):Li-Han Chen
論文名稱:魚類結病毒與魚類反轉錄病毒共感染寄主期間之交互作用研究
論文名稱(外文):The Interactions between Fish Nodavirus and Fish Retrovirus during Co-infection
指導教授:齊肖琪齊肖琪引用關係
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:動物學研究研究所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:55
中文關鍵詞:交互作用魚類結病毒魚類反轉錄病毒
外文關鍵詞:NNVSnRVvirus interactions
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SSN-1細胞株常被作為魚類神經壞死症病毒(NNV)之研究工具,但有魚類反轉錄病毒SnRV的持續性感染。Lee 等 (2002)將 SSN-1細胞上清液感染另一株對NNV有感受性的細胞株GF-1,成為SnRV持續性感染的SGF-1細胞,以便發現SnRV與NNV共感染期間一些交互作用的現象。本實驗目的在於詳細探討共感染期間SnRV與NNV的交互影響。實驗結果顯示,SnRV的反轉錄酶(RT)會將NNV的兩段單股正意RNA全長以高相似度反轉錄為cDNA,產生了NNV生活史中一個新的階段,並在兩段NNV的核酸序列上找到與SnRV RT辨識結合序列有40至50%相似度的一段序列,但相似序列皆於NNV RNA序列的中段,因此真正的反轉錄起始位置,還需進一步求證。另外,觀察NNV感染SGF-1期間的日變化,發現(1)NNV cDNA在感染後第三天才能測到,以real time PCR分析,發現此cDNA的量隨感染後天數增加而增加;(2)SnRV的RNA則在第四天就完全測不到,而且SnRVRNA量會隨感染後天數增加而減少;(3)NNV感染SGF-1上清液中的NNV以GF-1與 SGF-1兩種細胞定力價,發現第三天到第五天兩系統測出之力價有104 TCID50/ ml的差異。從日變化的結果推測,病毒力價的差異可能與細胞上SnRV的接受器是是否開放有關。此外,發現NNV與SnRV共感染後是否會有NNV cDNA的產生,取決於寄主細胞的種類。最後,人工感染實驗中結果,確定SnRV可以感染石斑魚苗,但自然界中尚沒有發現NNV與SnRV有共感染的現象。
SSN-1 cell line was applied in many research works of nervous necrosis virus (NNV), but it was SnRV-persistently infected. Lee et al. (2002) developed another SnRV-persistently infected cell line SGF-1 by inoculation of the culture supernatant of SSN-1 cells into GF-1 cells. The goal of this study is to observe the interaction of SnRV and NNV during co-infection of SGF-1 cells. Both NNV genomic RNAs were reversely transcribed into cDNA with high conservation and a new stage of NNV in the life cycle was created. The sequences which were 40% to 50% identity with the sequence of primer binding site (PBS) of SnRV reverse transcriptase (RT) were found in NNV genomic RNAs. However, the both sequences were in the middle site of NNV genomic RNAs. Therefore, further experiments will be necessary to find the real PBS of SnRV RT. During the co-infection in NNV-infected SGF-1 cells, NNV cDNA was detected since day 3, and the amount of NNV cDNA increased daily. In the other hand, the SnRV RNA was not detected since day 4, and the amount of SnRV RNA decreased day by day. The titers of the supernatants collected from NNV-infected SGF-1 cells in GF-1 cells, were obviously higher than that in SGF-1 cells and determined in day 3 to day 5. It was suggested that the change of the titer might be linked up the exciting of SnRV receptor. Besides, if NNV RNA was reversely transcribed, that depended on what the host was. Finally, the data showed NNV and SnRV could co-infected in vivo when artificial infected. But there was not any co-infection in nature been found.
目錄…………………………………………………………………………….I
圖表目次……………………………………………………………………..IV
摘要……………………………………………………………………………V
Abstract………………………………………………………………………VI
第一章 前言 …………………………………………….………………….....1
1. 病毒性神經壞死症…………………………….………………….….1
1.1 病毒性神經壞死症的發現與地理分布….………………….....1
1.2 病毒性神經壞死症的病徵 ……………….…………………....2
1.3 流行病學(Epidemiology)之研究………………….……….…2
2. 神經壞死症病毒……………………………………………………...3
2.1 神經壞死症病毒的分類與特性………………………………..3
2.2 神經壞死症病毒於宿主體內之分佈…………………..………4
2.3 診斷方法……………………………………………….….……6
3. 鱧魚反轉錄病毒……………………………………………..……….7
4. 兩種病毒共同感染交互作用…………………………………….…..7
第二章 材料與方法……………………………………………………….….9
1. 細胞株與病毒來源……………………………………………….….9
2. SnRV感染GF-1細胞株………………………………………….…9
3. RNA之萃取………………………………………………………….9
4. DNA之萃取………………………………………………………...10
5. 反轉錄-聚合酶連鎖反應(Reverse transcription-polymerase
chain reaction, RT-PCR)……………………………………..…..11
6. 聚合酶連鎖反應(Polymerase chain reaction, PCR)…………...13
7. NNV感染SGF-1細胞株…………………………………………..14
8. Inverse PCR………………………………………………………..15
9. DNA定序…………………………………………………………...16
10. 核酸序列比對…………………………………………………….16
11. NNV感染SGF-1之病毒力價日變化…………………………..…16
12. Nested PCR…………………………………………………….….16
13. Realtime PCR…………………………………………………..….17
14. ScBB的製備…………………………………………………….….18
15. NNV感染ScBB、SSN-1、SGF-1細胞株……………………….…18
16. SnRV感染BB細胞株……………………………………………...18
17. 養殖魚採樣………………………………………………………....19
18. 活體感染及採樣…………………………………………………....19
第三章 結果………………………………………………………………....20
1. SnRV可於SGF-1中完成其生活史……………………………....20
2. NNV感染SGF-1中含有NNV RNA2之cDNA片段……………..20
3. NNV感染SGF-1後會有NNV RNA1的cDNA產生……………..21
4. 以GCG系統找到NNV RNA上與SnRV RT辨認位相似的區….21
5. NNV感染SGF-1後第三天才得以PCR測到NNV的cDNA…….22
6. SnRV在SGF-1中之RNA量會因NNV的共感染而下降………..22
7. NNV感染SGF-1上清液之病毒力價在GF-1及SGF-1的日變有差異…………………………………………………………...........22
8. 共感染產生之影響會因細胞株不同而有差異…………………....23
9. 台灣養殖魚未發現NNV與SnRV共感染情形……………….…..23
10. 人工攻毒可使部分石斑魚苗帶原SnRV………………………….24
第四章 討論………………………………………………………………....25
1. SnRV RT反轉錄NNV RNA的可能機制………………………....25
2. SnRV RNA在NNV感染SGF-1過程中減少的原因……………..26
3. 造成NNV感染SGF-1後日變化之原因…………………………..26
4. NNV與SnRV共感染不同物種細胞時產生的差異……………....28
5. NNV與SnRV在活體中共感染之可能性…………………………29
6. 病毒間共感染未來研究方向之展望……………………………....30
第五章 參考文獻………………………………………………………………...31
Aiyar, A., D. Cobrinik, Z. Ge, H.-J. Kung, and J. Leis (1992) Interaction between U5 viral RNA and the TΨC loop of the tRNATrp primer are required for efficient initiation of reverse transcriptase. Journal of Virology 66, 2464-2472.
Alonso, M., RodrÍguez S, and Pérez-Prieto S.I.(2003)Virulence of Infectious hematopoietic necrosis virus and Infectious pancreatic necrosis virus coinfection in rainbow trout (Oncorhynchus mykiss) and nucleotide sequence analysis of the IHNV glycoprotein gene. Archives of Virology 148, 1507–1521.
Arimoto, M., Mushiake, K., Mizuta, Y., Nakai, T., Muroga, K., and Furusawa, I.(1992)Detection of striped jack nervous necrosis virus(SJNNV)by enzyme-linked immunosorbent assay(ELISA). Fish Pathology 27, 191-195.
Arimoto, M., Mori, K., Nakai, T., Muroga, K. and Furusuwa, I.(1993)Pathogenecity in the causative agent of viral nervous necrosis disease of striped jack, Pseudocarnax dentex(Bloch&Schneider). Journal of Fish Diseases 16, 461-469.
Arimoto, M., Sato, J., Maruyama, K., Mimura, G. and Furusuwa, I.(1996)Effect of chemical and physical treatments on the inactivation of striped jack nervous necrosis virus(SJNNV). Aquaculture 143, 15-22.
Bloch, B., Gravningen, K., and Larsen, J.L.(1991)Encephalomyelitis among turbot associated with a picornavirus-like agent. Diseases of Aquatic Organisms 10, 65-70.
Bovo, G., Maltese, C., Mutinelli, F., Rosin, R. and Montesi, F.(1999a)A rapid and efficient method for diagnosing viral encephalopathy in symptomatic specimens. Abstract P-162.: Ninth International Conference of the European Association of Fish Pathologistis, 19-24 September, Rhodes.
Bovo, G., Nishizawa, T., Maltese, C., Borghesan, F., Mutinelli, F., Montesi, F., and De Mas, S.(1999b)Viral encephalopathy and retinopathy of farmed marine fish species in Italy. Virus Research 63, 143-146.
Breuil, G., Bonami, J.R., Pepin, J.F., Pichot, Y.(1991)Viral infection(picona-like virus)asscociated with mass mortalities in hatchery-reared sea-bass(Dicentrarchus labrax)larvae and juveniles. Aquacultrue 97, 109-116.
Chang, S.F., Ngoh, G.H., Kueh, L.F.S., Qin, Q.W., Chen ,C.L., Lam, T.J. and Sin, Y.M.(2001)Development of a tropical marine fish cell line from Asian seabass(Lates calcarifer)for virus isolation. Aquaculture 192, 133-145.
Cheng, C.P., and Tsai, C.H.(1999)Structural and functional analysis of the 3’ untranslated region of bamboo mosaic potexvirus genomic RNA. Journal of Molecular Biology 288, 555-565.
Chi, S.C., Hu, W.W. and Lo, B.J.(1999)Establishment and characterization of a continuous cell line(GF-1)derived from grouper, Epinephelus coioides(Hamilton):a cell line susceptible to grouper nervous necrosis virus(GNNV). Journal of Fish Diseases 22, 1-10.
Chi, S.C., Lo, B.J. and Lin, S.C., Wen, W.W., Lo, C.F., Kou, G.H. & Chen, S.N.(2000)The survey of viral nervous necrosis among cultured groupers in Taiwan. In: APEC FWG02/2000. Bangkok, 18-20.
Chi, S.C., Lo, B.J. and Lin, S.C.(2001)Characterization of grouper nervous necrosis virus(GNNV). Journal of Fish Diseases 24, 3-13.
Chi, S.C., shieh, J.R., and Lin, S.J.(2003)enetic and antigenic analysis of betanodavirus isolated from aquatic organisms in Taiwan. Disease of Aquatic Organisms 55, 221-228.
Chiu, W.W., Hsu, Y.H., and Tsai, C.H.(2002)Specificity analysis of the conserved hexanucleotides for the replication of bamboo mosaic potexvirus RNA. Virus Research 83, 159-167.
Chomczynski, P. and Scchi, N(1987)Single-step method for RNA isolation by acid guanidium thiocyanate-phenol-chloroform extration. Analytical Biochemistry 162, 156-159.
Comps, M., Trindade, M., and Delsert, C.L.(1996)Investigation of fish encephalitis viruses(FEV)expression in marine fishes using DIG-labelled probes. Aquaculture 143, 113-121.
Dalla Valle, L., Zanella, L., Patarnello, P., Paolucci, L., Belvedere, P. and Colombo, L.(2000)Development of a sensitive diagnostic assay for fish nervous necrosis virus based on RT-PCR plus nested PCR. Journal of Fish Diseases 23, 321-327.
Dasgupta, R., Ghosh, A., Dasmahapatra, B., Guarino, L.A. and Kaesberg, P.(1984)Primary and secondary structure of black beetle virus RNA2, the genomic messenger for BBV coat protein. Nucleic Acid Research 12, 7215-7223.
Dasgupta, R. and Sgro, J.Y.(1989)Nucleotide sequences of three nodavirus RNA2’s : the messengers for their coat protein precursors. Nucleic Acid Research 17, 7525-7526.
Frerichs, G.N., Morgan, D., Hart, D., Skerrow, C., Roberts, R.J. and Onions, D.E.(1991)Spontaneously productive C-type retrovirus infection of fish cell lines. Journal of General Virology 72, 2537-2539.
Frerichs, G.N., Rodger, H.D., and Peric, Z.(1996)Cell culture isolation of piscine neuropathy nodavirus from juvenile sea bass , Dicentrchus labrax. Journal of Virology 77, 2067-2071.
Frerichs, G.N., Tweedie, A., Starkey, W.G., Richards, R.H.(2000)Temperature, pH and electrolyte sensitivity, and heat, UV and disinfectant inactivation of sea bass(Dicentrarchus labrax)neuropathy nodavirus. Aquaculture 185, 13-24.
Fondong, V. N., Pita, J. S., Rey, M. E., de Kochko, A., Beachy, R. N., and Fauquet, C. M. (2000) Evidence of synergism between African cassava mosaic virus and a new double-recombinant geminivirus infecting cassava in Cameroon. Journal of General Virology 81, 287-297.
Fukuda, Y., Neuyen, H.D., Furuhashi, M. and Nakai, T.(1996)Mass mortality of cultured sevenband grouper, Epinephelus septemfasciatus, associated with viral nervous necrosis. Fish Pathology 31, 165-170.
Glazebrook, J.S., Heaaman, M.P., and de Beer, S.W.(1990)Picona-like viral particles associated with mass mortalities in larval barramundi, lates calcarifer (Bloch). Journal of Fish Diseases 13, 245-249.
Grotmol, S., Totland, G.K., Kvellestad, A., Fjell, K., and Olsen, A.B.(1995)Mass mortality of larval and juveniles hatchery-reared halibut(Hippoglossus hippoglossus L.)associated with the presence of virus-like particles in vacuolated lesions in the central nervous system and retina. Bulletin of the European Association of Fish Pathologists 15, 176-180.
Grotmol, S., Totland, G.K., and Kryvi, H.(1997)Detection of a nodavirus-like agent in heart tissue from reared Atlantic salmon Salmo salar suffering from cardiac myopathy syndrome(CMS). Diseases of Aquatic Organisms 29, 79-84.
Grotmol, S., Bergh, O. and Totland, G.K.(1999)Transmission of viral encephalopathy and retinopathy(VER)to yolk-sac larvae of the Atlantic halibut Hippoglossus hippoglossus:occurrence of nodavirus in various organs and a possible route of infection. Diseases of Aquatic Organisms 39, 95-106.
Grotmol, S. and Totland, G.K.(2000)Surface disinfection of Atlantic halibut Hippoglossus hippoglossus eggs with ozonated sea-water inactivates nodavirus and increases survival of the larvae. Diseases of Aquatic Organisms 39, 89-96.
Guo, Y.X., Wei, T., Dallmann, K., and Kwang, J.(2003)Induction of caspase-dependent apoptosis by betanodavirus GGNNV and demonstration of protein α as an apoptosis inducer. Virology 308, 74-82.
Hart, D., Fereichs, G.N., Rambaut, A., and Onions, D.E.(1996) Complete nucleotide sequence and transcriptional analysis of the snakehead fish retrovirus. Journal of Virology 70, 3606-3616.
Hegde, A., The, H.C., Lam, T.J., and Sin, Y.M.(2003)Nodavirus infection in freshwater ornamental fish, guppy, Poicelia reticulate – comparative characterization and pathogenicity studies. Archives of Virology 148, 575-586.
Huang, C.Y., Huang, Y.L., Meng, M., Hsu, Y.H., and Tsai, C.H.(2001)Sequences at the 3’ untranslated region of bamboo mosaic potexvirus RNA interact with viral RNA-dependent RNA polymerase. Journal of Virology 75, 2818-2824.
Iwamoto, T., Mori, K., Arimoto, M. and Nakai, T.(1999)High permissivity of the fish cell line SSN-1 for piscine nodaviruses. Diseases of Aquatic Organisms 39, 37-47.
Iwamoto, T., Nakai, T., Mori, K., Arimoto, M. and Furusawa, I.(2000)Cloning of the fish cell line SSN-1 for piscine nodaviruses. Diseases of Aquatic Organisms 43, 81-89.
Iwamoto, T., Mori, K., Arimoto, M. and Nakai, T.(2001)A combined cell-culture and RT-PCR method for rapid detection of piscine nodaviruses. Journal of Fish Diseases 24, 231-236.
Jung, S.J., Miyazaki, T., Miyata, M., and Oishi, T.(1996)Histopathological studies on viral nervous necrosis in a new host Japanese sea bass Lateolabrax japonicus. Bulletin of the Faculty of Bioresources, Mie University 16, 9-16.
Kaesberg, P. and Johnson, J.E.(1990)Structural homology among four nodaviruses as dedused by sequencing and X-ray crystallography. Journal of Molecular Biology 214, 423-435.
Lai, Y.S., Chiu, H.C., Murali, S., Guo, I.C., Chen, S.C., Fang, K. and Chang, C.Y.(2001)In vitro neutralization by monoclonal antibodies against yellow grouper nervous necrosis virus (YGNNV)and immunolocalization of virus infection in yellow grouper, Epinephelus awoara(Temminck & Schlegel). Journal of Fish Diseases 24, 237-244.
Lai, Y.S., John, J.A.C., Guo, I.C., Chen, S.C., Fang K., Lin, C.H., and Chang, C.Y.(2003)Establishment of cell lines from a tropical grouper, Epinephelus awoara(Temminck & Schlegel), and their susceptibility to grouper irido- and nodaviruses. Journal of Fish Diseases 26, 31-42.
Le Breton, A., Grisez, L., Sweetman, J., and Ollevier, F.(1997)Viral nervous necrosis(VNN)associated with mass mortalities in cage-reared sea bass , Dicentrarchus labrax(L.). Journal of Fish Diseases 20, 145-151.
Lee, K. W., Chi, S.C., and Cheng, T. M. (2002) Interference of the life cycle of fish nodavirus with fish retrovirus. Journal of General Virology 83, 2469-2474.
Leis, J., Aiyar, A., and Cobrinik, D.(1993)Regulation of initiation of reverse transcription of retrovirus. Reverse Transcriptase, 33-45.
Lyles, D.S. (2000) Cytopathogenesis and inhibition of host gene expression by RNA viruses. Microbiology and Molecular Biology Review 64: 709-724.
Méndez-Lozano, J., Torres-Pacheco, I., Fauquet, C. M., and Rivera- Bustamante, R. F.(2003)Interactions between geminiviruses in a naturally occurring mixture: pepper huasteco virus and pepper golden mosaic virus. Phytopathology 93, 270-277.
Mori, K., Nakai, T., Negahara, M., Muroga, K., Mckuchi, T. and Kanno, T.(1991)A viral disease in hatchery-reared larvae and juveniles of redspotted grouper. Fish Pathology 26, 209-210.
Mori, K., Nakai, T., Muroga, K., Arimoto, M., Mushiake, K., and Furusawa, I.(1992)Properties of a new virus belonging to Nodaviridae found in larval striped jack(Pseudocaranx dentex)with nervous necrosis. Virology 187, 368-371.
Munday, B.L., Langdon, J.S., Hyatt, A., and Humphrey, J.D.(1992)Mass mortality associated with a viral-induced vacuolating encephalopathy and retinopathy of larval and juvenile barramundi, Lates calcarfer (Bloch). Aquaculture 103, 197-211.
Munday, B.L., Kwang, J., and Moody, N.(2002)Betanodavirus infection of teleost fish : a review. Journal of Fish Diseases 25, 127-142.
Muroga, K.(1995)Viral and bacterial disease in larval and juvenile marine fish and shellfish: a review. Fish Pathology 30, 71-85
Mushiake, K., Arimoto, M., Furusawa, T., Furusawa, I., Nakai, T. and Muroga, K.(1992)Detection of antibodies against striped jack nervous necrosis virus(SJNNV)from brood stocks of striped jack. Nippon Suisan Gakkaishi 58, 2351-2356.
Mushiake, K., Nishizawa, T., Nakai, T., Furusawa, I., and Muroga, K.(1994)Control of VNN in striped jack : selection of spawners based on the detection of SJNNV gene by polymerase chain reaction(PCR). Fish Pathology 29,177-182.
Nakai, T., Nguyen, H.D., Nishizawa, T., Muroga, K., Arimoto, M., and Ootsuki, K.(1994)Occurrence of viral nervous necrosis in kelp grouper and tiger puffer. Fish Pathology 29, 211-214.
Nguyen, H.D., Mekuchi, T., Imura, K., Nakai, T., Nishizawa, T., and Muroga, K.(1994)Occurrence of viral nervous necrosis(VNN)in hatchery-reared juvenile Japanese flounder Paralichthys olivaceus. Fish Science 60, 551-554.
Nguyen, H.D., Nakai, T., and Muroga, K.(1996)Progression of striped jack nervous necrosis virus(SJNNV) infection in naturally and experimentally infected striped jack Pseudocaranx dentex larvae. Diseases of Aquatic Organisms 24, 99-105.
Nishizawa, T., Mori, K., and Nakai, T.(1994)Polymerase chain reaction(PCR)amplification of RNA of striped jack nervous necrosis virus(SJNNV). Diseases of Aquatic Organisms 18, 103-107.
Nishizawa, T., Masaaki, K., Nakai, T. and Muroga, K.(1995)Comparison of the coat protein genes of five fish nodaviruses, the causative agent of nervous necrosis in marine fish. Journal of General Virology 76, 1563-1569.
Nishizawa, T., Furuhashi, M., Nagal, T. and Muroga, K.(1997)Genomic classification of fish nodaviruses by molecular phylogenetic analysis of the coat protein gene. Applied and Environmental Microbiology 63, 1633-1636.
Renault, T., Haffner, P.H., Baudin, L.F., Breuil, G., and Bonami, J.R.(1991)Mass mortalities in hatchery-reared sea bass(Lates calcarifer)larvae associated with the presence in the brain and retina of virus-like particles. Bulletin of the European Association of Fish Pathologists 11, 68-73.
Skliris, G.P. and Richards, R.H.(1998)Assessment of the susceptibility of the brine shrimp Artemia salina and rotifer Brachionus plicatilis to experimental nodavirus infection. Aquaculture 169, 133-141.
Telesnitsky, A. and Goff, S.P.(1993)Strong-stop strand transfer during reverse transcription. Reverse transcriptase, 49-83.
Van Regenmortel, M. H. V, Fauquet, C, Bishop, David H. L(2000) Family nodavirus. Virus taxonomy : classification and nomenclature of viruses : seventh report of the International Committee on Taxonomy of Viruses, 747-755.
Weiss, R., N. Teich, H. Varmus, and J. Coffin (1985) Molecular biology of tumor viruses, 2nd edition: RNA tumor viruses, parts 1 and 2. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Yoshikoshi, K., and Inoue, K.(1990)Viral nervous necrosis in hatchery- reared larvae and juveniles of Japanese parrotfish, Oplegnathus fasciatus(Temminck&Schlegel). Journal of Fish Diseases 13, 69-77.
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