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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 蔡懷楨 | |
dc.contributor.author | Chia-Ling Kao | en |
dc.contributor.author | 高佳玲 | zh_TW |
dc.date.accessioned | 2021-06-08T04:48:16Z | - |
dc.date.copyright | 2009-07-30 | |
dc.date.issued | 2009 | |
dc.date.submitted | 2009-07-29 | |
dc.identifier.citation | 參考文獻
楊秉熹. (2008). Zebrafish as bioreactors to produce recombinant protein. 國立台灣大學分子與細胞生物學研究所碩士論文 Aguilera, O., Ostolaza, H., Quiros, L.M. and Fierro, J.F. Permeabilizing action of an antimicrobial lactoferricin-derived peptide on bacterial and artificial membranes. Febs Letters 462 (1999), pp. 273-277. Bellamy, W., Takase, M., Yamauchi, K., Wakabayashi, H., Kawase, K. and Tomita, M. Identification of the bactericidal domain of lactoferrin. Biochimica Et Biophysica Acta 1121 (1992), pp. 130-136. Britigan, B.E., Lewis, T.S., Waldschmidt, M., McCormick, M.L. and Krieg, A.M. Lactoferrin binds CpG-containing oligonucleotides and inhibits their immunostimulatory effects on human B cells. Journal of Immunology 167 (2001), pp. 2921-2928. Caipang, C.M.A., Verjan, N., Ooi, E.L., Kondo, H., Hirono, I., Aoki, T., Kiyono, H. and Yuki, Y. Enhanced survival of shrimp, Penaeus (Marsupenaeus) japonicus from white spot syndrome disease after oral administration of recombinant VP28 expressed in Brevibacillus brevis. Fish & Shellfish Immunology 25 (2008), pp. 315-320. Canada-Canada, F., Munoz de la Pena, A. and Espinosa-Mansilla, A. Analysis of antibiotics in fish samples. Analytical & Bioanalytical Chemistry (2009). Cerenius, L. and Soderhall, K. The prophenoloxidase-activating system in invertebrates. Immunological Review 198 (2004), pp. 116-26. Chapple, D.S., Hussain, R., Joannou, C.L., Hancock, R.E.W., Odell, E., Evans, R.W. and Siligardi, G. Structure and association of human lactoferrin peptides with Escherichia coli lipopolysaccharide. Antimicrobial Agents and Chemotherapy 48 (2004), pp. 2190-2198. Chen, L.L., Wang, H.C., Huang, C.J., Peng, S.E., Chen, Y.G., Lin, S.J., Chen, W.Y., Dai, C.F., Yu, H.T., Wang, C.H., Lo, C.F. and Kou, G.H. Transcriptional analysis of the DNA polymerase gene of shrimp white spot syndrome virus. Virology 301 (2002), pp. 136-147. Chou, H.Y., Huang, C.Y., Wang, C.H., Chiang, H.C. and Lo, C.F. Pathogenicity of a baculovirus infection causing white spot syndrome in cultured penaeid shrimp in Taiwan. Diseases of Aquatic Organisms 23 (1995), pp. 165-173. Demain, A.L. and Vaishnav, P. Production of recombinant proteins by microbes and higher organisms. Biotechnology Advances 27 (2009), pp. 297-306. Durand, S., Lightner, D.V., Nunan, L.M., Redman, R.M., Mari, J. and Bonami, J.R. Application of gene probes as diagnostic tools for White Spot Baculovirus (WSBV) of penaeid shrimp. Diseases of Aquatic Organisms 27 (1996), pp. 59-66. Fadok, V.A., de Cathelineau, A., Daleke, D.L., Henson, P.M. and Bratton, D.L. Loss of phospholipid asymmetry and surface exposure of phosphatidylserine is required for phagocytosis of apoptotic cells by macrophages and fibroblasts. Journal of Biological Chemistry 276 (2001), pp. 1071-1077. Fischer, R., Liao, Y.C., Hoffmann, K., Schillberg, S. and Emans, N. Molecular farming of recombinant antibodies in plants. Biological Chemistry 380 (1999), pp. 825-839. Fu, L.L., Li, W.F., Du, H.H., Dai, W. and Xu, Z.R. Oral vaccination with envelope protein VP28 against white spot syndrome virus in Procambarus clarkii using Bacillus subtilis as delivery vehicles. Letters in Applied Microbiology 46 (2008), pp. 581-586. Hansen, J.E., Lund, O., Tolstrup, N., Gooley, A.A., Williams, K.L. and Brunak, S. NetOglyc: Prediction of mucin type O-glycosylation sites based on sequence context and surface accessibility. Glycoconjugate Journal 15 (1998), pp. 115-130. Inouye, K., Miwa, S., Oseko, N., Nakano, H., Kimura, T., Momoyama, K. and Hiraoka, M. Mass mortalities of cultured kuruma shrimp penaeus-japonicus in japan in 1993 - electron-microscopic evidence of the causative virus. Fish Pathology 29 (1994), pp. 149-158. Jenssen, H., Andersen, J.H., Uhlin-Hansen, L., Gutteberg, T.J. and Rekdal, O. Anti-HSV activity of lactoferricin analogues is only partly related to their affinity for heparan sulfate. Antiviral Research 61 (2004), pp. 101-109. Johansson, M.W., Holmblad, T., Thornqvist, P.O., Cammarata, M., Parrinello, N. and Soderhall, K. A cell-surface superoxide dismutase is a binding protein for peroxinectin, a cell-adhesive peroxidase in crayfish. Journal of Cell Science 112 (1999), pp. 917-25. Johnson, K.N., van Hulten, M.C.W. and Barnes, A.C. 'Vaccination' of shrimp against viral pathogens: Phenomenology and underlying mechanisms. Vaccine 26 (2008), pp. 4885-4892. Kobayashi, M., Johansson, M.W. and Soderhall, K. The 76 kD cell-adhesion factor from crayfish hemocytes promotes encapsulation in vitro. Cell and Tissue Research 260 (1990), pp. 13-18. Kullberg, B.J., Netea, M.G., Vonk, A.G. and van der Meer, J.W.M. Modulation of neutrophil function in host defense against disseminated Candida albicans infection in mice. Fems Immunology and Medical Microbiology 26 (1999), pp. 299-307. Kumar, S.R., Ahamed, V.P.I., Sarathi, M., Basha, A.N. and Hameed, A.S.S. Immunological responses of Penaeus monodon to DNA vaccine and its efficacy to protect shrimp against white spot syndrome virus (WSSV). Fish & Shellfish Immunology 24 (2008), pp. 467-478. Kurtz, J. and Franz, K. Evidence for memory in invertebrate immunity. Nature 425 (2003), pp. 37-38. Li, L., Lin, S.M. and Yanga, F. Functional identification of the non-specific nuclease from white spot syndrome virus. Virology 337 (2005), pp. 399-406. Li, S.-S. and Tsai, H.-J. Transgenic microalgae as a non-antibiotic bactericide producer to defend against bacterial pathogen infection in the fish digestive tract. Fish & Shellfish Immunology 26 (2009), pp. 316-325. Lu, Y.N., Liu, J.J., Jin, L.J., Li, X.Y., Zhen, Y.H., Xue, H.Y., You, J.S. and Xu, Y.P. Passive protection of shrimp against white spot syndrome virus (WSSV) using specific antibody from egg yolk of chickens immunized with inactivated virus or a WSSV-DNA vaccine. Fish & Shellfish Immunology 25 (2008), pp. 604-610. Lupetti, A., Paulusma-Annema, A., Welling, M.M., Senesi, S., Van Dissel, J.T. and Nibbering, P.H. Candidacidal activities of human lactoferrin peptides derived from the N terminus. Antimicrobial Agents and Chemotherapy 44 (2000), pp. 3257-3263. Mader, J.S., Smyth, D., Marshall, J. and Hoskin, D.W. Bovine lactoferricin inhibits basic fibroblast growth factor- and vascular endothelial growth factor 165-induced angiogenesis by competing for heparin-like binding sites on endothelial cells. The American Journal of Pathology 169 (2006), pp. 1753-66. Masson, P.L., Heremans, J.F. and Dive, C. An iron-binding protein common to many external secretions. Journal of Clinical Chemistry 14 (1966), pp. 735-739. Murtha, J.M. and Keller, E.T. Characterization of the heat shock response in mature zebrafish (Danio rerio). Experimental Gerontology 38 (2003), pp. 683-691. Park, J.H., Lee, Y.S., Lee, S. and Lee, Y. An infectious viral disease of penaeid shrimp newly found in Korea. Diseases of Aquatic Organisms 34 (1998), pp. 71-75. Rout, N., Kumar, S., Jaganmohan, S. and Murugan, V. DNA vaccines encoding viral envelope proteins confer protective immunity against WSSV in black tiger shrimp. Vaccine 25 (2007), pp. 2778-2786. Santoro, M.G. Heat shock factors and the control of the stress response. Biochemical Pharmacology 59 (2000), pp. 55-63. Sarathi, M., Simon, M.C., Venkatesan, C. and Hameed, A.S.S. Oral administration of bacterially expressed VP28dsRNA to protect Penaeus monodon from white spot syndrome virus. Marine Biotechnology 10 (2008), pp. 242-249. Soderhall, K. and Smith, V.J. Separation of the haemocyte populations of Carcinus maenas and other marine decapods, and prophenoloxidase distribution. Development & Comparative Immunology 7 (1983), pp. 229-39. Soderhall, K., Wingren, A., Johansson, M.W. and Bertheussen, K. The cytotoxic reaction of hemocytes from the freshwater crayfish, Astacus astacus. Cellular Immunology 94 (1985), pp. 326-32. Sritunyalucksana, K., Wannapapho, W., Lo, C.F. and Flegel, T.W. PmRab7 is a VP28-binding protein involved in white spot syndrome virus infection in shrimp. Journal of Virology 80 (2006), pp. 10734-10742. Swartz, J.R. Advances in Escherichia coli production of therapeutic proteins. Current Opinion in Biotechnology 12 (2001), pp. 195-201. Thornqvist, P.O., Johansson, M.W. and Soderhall, K. Opsonic activity of cell adhesion proteins and beta-1,3-glucan binding proteins from two crustaceans. Developmental & Comparative Immunology 18 (1994), pp. 3-12. Tomita, M., Bellamy, W., Takase, M., Yamauchi, K., Wakabayashi, H. and Kawase, K. Potent antibacterial peptides generated by pepsin digestion of bovine lactoferrin. Journal of Dairy Science 74 (1991), pp. 4137-4142. Tomiya, N., Narang, S., Lee, Y.C. and Betenbaugh, M.J. Comparing N-glycan processing in mammalian cell lines to native and engineered lepidopteran insect cell lines. Glycoconjugate Journal 21 (2004), pp. 343-360. Tsai, J.M., Wang, H.C., Leu, J.H., Hsiao, H.H., Wang, A.H.J., Kou, G.H. and Lo, C.F. Genomic and proteomic analysis of thirty-nine structural proteins of shrimp white spot syndrome virus. Journal of Virology 78 (2004), pp. 11360-11370. Ueta, E., Tanida, T. and Osaki, T. A novel bovine lactoferrin peptide, FKCRRWQWRM, suppresses Candida cell growth and activates neutrophils. Journal of Peptide Reserch 57 (2001), pp. 240-9. van Hulten, M.C.W., Goldbach, R.W. and Vlak, J.M. Three functionally diverged major structural proteins of white spot syndrome virus evolved by gene duplication. Journal of General Virology 81 (2000a), pp. 2525-2529. van Hulten, M.C.W., Westenberg, M., Goodall, S.D. and Vlak, J.M. Identification of two major virion protein genes of White Spot Syndrome virus of shrimp. Virology 266 (2000b), pp. 227-236. van Hulten, M.C.W., Reijns, M., Vermeesch, A.M.G., Zandbergen, F. and Vlak, J.M. Identification of VP19 and VP15 of white spot syndrome virus (WSSV) and glycosylation status of the WSSV major structural proteins. Journal of General Virology 83 (2002), pp. 257-265. van Hulten, M.C.W. and Vlak, J.M. Identification and phylogeny of a protein kinase gene of white spot syndrome virus. Virus Genes 22 (2001), pp. 201-207. Wang, Y.G., Hassan, M.D., Shariff, M., Zamri, S.M. and Chen, X. Histopathology and cytopathology of white spot syndrome virus (WSSV) in cultured Penaeus monodon from peninsular Malaysia with emphasis on pathogenesis and the mechanism of white spot formation. Diseases of Aquatic Organisms 39 (1999), pp. 1-11. Witteveldt, J., Van Hulten, M.C.W. and Vlak, J.M. Identification and phylogeny of a non-specific endonuclease gene of white spot syndrome virus of shrimp. Virus Genes 23 (2001), pp. 331-337. Witteveldt, J., Vlak, J.M. and van Hulten, M.C.W. Protection of Penaeus monodon against white spot syndrome virus using a WSSV subunit vaccine. Fish & Shellfish Immunology 16 (2004), pp. 571-579. Yang, F., He, J., Lin, X.H., Li, Q., Pan, D., Zhang, X.B. and Xu, X. Complete genome sequence of the shrimp white spot bacilliform virus. Journal of Virology 75 (2001), pp. 11811-11820. Yi, G.H., Wang, Z.M., Qi, Y.P., Yao, L.G., Qian, J. and Hu, L.B. Vp28 of shrimp white spot syndrome virus is involved in the attachment and penetration into shrimp cells. Journal of Biochemistry and Molecular Biology 37 (2004), pp. 726-734. Zhan, W.B., Wang, Y.H., Fryer, J.L., Yu, K.K., Fukuda, H. and Meng, Q.X. White spot syndrome virus infection of cultured shrimp in China. Journal of Aquatic Animal Health 10 (1998), pp. 405-410. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23222 | - |
dc.description.abstract | 摘要
本研究欲以斑馬魚作為生物反應器大量表現外源抗病蛋白,分別建立斑馬魚表現抗菌蛋白lactoferricin (LFB)及抗白點病毒 (white spot syndrome virus, WSSV)的膜鞘蛋白VP28兩種系統。先前楊 (2008)建立以β-actin promoter驅動LFB-GFP持續表現之轉殖品系ZBL-5。本實驗取有綠螢光表現的ZBL-5之F2胚胎經synthetic gastric acid作用使LFB釋出,結果發現具有殺E. tarda的效果,並且一顆來自ZBL-5品系72 hpf之胚胎所釋出的LFB相當於2.88 ng ampicillin的效力。而利用ELISA進行定量,可知平均一顆72 hpf的F2胚胎約含有0.355 ng的LFB-GFP。此外,餵50顆3-dpf ZBL-5的F2胚胎給斑馬魚成魚,之後浸泡在1 × 105 CFU/ml的E. tarda菌液中,七天後平均存活率約75± 12.5%,而相同條件下餵食wild type胚胎的斑馬魚其存活率僅約4 ± 7.2%。因此證明ZBL-5轉殖品系所表現的LFB-GFP經餵食後在魚的消化道中可釋出具有抵抗病原菌E. tarda感染功效的LFB。此外,本實驗也建立能表現WSSV膜鞘蛋白VP28的斑馬魚品系,以使應用於草蝦口服疫苗。首先構築兩種基因轉殖質體:分別利用β-actin promoter驅動VP28-GFP表現之pBVPG質體及經熱誘導後以zebrafish heat shock promoter 70/4驅動VP28-GFP之pHVPG質體。注射pBVPG質體到約1500顆的斑馬魚胚胎中,挑選其中100顆有綠螢光均勻分布表現的胚胎,經遺傳配對後獲得4個品系;注射pHVPG質體到約3000顆斑馬魚胚胎中,挑選經熱休克後其中300顆有綠螢光均勻表現的胚胎,經遺傳配對後獲得7個品系。根據綠螢光的表現強度,選用HVPG-16之品系進行後續實驗。HVPG-16 之F2子代胚胎經過熱誘導後約有47 % ( 211/225, 249/291)有綠螢光表現,抽取這些胚胎之genomic DNA,利用PCR方法,可偵測到約1.4 kb的 VP28-GFP之外源基因片段;並利用Western blot方法,以 GFP抗體進行偵測2個月大成魚的全蛋白,發現有60 kDa及70 kDa訊號出現,而60 kDa的訊號大小與VP28-GFP蛋白片段分子量相符合。因此,本研究證實斑馬魚可以穩定表現外源病毒膜鞘蛋白。未來進行使蝦子口服抗病毒蛋白之實驗以預期應用於水產養殖以減少疾病所造成的損失。 | zh_TW |
dc.description.abstract | Abstract
In this study, we used zebrafish as a bioreactor to produce the functional proteins that enabled fish to resist the infection of pathogens. There were two proteins we developed: one was lactoferricin (LFB), an antimicrobial peptide; another was the envelope protein VP28 of white spot syndrome virus (WSSV), a WSSV-resistant protein. A LFB-harboring transgenic line generated by Yang (2008), ZBL-5, expressed LFB-GFP driven by β-actin promoter. We proved that the extracts isolated from the GFP-positive embryos derived from F2 generation of ZBL-5 displayed the inhibition ability against Edwardsilla tarda. After pepsin digestion, a 72-hpf embryo from ZBL-5 produced about 0.355 ng of exogenous LFB-GFP, and its bactericidal efficacy was equivalent to that of 2.88 ng ampicillin. When we fed adult zebrafish with 50 embryos of ZBL-5 and then immersed in water containing 1×105 CFU/ml E. tarda for 7 days, the survival rate was as high as 75±12.5%. However, the survival rate of zebrafish which fed with 50 wild-type embryos and challenged with E. tarda at same condition was only 4 ± 7.2%. These results clearly proved that the LFB-GFP produced by transgenic line ZBL-5 exhibited an antimicrobial LFB domain to kill a freshwater pathogen. Meanwhile, we also generated zebrafish transgenic lines which produced VP28 to serve as oral vaccine against WSSV for P. monodon. Firstly, we constructed two expression plasmids, in which VP28-GFP was driven by either β-actin promoter (pBVPG) or heat-inducible zebrafish heat shock promoter 70/4 (pHVPG). After the NotI-cut pBVPG was microinjected into 1500 one-celled zebrafish embryos, selected 100 GFP-positive G0 transgenic founders, and crossed with wild-type individually. Four G0 lines which produced GFP-positive F1 offspring were generated. Similarly, the NotI-cut pHVPG was microinjected into 2000 one-celled embryos, selected 300 GFP-positive G0 transgenic founders, and crossed with wild-type individually. There were seven G0 lines which produced GFP-positive F1 offspring were generated. According to the appearance of GFP intensity, we selected a transgenic line, HVPG-16, for further study due to its GFP was highly expressed after heat shock induction. We extracted genomic DNA of F2 embryos from HVPG-16 and detected by PCR. A PCR-produce with molecular mass of 1417-bp was amplified, which was corresponding that of PCR product amplified from transgene fragment VP28-GFP. Furthermore, when the total proteins extracted from F2 2-month-old transgenic fish were subjected to western blot analysis using antiserum against GFP, two positive bands with 60 kDa and 70 kDa were found on the gel. The 60-kDa signal was corresponding to that of recombinant VP28-GFP protein. Interestingly, the GFP expression rate of the F2 transgenic embryos examined was 47% ( 211/225, 249/291), indicating that the single insertion of transgene in this stable heterozygotic transgenic line HVPG-16. Hence, in this study, we generated a transgenic zebrafish that produces the virus envelop protein VP28. The further study on the resistance of WSSV infection of tiger shrimp after feeding the feed powder mixed with VP28-containing embryos is in progress. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T04:48:16Z (GMT). No. of bitstreams: 1 ntu-98-R96b43011-1.pdf: 2674443 bytes, checksum: a2425735146c067888e13f123f6f96ad (MD5) Previous issue date: 2009 | en |
dc.description.tableofcontents | 目錄
中文摘要………………………………………………………………………… 1 英文摘要………………………………………………………………………… 2 動機……………………………………………………………………………….4 前言 第一部分: 斑馬魚表現lactoferricin…………………….……………………6 第二部分: 斑馬魚表現WSSV膜鞘蛋白VP28……..………………………..9 材料與方法………………………………………………………………………15 結果 第一部分: 斑馬魚表現lactoferricin…………………….…………..……….26 第二部分: 斑馬魚表現WSSV膜鞘蛋白VP28…………………..………….28 討論 第一部分: 斑馬魚表現lactoferricin…………………...……………….……32 第二部分: 斑馬魚表現WSSV膜鞘蛋白VP28…………………………...…34 參考文獻………………………………………………………………………....39 圖表……………………………………………………………………………....46 附錄………………………………………………………………………………54 | |
dc.language.iso | zh-TW | |
dc.title | 利用斑馬魚作為生物反應器表現抗病蛋白 | zh_TW |
dc.title | Zebrafish as a bioreactor to produce recombinant pathogen-resistant proteins | en |
dc.type | Thesis | |
dc.date.schoolyear | 97-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 嚴宏洋,鄭登貴,蕭世民 | |
dc.subject.keyword | 斑馬魚,白點病毒,牛乳鐵蛋白, | zh_TW |
dc.subject.keyword | zebrafish,WSSV,lactoferricin, | en |
dc.relation.page | 58 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2009-07-29 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
顯示於系所單位: | 分子與細胞生物學研究所 |
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