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標題: 重組桿狀病毒在蟲害防治及外源蛋白表現的開發
Development of recombinant baculoviruses for insect pest control and foreign protein expression
作者: 靳子蓉
Jinn, Tzyy-Rong
關鍵字: Baculovirus
Insect pest control
Foreign protein expression
出版社: 生物科技學研究所
引用: Benkhalifa R, Stankiewicz M, Lapied B, Turkev M, Zilberberg N, Gurevitz M, Pelhate M (1997) Refined electrophysiological analysis suggests that a blocker. Life Sci 61:819-830 Carbonell LF, Hodge MR, Tomalski MD, Miller LK (1988) Synthesis of a gene coding for an insect-specific scorpion neurotoxin and attempts to express it using baculovirus vectors. Gene 73:409-418 Chang JH, Choi JY, Jin BR, Roh JY, Olszewski JA, Seo SJ, O'Reilly DR, Je YH (2003) An improved baculovirus insecticide producing occlusion bodies that contain Bacillus thuringiensis insect toxin. J. Invertebr. Pathol. 84:30-37 Chao YC, Lee ST, Chang MC, Chen HH, Chen SS, Wu TY, Liu FH, Hsu EL, Hou RF (1998) A 2.9-kilobase noncoding nuclear RNA functions in the establishment of persistent Hz-1 viral infection. J. Virol. 72:2233-2245 Chejanovsky N, Zilberberg N, Rivkin H, Zlotkin E, Gurevitz M (1995) Functional expression of an alpha anti-insect scorpion neurotoxin in insect cells and lepidopterous larvae. FEBS Lett. 376:181-184 Cory JS, Hails RS (1997) The ecology and biosafety of baculoviruses. Curr. Opin. Biotechnol. 8: 323-327 Cory JS, Hirst ML, Williams T, Hails RS, Goulson D, Green BM, Carty TM, Possee RD, Cayley PJ, Bishop DHL (1994) Field trial of a genetically improved baculovirus insecticide. Nature 370:138-140 Gershburg E, Stockholm D, Froy O, Rashi S, Gurevitz M, Chejanovsky N (1998) Baculovirus-mediated expression of a scorpion depressant toxin improves the insecticidal efficacy achieved with excitatory toxins. FEBS Lett. 422:132-136 Gordon D, Moskowitz H, Eitan M, Warner C, Catterall WA, Zlotkin E (1992) Localization of receptor sites for insect-selective toxins on sodium channels by site-directed antibodies. Biochemistry 31:7622-7628 Harrison RL, Bonning BC (2000) Use of scorpion neurotoxins to improve the insecticidal activity of Rachiplusia ou multicapsid nucleopolyhedrovirus. Biol. Control 17:191-201 Harrison RL, Bonning BC (2001) Use of proteases to improve the insecticidal activity of baculoviruses. Biol. Control 20: 199-209 Herrmann R, Moskowitz H, Zlotkin E, Hammock BD (1995) Positive cooperativity among insecticidal scorpion neurotoxins. Toxicon 33:1099-1102 Hung CC, Hwan JS (1988) The mass rearing method of major insect pests: Asian corn borer, Ostrinia furnacalis, guava mealy bug, Planococcus minor and beet armyworm, Spodoptera exigua. Annual Report TACTRI, pp.50-52, Tapei, Taiwan. Krapcho KJ, Kral RM, Van Wagenen BC, Eppler KJ, Morgan TK (1995) Characterization and cloning of insecticidal peptides from the primitive weaving spider Diguelia Canties. Insect Biochem. Mol. Biol. 25: 991-1000 Maeda S (1989) Increased insecticidal effect by a recombinant baculovirus carrying a synthetic diuretic hormone gene. Biochem. Biophys. Res. Commun. 165: 1177-1183 Maeda S (1995) Further development of recombinant baculovirus insecticides. Curr. Opin. Biotechnol. 6: 313-319 Maeda S, Volrath SL, Hanzlik TN, Harper SD, Majima K, Maddox DW, Hammock BD, Foeler E (1991) Insecticidal effects of an insect-specific neurotoxin expressed by a recombinant baculovirus. Virology 184: 777-780. Miller LK, Ling AJ, Bulla LA (1983) Bacterial, viral and fungal insecticides. Science 219: 715 -721 O'Reilly DR, Miller LK, Luckow VA (1992b) Baculovirus expression vector : a laboratory manual, W.H.Freeman and Company, New York. O'Reilly DR, Kelly TJ, Masler EP, Thyagaraja BS, Robson, RM (1993) Overexpression of Bombyx mori prot-horacicotrophic hormone using baculovirus vectors. Insect Biochem. Mol. Biol. 25: 475-485 Popham Holly JR, Prikhod'ko GG, Felcetto TJ, Ostlind DA, Warmke JW, Cohen CJ, Miller LK (1998) Effect of Deltamethrin treatment on lepidopteran larvae infected with baculoviruses expressing insect-selective toxins: u-aga-IV, AsII and ShI. Biol. Control 12: 79-87 Possee RD (1986) Cell-surface expression of influenza virus haemagglutinin in insect cells using a baculovirus vector. Virus Res. 543-559 Possee RD, Barnett AL, Hawtin RE, King LA (1999) Engineer baculoviruses for pest control. Pest Manag. Sci. 51: 462-470 Regev A, Rivkin H, Inceoglu B, Gershburg E, Hammock BD, Gurevitz M, Chejanovsky N (2003) Further enhancement of baculovirus insecticidal efficacy with scorpion toxins that interact cooperatively. FEBS Lett. 537: 106-110 Tuan SJ, Hou RJ, Kao SS, Lee CF, Chao YC (2005) Improved plant protective efficacy of a baculovirus using an early promoter to drive insect-specific neurotoxin expression. Bot. Bull. Acad. Sir. 46: 11-20 van Beek NAM, Hughes PR (2003) Effect of signal sequence and promoter on the speed of action of a genetically modified Autographa californica nucleopolyhedrovirus expressing the scorpion toxin LqhIT2. Biol. Control 27: 53-64 Zlotkin E, Gurevitz M, Fowler E, Adams Michael E (1993) Depressant insect selective neurotoxins from scorpion venom: chemistry, action, and gene cloning. Arch. Insect Biochem. Physiol. 22: 55-73
摘要: 重組桿狀病毒在蟲害防治及外源蛋白表現的開發 中文摘要 桿狀病毒(baculoviruses)已被成功的發展為生物農藥,其中以加州苜蓿夜蛾核多角體病毒(Autographa californica multiple nuclear polyhedrosis viruses ; AcMNPVs)最常被應用;但由於緩慢的殺蟲效果限制了桿狀病毒在農業害蟲的防治應用。此外,以安全評估的考量,如何追蹤重組桿狀病毒之基因在環境中的穩定性與存在亦是十分重要的議題。因此,vAcPhsp70EGFP/Ppag90IT2重組加州苜蓿夜蛾核多角體病毒被構築,使其同時表現綠螢光蛋白(EGFP)和以色列蠍毒殺蟲蛋白(LqhIT2),並分別以Phsp70及Ppag90啟動子啟動。結果顯示此重組病毒除能有效防治擬尺蠖幼蟲外,還可表現EGFP進行偵測已感染之病蟲。另一方面,由於珊瑚紅螢光蛋白(DsRed)能於接收太陽光後釋放出較長波長的光;因此在本論文研究中構築可同時表現EGFP 和DsRed的重組桿狀病毒vAcR-IR-G,以進一步探討EGFP 和DsRed二螢光蛋白在作為重組桿狀病毒(GMBV)追蹤劑的比較。結果顯示此重組病毒vAcR-IR-G在感染擬尺蠖、甜菜夜蛾和斜紋夜蛾幼蟲後,當其內含之DsRed蛋白表現,於日照下且不需任何其他輔助儀器,即可釋放紅螢光。因此,我們認為將此DsRed蛋白應用於偵測田間的重組桿狀病毒,將是一個有效的追蹤劑。自1983年起,桿狀病毒/昆蟲細胞表現系統即被廣泛的用來生產許多不同的生物醫療蛋白。故本研究以此系統來表現松杉靈芝(Ganoderma tsugae)中所分離出的免疫調節蛋白FIP-gts,結果依生物活性來評估,由Sf21昆蟲細胞所產生的rFIP-gts比由E.coli細胞所產生的rFIP-gts佳。
Development of Recombinant Baculoviruses for Insect Pest Control and Foreign protein expression Abstract Baculoviruses have been successfully developed as biological control agents, especially Autographa californica multiple nucleopolyhedrosis viruses (AcMNPVs), which is used most frequently. However, the slow effectiveness limits their application in agriculture. In a safety standpoint, it is also important to track the genetic stability of recombinant baculoviruses and their presence in the environments. Thus, a polyhedrin-positive recombinant AcMNPV vAcPhsp70EGFP/Ppag90IT2 was constructed for larvae to express the EGFP and scorpion neurotoxin LqhIT2 under the control of Phsp70 and Ppag90 promoter, respectively. These results revealed that recombinant AcMNPV vAcPhsp70EGFP/Ppag90IT2 may significantly increase the insecticidal potency against Trichoplusia ni when additional EGFP was expressed as a visible marker. On the other hand, the red-protein pigment of DsRed, a red fluorescence protein, may convert the short wavelength light component of the solar radi¬ation into a longer wavelength light. Thus, the EGFP and DsRed fluorescent genes are also expressed simultaneously in insect larva to evaluate which fluorescent protein is relatively bright and is suitable to act as GMBV tracer. Results demonstrate the insect larvae, including Tri¬choplusia ni, Spodoptera exigua, and Spodoptera litura, infected by the genetic modified baculovirus (GMBV) containing DsRed gene can emit red fluorescence under sun light without any prosthetic apparatus. So, DsRed may serve as a powerful tracer to evaluate the efficiency of GMBV as an insecticide in the field. Since 1983, the baculovirus-insect cell expression system has been widely used to produce recombinant proteins for many different biomedical applications. In this study, the baculovirus/insect cell expression system is utilized to produce active FIP-gts, a fungal immunomodulatory protein from Song-Shan Lingzhi (Ganoderma tsugae). As expected, rFIP-gts produced in Sf21 cells is a better source than that produced in E. coli cells for the application of oral administration.
其他識別: U0005-2807200610382000
Appears in Collections:生物科技學研究所



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