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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王重雄(Chung-Hsiung Wang) | |
| dc.contributor.author | Yi-Ting Yang | en |
| dc.contributor.author | 楊怡婷 | zh_TW |
| dc.date.accessioned | 2021-06-14T16:58:45Z | - |
| dc.date.available | 2008-08-05 | |
| dc.date.copyright | 2008-08-05 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-07-28 | |
| dc.identifier.citation | 林光宏。2006。木毒蛾核多角體病毒 (LyxyMNPV) 量產用之舞毒蛾細胞 (IPLB-LD-652Y) 次選殖株系篩選。碩士論文。國立台灣大學昆蟲學研究所。臺北。中華民國。
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Baculovirus vectors for expression of foreign genes. Adv Virus Res 35: 177-192. Lane, E. B. 1982. Monoclonal antibodies provide specific intramolecular markers for the study of epithelial tonofilament organization. J Cell Biol 92: 665-673. Lynn, D. E. 1989. Methods for the development of cell lines from insects. J Tissue Cult Meth 12: 23-29. Lynn, D. E. 1999. Development of insect cell lines: virus susceptibility and applicability to prawn cell culture. Methods Cell Sci 21: 173-181. Lynn, D. E. 2001. Novel techniques to establish new insect cell lines. In Vitro Cell Dev Biol Anim 37: 319-321. Lynn, D. E., Dougherty, E. M., McClintock, J. T., and Loeb, M. 1988. Development of cell lines from various tissues of lepidoptera. Invertebrate and fish tissue culture 1988: 239-242. Maeda, S. 1989. Expression of foreign genes in insects using baculovirus vectors. Annu Rev Entomol 34: 351-372. Miller, L. K. 1988. Baculoviruses for foreign gene expression in insect cells. Biotechnology 10: 457-465. Miller, L. K. 1997. The baculoviruses. Plenum Press, New York, 141-170. Moll, R., Franke, W. W., Schiller, D. L., Geiger, B., and Krepler, R. 1982. The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells. Cell 31: 11-24. Morris, T. D., and L. K. Miller. 1994. Mutational analysis of a baculovirus major late promoter. Gene 140: 147-153. Okano, K., A. L. Vanarsdall, V. S. Mikhailov, G. F. Rohrmann. 2006. Conserved molecular systems of the baculovirus. Virology 344: 77-87. Ooi, B. G., C. Rankin, and L. K. Miller. 1989. Downstream sequences augment transcription from the essential initiation site of a baculovirus polyhedrin gene. J Mol Biol 210: 721-736. Pijlman, G. P., E. van den Born, D. E. Martens, and J. M. Vlak. 2001. Autographa californica baculoviruses with large genomic deletions are rapidly generated in infected insect cells. Virology 283: 132-138. Pullen, S. S., and P. D. Friesen, 1995. The CAGT motif functions as an initiator element during early transcription of the baculovirus transregulator ie-1. J Virol 69: 3575-3583. Quiot, J. M. 1976. Establishment of a cell line ovaries. C. R. Acad. Sci. Ser. D 282: 465-467. Reed, L.J. and H. Muench. 1938. A simple method of estimating fifty percent endpoints. The American Journal of Hygiene 27: 493–497. Rothfels, K. H., and Siminovitch, L. 1958. An air-drying technique for flattening chromosomes in mammalian cells grown in vitro. Stain Technol 33: 73-77. Schafer, M. P., G. Rohrmann, U. Heine, and G. S. Beaudreau. 1979. DNA from two Orgyia pseudotsugata baculoviruses: Molecular weight determination by means of electron microscopy and restriction endonuclease analysis. Virology 95: 176-184. Schneider, I. 1969. Establishment of three diploid cell lines of Anopheles stephensi (Diptera: Culicidae). J Cell Biol 42: 603-606. Schneider, I. 1972. Cell lines derived from late embryonic stages of Drosophila melanogaster. 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The chemistry of insect hemolymph; organic components of the hemolymph of the silkworm, Bombyx mori, and two other species. J Gen Physiol 39: 853-868. Yeh, S. C., S. T. Lee, C. Y. Wu, and C. H. Wang. 2007. A cell line (NTU-MV) established from Maruca vitrata (Lepidoptera: Pyralidae): Characterization, viral susceptibility, and polyhedra production. J Invertebr Pathol 96: 138-146. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40751 | - |
| dc.description.abstract | 昆蟲體外細胞培養系統的發展，已將近一世紀，至今此技術廣泛的應用於桿狀病毒的研究以及重組蛋白的生產。吉普賽舞蛾 (Lymantria dispar) 細胞株IPLB-LD-652Y經本實驗選殖後，一共得到六個子細胞株IPLB-LD-652Y-a~f (簡稱LD-a ~ -f)。這些子細胞具有不同比例的細胞形態，包括圓形 (round-shaped)、紡錘形 (spindle-shaped)、鱗形 (squamous-shaped) 與多態形 (polymorphous-shaped)。 LD-a細胞以紡錘形細胞最多；其中 LD-d細胞具有雙臂很長的紡錘形細胞 (類感覺神經細胞)；LD-b、-c和-f則有較多圓形細胞。這些子細胞株與其母株具有相同的酯酶圖譜 (esterase pattern)。 LD-b與 -f的細胞質含有較多的脂肪顆粒，但兩者的分布狀況不同，LD-b脂肪顆粒均勻散布在細胞質中，而LD-f則呈聚集狀。所有的細胞子株皆對野生型黑角舞蛾核多角體病毒株 (Lymantria xylina multiple nucleopolyhedrovirus)，LyxyMNPV 和LyxyNPV-2具有高感受性，感染後14天之平均感染率可達98%。其中LD-a 感染黑角舞蛾核多角體病毒產生較高多角體產量，較低胞外病毒效價；反之，LD-d感染黑角舞蛾核多角體病毒為較少多角體產量與較低胞外病毒效價。另外，在螢光重組病毒LyxyExp-EGFP和LyxyExp-DsRed感染實驗中，LD-a產生最高紅、綠螢光蛋白產量；LD-d則最少。本實驗結果顯示子細胞株系之形態、特性以及病毒感受性上不同於母株，尤其是子細胞LD-a可產生較母株高之重組蛋白產量，具有生產重組蛋白的潛力。 | zh_TW |
| dc.description.abstract | Insect in vitro culture system has been developed for a century and wild applied in study of baculovirus and the recombinant protein production.The gypsy moth (Lymantria dispar) cell line, IPLB-LD-652Y, was subcloned six cell strains, IPLB-LD-652Y-a~f (LD-a ~ -f). These cell strains contain different proportions of four morphological different cells: round, spindle, squamous, and polymorphous cells. LD-a cells contain more spindle-shaped cells. The predominant cells in LD-d cells are elongate spindle cells (sensory neurve-like cells), and in LD-b, -c and -f cells are round cells. The growth rates of cell strains were faster than parental cells. These cell strains showed the same esterase isozyme pattern with that of parental cell line. LD-b and -f cells contained more lipid droplets in cytoplasm than other cell strains, while the lipid droplets were dispersed evenly in LD-b cells and aggregated in LD-f cells. All the cell strains and the parental cells were highly susceptible to LyxyMNPV-5 and LyxyNPV-2, which were isolated from the moribund larvae of L. xylina with nucleopolyhedrosis, the average infection rate was almost 98% susceptible to the viruses at 14 dpi. LD-a had high yields of polyhedra, but lower virus titer of ECV among cell strains. And the cell strains also showed a high susceptibility to fluorescent recombinant viruses, LyxyExp-EGFP and LyxyExp-DsRed. LD-a cells produced the highest amount of DsRed and EGFP, respectively, but LD-d produced the lowest recombinant proteins. The results of this study showed that sub-cloned cell strains had different morphology, characteristics and virus susceptibility compared with parental cells. We conclude that LD-a cells will be potentially useful for producing recombinant proteins. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-14T16:58:45Z (GMT). No. of bitstreams: 1 ntu-97-R95632011-1.pdf: 2430550 bytes, checksum: 9078be397fd7d156760d7f119c7c6c02 (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | 口試委員審定書............................................................................................................ i
致謝............................................................................................................................…ii 中文摘要.................................................................................................................…i ii 英文摘要..................................................................................................................... iv 壹、緒言..........................................................................................................................1 貳、往昔研究 …………………………………………………………………………3 一、 昆蟲細胞培養的發展史…………………………………..…………..3 二、 昆蟲細胞株的建立……………………………………………………4 三、 細胞株特性與鑑別……………………………………………………5 四、 吉普賽舞蛾與吉普賽舞蛾細胞株……………………………………5 五、 桿狀病毒………………………………………………………………6 (一)、桿狀病毒的特性………………………………………………...7 (二)、桿狀病毒的形式………………………………………………...7 (三)、病毒感染寄主後之基因表現時序……………………………...8 参、材料與方法............................................................................................................10 一、 供試細胞株………………………………………………………. 10 二、 母株之次選殖……………………………………………………..10 三、 細胞特性分析……………………………………………………. 10 (一) 細胞型態觀察…………………………………………………..10 (二) 細胞生長曲線…………………………………………………..11 (三) 同功異構酶比較…………………………………..………...….11 a. 細胞樣品備製………………………………………………11 b. 同功異構酶電泳……………………………………………11 c. 染色與固定………………………………………………....12 (四) 蘇丹黑染色……………………………….………………….....12 四、 供試病毒株………………………….………………………….…13 五、 病毒感受性………………………….…………………………….13 (一) 封埋體病毒數量計算……………………………………….….13 (二) 胞外病毒效價測試……………………………………………..14 (三) 螢光蛋白相對量測定…………………………………………..15 1. 細胞裂解………………………………………………..…15 2. 蛋白質濃度測定………………………………………..…15 3. 螢光蛋白之吸光值測定…………………………………..15 肆、結果................................................................................................................16 一、 吉普賽舞蛾子細胞株的建立…….......………………………….. 16 二、 吉普賽舞蛾細胞形態比較..…………………..…………………. 16 三、 吉普賽舞蛾細胞生長速率比較………………………………..…17 四、 吉普賽舞蛾細胞功異構酶分析……………………..……………18 五、 吉普賽舞蛾細胞株子株之蘇丹黑染色……………………..….. .18 六、 吉普賽舞蛾細胞細胞對野生型黑角舞蛾核多角體病毒之感受性……………………………………………………………..……18 (一) 吉普賽舞蛾細胞對 LyxyMNPV 之病毒感受性…………..18 (二) 吉普賽舞蛾細胞對 LyxyNPV-2之病毒感受性……………19 七、 吉普賽舞蛾細胞感染重組病毒之重組蛋白產量比較…………..21 (一) 吉普賽舞蛾細胞感染LyxyExp-EGFP之綠螢光蛋白產量比較 ……………………………………………………………..21 (二) 吉普賽舞蛾細胞感染LyxyExp-DsRed之紅螢光蛋白產量比較………………………………………………………………21 伍、討論........................................................................................................................22 陸、參考文獻................................................................................................................27 柒、圖表......................................................................................................................32 捌、附錄......................................................................................................................57 附錄一、同功異構酶電泳與染色……………………………………………57 | |
| dc.language.iso | zh-TW | |
| dc.subject | LyxyExp-DsRed | zh_TW |
| dc.subject | 吉普賽舞蛾 | zh_TW |
| dc.subject | IPLB-LD-652Y | zh_TW |
| dc.subject | 子細胞株 | zh_TW |
| dc.subject | 黑角舞蛾核多角體病毒 | zh_TW |
| dc.subject | LyxyExp-EGFP | zh_TW |
| dc.subject | LyxyMNPV | en |
| dc.subject | LyxyExp-DsRed | en |
| dc.subject | LyxyExp-EGFP | en |
| dc.subject | Lymantria dispar | en |
| dc.subject | cell strain | en |
| dc.subject | IPLB-LD-652Y | en |
| dc.title | 吉普賽舞蛾之次選殖細胞株IPLB-LD-652Y-a ~ -f的特性及對黑角舞蛾核多角體病毒之感受性分析 | zh_TW |
| dc.title | The characteristics of Lymantria dispar sub-cloned cell strains IPLB-LD-652Y-a~f and the susceptibility to LyxyMNPV | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 高穗生,侯豐男,蔡孟峰,張雲祥,田乃月 | |
| dc.subject.keyword | 吉普賽舞蛾,IPLB-LD-652Y,子細胞株,黑角舞蛾核多角體病毒,LyxyExp-EGFP,LyxyExp-DsRed, | zh_TW |
| dc.subject.keyword | Lymantria dispar,IPLB-LD-652Y,cell strain,LyxyMNPV,LyxyExp-EGFP,LyxyExp-DsRed, | en |
| dc.relation.page | 30 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2008-07-30 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 昆蟲學研究所 | zh_TW |
| 顯示於系所單位： | 昆蟲學系 | |
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