請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/24224
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 黃鵬林(Pung-Ling Huang) | |
dc.contributor.author | Yi-Chun Liu | en |
dc.contributor.author | 劉怡君 | zh_TW |
dc.date.accessioned | 2021-06-08T05:18:58Z | - |
dc.date.copyright | 2005-08-01 | |
dc.date.issued | 2005 | |
dc.date.submitted | 2005-07-29 | |
dc.identifier.citation | 林雅亭. 1999. 蝴蝶蘭苯基苯乙烯酮合成酶基因轉殖之研究. 國立台灣大學園藝學研究所碩士論文.
許華欣. 1993. 蝴蝶蘭苯基苯乙烯酮合成酶cDNA之選殖及分析. 國立台灣大學園藝學研究所碩士論文. Bartley, G. E. and P. A. Scolnik. 1995. Plant carotenoids: Pigments for photoprotection, visual attraction, and human health. Plant Cell 7: 1027-1038. Batschauer, A., B. Ehmann, and E. Schafer. 1991. Cloning and characterization of a chalcone synthase gene from mustard and its light-dependent expression. Plant Mol. Biol. 16: 175-185. Busch, M., A. Seuter, and R. Hain. 2002. Functional analysis of the early steps of carotenoid biosynthesis in tobacco. Plant Physiol. 128: 439-453. Davies, K. M., S. J. Bloor, G. B. Spiller, and S. C. Deroles. 1998. Production of yellow colour in flower: Redirection of flavonoid biosynthesis in Petunia. Plant J. 13: 259-266. Elbashir, S. M., W. Lendeckel, and T. Tuschl. 2001. RNA interference is mediated by 21- and 22-nucleotide RNAs. Genes Dev. 15: 188-200. Forkmann, G. 1991. Flavonoids as flower pigment: The formation of the natural spectrum and its extension by genetic engineering. Plant Breed. 106: 1-26. Forkmann, G. and B. Dangelmayr. 1980. Genetic control of chalcone isomerase activity in flowers of Dianthus caryophyllus. Biochem. Genet. 18: 519-527. Forkmann, G. and S. Martens. 2001. Metabolic engineering and application of flavonoids. Curr. Opin. Biotech. 12: 155-160. Fukui, Y., Y. Tanaka, T. Kusumi, T. Iwashita, and K. Nomoto. 2003. A rationale for the shift in colour towards blue in transgenic carnation flowers expressing the flavonoid 3',5'-hydroxylase gene. Phytochemistry 63: 15-23. Giuliano, G., G. E. Bartley, and P. A. Scolnik. 1993. Regulation of carotenoid biosynthesis during tomato development. Plant Cell 5: 379-387. Haruta, M., M. Murata, A. Hiraide, H. Kadokura, M. Yamasaki, M. Sakuta, S. Shimizu, and S. Homma. 1998. Cloning genomic DNA encoding apple polyphenol oxidase and comparison of the gene product in Escherichia coli and in apple. Biosci. Biotechnol. Biochem. 62: 358-362. Hirschberg, J. 2001. Carotenoid biosynthesis in flowering plants. Curr. Opin. Plant Biol. 4: 210-218. Holton, T. A. and E. C. Cornish. 1995. Genetics and biochemistry of anthocyanin biosynthesis. Plant Cell 7: 1071-1083. Itoh, Y., D. Higeta, A. Suzuki, H. Yoshida, and Y. Ozeki. 2002. Excision of transposable elements from the chalcone isomerase and dihydroflavonol 4-reductase genes may contribute to the variegation of the yellow-flowered carnation (Dianthus caryophyllus). Plant Cell Physiol. 43: 578-585. Joung, J. Y., G. M. Kasthuri, J. Y. Park, W. J. Kang, H. S. Kim, B. S. Yoon, H. Joung, and J. H. Jeon. 2003. An overexpression of chalcone reductase of Pueraria montana var. lobata alters biosynthesis of anthocyanin and 5'-deoxyflavonoids in transgenic tobacco. Biochem. Biophy. Res. Comm. 303: 326-331. Joy IV, R. W., M. Sugiyama, H. Fukuda, and A. Komamine. 1995. Cloning and characterization of polyphenol oxidase cDNAs of Phytolacca ameracana. Plant Physiol. 107: 1083-1089. Kimmel, A. R. and S. L. Berger. 1987. Preparation of cDNA and the generation of cDNA libraries: Overview. Methods Enzymol. 152: 307-316. Koes, R. E., C. E. Spelt, H. J. Reif, P. J. M. van den Elen, E. Veltkamp, and J. N. M. Mol. 1986. Floral tissue of Petunia hybrida (V30) expresses only one member of the chalcone synthase multigene family. Nucleic Acids Res. 14: 5229-5239. Krol, A. R., L. A. Mur, M. Beld, J. N. M. Mol, and A. R. Stuitje. 1990. Flavonoid genes in petunia: Addition of a limited number of gene copies may lead to a suppression of gene expression. Plant Cell 2: 291-299. Lerch, K. 1988. Protein and active-site structure of tyrosinase. Prog. Clin. Biol. Res. 256: 85-98. Markham, K. R., K. S. Gould, and K. G. Ryan. 2001. Cytoplasmic accumulation of flavonoids in flower petals and its relevance to yellow flower colouration. Phytochemistry 58: 403-413. Metzlaff, M., M. O’Dell, P. D. Cluster, and R. B. Flavell. 1997. RNA-mediated RNA degradation and chalcone synthase a silencing in petunia. 88: 845-854. Moehs, C. P., L. Tian, K. W. Osteryoung, and D. DellaPenna. 2001. Analysis of carotenoid biosynthetic gene expression during marigold petal development. Plant Mol. Biol. 45: 281-293. Mol, J. N., A. R. van der Krol, A. J. van Tunen, R. van Blokland, P. de Lange, and A. R. Stuitje. 1990. Regulation of plant gene expression by antisense RNA. FEBS Lett. 1990 268: 427-30. Mori, S., H. Kobayashi, Y. Hoshi, M. Kondo, and M. Nakano. 2004. Heterologous expression of the flavonoid 3’,5’-hydroxylase gene of Vinca major alters flower color in transgenic Petunia hybrida. Plant Cell Rep. 22: 415-421. Nakayama, T. 2002. Enzymology of aurone biosynthesis. J. Biosci. Bioeng. 94: 487-491. Nakayama, T., K. Yonekura-Sakakibara, T. Sato, S. Kikuchi, Y. Fukui, M. Fukuchi-Mizutani, T. Ueda, M. Nakao, Y. Tanaka, T. Kusumi, and T. Nishino. 2000. Aureusidin synthase: A polyphenol oxidase homolog responsible for flower coloration. Science 290: 1163-1166. Nakayama, T., T. Sato, Y. Fukui, K. Yonekura-Sakakibara, H. Hayashi, Y. Tanaka, T. Kusumi, and T. Nishino. 2001. Specificity analysis and mechanism of aurone synthesis catalyzed by aureusidin synthase, a polyphenol oxidase homolog responsible for flower coloration. FEBS Lett. 499: 107-111. Napoli, C., C. Lemieux, and R. Jorgensen. 1990. Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. Plant Cell 2: 279-289. Nielsen, K. M., D. H. Lewis, and E. R. Morgan. 2003. Characterization of carotenoid pigments and their biosynthesis in two yellow flowered lines of Sandersonia aurantiaca (Hook). Euphytica 130: 25-34. Sato, T., T. Nakayama, S. Kikuchi, Y. Fukui, K. Yonekura-Sakakibara, T. Ueda, T. Nishino, Y. Tanaka, and T. Kusumi. 2001. Enzymatic formation of aurones in the extracts of yellow snapdragon flowers. Plant Sci. 160: 229-236. Schliemann, W., Y. Cai, T. Degenkolb, J. Schmidt, and H. Corke. 2001. Betalains of Celosia argentea. Phytochemistry 58: 159-165. Shahar, T., N. Hennig, T. Gutfinger, D. Hareven, and E. Lifschitz. 1992. The tomato 66.3-kD polyphenoloxidase gene: molecular identification and developmental expression. Plant Cell 4: 135-147. Shimada, Y., R. Nakano-Shimada, M. Ohbayashi, Y. Okinaka, S. Kiyokawa, and Y. Kikuchi. 1999. Expression of chimeric P450 genes encoding flavonoid-3', 5'-hydroxylase in transgenic tobacco and petunia plants. FEBS Lett. 461: 241-245. Southern, E. M. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98: 503-517. Strack, D. and W. Schliemann. 2001. Bifunctional polyphenol oxidases: Novel functions in plant pigment biosynthesis. Chem. Int. Ed. 40: 3791-3794. Su, V. and B. D. Hsu. 2003. Cloning and expression of a putative cytochrome P450 gene that influences the colour of Phalaenopsis flowers. Biotech. Lett. 25: 1933-1939. Sullivan, M. L., R. D. Hatfield, S. L. Thoma, and D. A. Samac. 2004. Cloning and characterization of red clover polyphenol oxidase cDNAs and expression of active protein in Escherichia coli and transgenic alfalfa. Plant Physiol. 136: 3234-3244. Tanaka, Y., S. Tsuda, and T. Kusumi. 1998. Metabolic engineering to modify flower color. Plant Cell Physiol. 39: 1119-1126. Van der Krol, A. R., P. E. Lenting, J. Veenstra, I. M. van der Meer, R. E. Koes, A. G. M. Gerats, J. N. M. Mol, and A. R. Stuitje. 1988. An antisense chalcone synthase gene in transgenic plants inhibits flower pigmentation. Nature 333: 866-869. Van Tunen, A. J., S. A. Hartman, L. A. Mur, and J. N. M. Mol. 1989. Regulation of chalcone flavanone isomerase (CHI) gene expression in Petunia hybrida: Coordinate, light-regulated and differential expression of flavonoid genes. EMBO J. 7: 1257-1263. Vaughn, K. C., A. R. Lax, and S. O. Duke. 1988. Polyphenol oxidase: the chloroplast oxidase with no established function. Physiol. Planta. 72: 659-665. Welle, R. and H. Grisebach. 1988. Isolation of a novel NADPH-dependent reductase which coacts with chalcone synthase in the biosynthesis of 6'-deoxychalcone. FEBS Lett. 236: 221-225. Winkel-Shirley, B. 2001. Flavonoid Biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol. 126: 485-493. Ye, X., S. Al-Babili, A. Klőti, J. Zhang, P. Lucca, P. Beyer and I. Potrykus. 2000. Engineering the provitamin A (β-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. Science 287: 303-305. Yoshida, H., Y. Itoh, Y. Ozeki, T. Iwashina, and M. Yamaguchi. 2004. Variation in chalcononaringenin 2’-O-glucoside content in the petals of carnations (Dianthus caryophyllus) bearing yellow flowers. Sci. Horti. 99: 175-186. Zhu, C., S. Yamamura, H. Koiwa, M. Nishihara and G. Sandmann. 2002. cDNA cloning and expression of carotenogenic genes during flower development in Gentiana lutea. Plant Mol. Biol. 48: 277-285. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/24224 | - |
dc.description.abstract | 為創新蝴蝶蘭的花色,本論文進行文心蘭黃色素生合成相關基因之選殖與分析,以期將來轉殖至蝴蝶蘭。以次選殖得到之金魚草aureusidin合成酶基因AmAS1 488 bp片段為探針,篩選文心蘭基因組庫,選取選殖系λOgAS9進行限制酶圖譜及南方氏雜交分析,接著進行次選殖及定序,取得文心蘭之aureusidin synthase基因,命名為OgAS1 (GenBank accession number AY858697),此基因長2,145個鹼基對,具有2個顯子及1個長507 bp的隱子,可解碼出545個胺基酸,預測之分子量為62.7 KDa。OgAS1解碼之胺基酸序列與鳳梨之多酚氧化酶 (polyphenol oxidase) 最相似,相似度達44.4%。進行南方氏及北方雜交分析得知,OgAS1非單一拷貝基因且主要在花器中進行表達。為了瞭解OgAS1之功能,將OgAS1解碼區連接於2 X CaMV 35S啟動子下游,構築得到基因表達質體,利用基因槍轉殖法進行金魚草、蝴蝶蘭及文心蘭花瓣之轉殖,並運用HPLC,分析轉殖前後花瓣內色素的組成及含量變化。未轉殖黃花金魚草花瓣萃取物之HPLC分析,在滯留時間12.57分鐘時可見一明顯波峰為aureusidin,其最大吸收波峰在405 nm。黃花金魚草花瓣單轉殖文心蘭OgAS1基因及金魚草AmAS1基因之試驗組皆可使aureusidin波峰顯著升高,白花金魚草經轉殖金魚草或文心蘭aureusidin synthase基因後,aureusidin波峰皆有下降趨勢。另白花蝴蝶蘭花瓣共轉殖蝴蝶蘭苯基苯乙烯酮合成酶 (chalcone synthase, CHS) 及金魚草AmAS1基因後,於405 nm下有一滯留時間為12.7分鐘之物質有上升的趨勢,其最大吸收波峰為370 nm,推測為chalcone之ㄧ種,待進一步分析以釐清基因功能及產物種類。轉殖文心蘭OgAS1基因則使波峰明顯下降,推測可能是因為轉入同源基因造成基因默化現象。在啟動子序列分析部份,由與資料庫比對結果得知OgAS1啟動子上游具有多個調控保守性序列,包括auxin荷爾蒙、光線、創傷、抗病機制等。 | zh_TW |
dc.description.abstract | In order to create new color of Phalaenopsis flower, a yellow pigment biosynthesis-related gene, aureusidin synthase gene, was cloned and analyzed from Oncidium. The gene fragment of aureusidin synthase from snapdragon (AmAS1) was used as probe to screen a genomic library of Oncidium. Based on restriction endonuclease maps and sequence analysis of genomic clone λOgAS9, aureusidin synthase gene from Oncidium was obtained and named OgAS1 (accession number AY858697 in GenBank). Nucleotide sequence analysis revealed that OgAS1 is 2,145 bp in length and contains two exons and one intron of 507 bp. OgAS1 encodes a polypeptide of 545 amino acid residues whose predicted molecular weight is 62.7 KDa. The deduced amino acid sequences of OgAS1 showed the highest homology with the duduced amino acid sequences of polyphenol oxidase gene from pineapple for 44.4%. Based on the result of Southern blot analysis, aureusidin synthase gene from Oncidium belongs to high-copy gene. The accumulation of mRNA was found primarily in the flowers. To understand the function of OgAS1, the open reading frame of OgAS1 was constructed into gene expression vector driven by 2 X CaMV 35S promoter. The HPLC chromatographic profile of the extract of untransformed yellow snapdragon revealed that the obvious peak at retention time 12.57 min was aureusidin which had the maximum absorption wavelength at 405 nm. Transient expression analysis of OgAS1 and AmAS1 in yellow snapdragon petals by particle bombardment displayed that the peak of aureusidin rose. However, transient expression analysis of OgAS1 and AmAS1 in white snapdragon petals showed the peak was downward. Co-transformation of chalcone synthase (CHS) gene from Phalaenopsis and AmAS1 in Phalaenopsis petals resulted that a rising peak of an unknown chemical whose retention time was 12.7 min and its maximum absorption wavelength was 370 nm. That was presumed to be a kind of chalcone. The HPLC chromatographic profile for yellow Oncidium petals transformed with OgAS1 revealed the peak of aureusidin was downward obviously. Gene silencing caused by transformed homologous gene could be one possible explanation for this phenomenon. In the promoter sequence analysis of Oncidium aureusidin synthase gene, many predicted responsive elements related to auxin, light, wounding and disease resistance were found. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T05:18:58Z (GMT). No. of bitstreams: 1 ntu-94-R92628103-1.pdf: 2567109 bytes, checksum: 57bc689b58379e80d2e33f871f9a2d59 (MD5) Previous issue date: 2005 | en |
dc.description.tableofcontents | 中文摘要……………………….…………………………………………...….1
英文摘要……………………………….…………………………………...….2壹、前言………………………...………………………………………….….4 貳、前人研究………………………………………………………………….5 一、植物黃花色生合成之代謝途徑………………………………………..5 二、花色相關基因之研究……........……………………………………......8 三、多酚氧化酶與植物色素形成之相關性............................................11 參、材料與方法……………………….………………………………………13 一、試驗材料………………………………………………………………13 (一)、植物材料…………………………………………………………13 (二)、基因庫………........………………………………………………13 二、基因之次選殖…………....…………………………………..………..13 (一)、基因組DNA之抽取……….....…………………………………13 (二)、聚合酶連鎖反應……............................……….…...……..….…14 (三)、基因片段之次選殖及定序分析………………………………...14 三、南方氏雜交分析....................................................................................16 (一) 探針之製備.....................................................................................16 (二) 南方氏雜交分析.............................................................................16 四、基因之選殖............................................................................................17 (一) 寄主細胞的製備.............................................................................17 (二) 基因庫之篩選.................................................................................17 (三) 限制酶圖譜分析與DNA定序.......................................................18 (四) 北方雜交分析.................................................................................19 (五) 基因啟動子序列分析.....................................................................20 五、轉殖質體之構築....................................................................................20 (一) OgAS1基因轉殖質體之構築.........................................................20 (二) AmAS1基因轉殖質體之構築........................................................20 (三) 基因啟動子轉殖質體之構築.........................................................21 (四) 質體 DNA 之大量製備................................................................21 六、基因槍法之暫時性表達分析................................................................22 (一) 微粒子之製備與DNA包裹...........................................................22 (二) 基因槍轉殖.....................................................................................23 七、類黃酮之萃取及HPLC分析................................................................23 (一) 類黃酮之萃取.................................................................................23 (二) HPLC分析.......................................................................................23 肆、結果............................................................................................................33 一、文心蘭aureusidin合成酶基因及cDNA之選殖與分析.....................33 (一) 基因組選殖系之篩選及限制酶圖譜分析.....................................33 (二) cDNA之篩選及分析.......................................................................34 (三) 南方氏雜交分析.............................................................................34 (四) 植株各器官之基因表現分析.........................................................35 二、文心蘭aureusidin合成酶基因啟動子序列分析.................................35 三、金魚草aureusidin合成酶基因之選殖..................................................35 四、Aureusidin合成酶基因表達分析.........................................................36 (一) 基因表達載體.................................................................................36 (二) HPLC分析.......................................................................................36 伍、討論............................................................................................................75 一、Aureusidin 合成酶基因結構特性........................................................75 二、Aureusidin 合成酶蛋白質特性............................................................75 三、Aureusidin 合成酶基因功能分析........................................................80 陸、結語............................................................................................................83 柒、引用文獻....................................................................................................84 | |
dc.language.iso | zh-TW | |
dc.title | 文心蘭黃色素生合成相關基因之選殖與分析 | zh_TW |
dc.title | Cloning and Analysis of Genes Related to Yellow Pigment Biosynthesis in Oncidium | en |
dc.type | Thesis | |
dc.date.schoolyear | 93-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 杜宜殷(Yi-Yin Do) | |
dc.contributor.oralexamcommittee | 鄭隨和(Shui-Ho Cheng),劉祖惠(Tsu-Hwie Liu),李昆達(Kung-Ta Lee) | |
dc.subject.keyword | 金魚草素合成酶,類黃色素, | zh_TW |
dc.subject.keyword | aureusidin synthase,flavonoid, | en |
dc.relation.page | 79 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2005-07-29 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 園藝學研究所 | zh_TW |
顯示於系所單位: | 園藝暨景觀學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-94-1.pdf 目前未授權公開取用 | 2.51 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。