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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.author | Hsiao-Chien Su | en |
| dc.contributor.author | 蘇筱茜 | zh_TW |
| dc.date.accessioned | 2021-07-01T08:11:35Z | - |
| dc.date.available | 2021-07-01T08:11:35Z | - |
| dc.date.issued | 1999 | |
| dc.identifier.citation | 黃麗芬（1996）探討菸草幾丁質酵素基因3'端poly(A)形成信號對於蛋白質產生之影響。國立台灣大學植物科學研究所碩士論文。 Abler, M. L. and Green, P. J. (1996) Control of mRNA in higher plants. Plant Mol. Biol. 32: 63-78. Beelman, C. A. and Parker, R. (1995) Degradation of mRNA in eukaryotes. Cell 81: 179-183. Belostotsky, D. A. and Meagher, R. B. (1993) Differential organ-specific expression of three poly(A)-binding protein genes from Arabidopsis thaliana. Proc. Natl. Acad. Sci. USA 90: 6686-6690. Birnboim, H. C. and Doly, J. (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl. Acids Res. 7:1513. Birnstiel, M. L., Busslinger, M. and Strub, K. (1985) Transcription termination and 3' processing: The end is in site! Cell 41: 349-359. Chang, A. C. Y., Cochet, M. and Cohen, S. N. (1980) Structural organization of human genomic DNA encoding the pro-opiomelanocortin peptide. Proc. Natl. Acad. Sci. USA 77: 4890-4894. Chuang, S. E., Chen A. L. and Chao, C. C. (1995) Growth of E. coli at low temperature dramatically increases the transformation frequency by electroporation. Nucl. Acids Res. 23: 1641. De Rocher, E. J., Vargo-Gogola, T. C., Diehn, S. H. and Green, P. J. (1998) Direct evidence for rapid degradation of Bacillus thuringiensis toxin mRNA as a cause of poor expression in plants. Plant Physiol. 117: 1445-1461. Diehn, S. H., Chiu, W. L., De Rocher, E. J. and Green, P. J. (1998) Premature polyadenylation at multiple sites within a Bacillus thuringiensis toxin gene-coding region. Plant Physiol. 117: 1433-1443. Guerineau, F., Brooks, L. and Mullineaux, P. (1991) Effect of deletions in cauliflower mosaic virus polyadenylation sequence on the choice of the polyadenylation sites in tobacco protoplasts. Mol. Gen. Genet. 226: 141-144. Guo, Z. and Sherman, F. (1995) 3'-end-formation signals of yeast mRNA. Mol. Cell. Biol. 15: 5983-5990. Hooykaas, P. J. J. (1988) Agrobacterium molecular genetics. Plant Molecular Biology Manual A4: 1-13. Ingelbrecht, I. L. W., Herman, L. M. F., Dekeyser, R. A., Van Montagu, M. and Depicker, A. (1989) Different 3' end regions strongly influence the level of gene expression in plant cells. Plant Cell 1: 671-680. Ish-Horowicz, D. and Burke, J. F. (1981) Rapid and efficient cosmid cloning. Nucl. Acids Res. 9: 2989. Jackson, R. J. and Standart, N. (1990) Do the poly(A) tail and 3' untranslated region control mRNA translation Cell 62: 15-24. Joshi, C. P. (1987) Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucl. Acids Res. 15: 9627-9640. Keller, W. (1995) No end yet to messenger RNA 3' processing! Cell 81: 829-832. Li, Q. and Hunt, A. G. (1995) A near-upstream element in a plant polyadenylation signal consists of more than six nucleotides. Plant Mol. Biol. 28: 927-934. Lowell, J. E., Fischer, R. L. and Sachs, A. B. (1992) 3'-UTR-dependent deadenylation by the yeast poly(A) nuclease. Genes Devel. 6: 2088-2099. McLauchlan, J., Gaffney, D., Whitton, J. L. and Clements J. B. (1985) The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3' termini. Nucl. Acids Res. 13: 1347-1363. MacDonald, C. C., Wilusz, J. and Shenk, T. (1994) The 64-kilodalton subunit of the CstF polyadenylation factor binds to pre-mRNAs downstream of the cleavage site and influences cleavage site location. Mol. Cell. Biol. 14: 6647-6654. MacDonald, M. H., Mogen, B. D. and Hunt, A. G. (1991) Characterization of the polyadenylation signal from the T-DNA-encoded octopine synthase gene. Nucl. Acids Res. 19: 5575-5581. Mogen, B. D., MacDonald, M. H., Graybosch, R. and Hunt, A. G. (1990) Upstream sequences other than AAUAAA are required for efficient messenger RNA 3'-end formation in plants. Plant Cell 2:1261-1272. Mogen, B. D., MacDonald, M. H., Leggewie, G. and Hunt, A. G. (1992) Several distinct types of sequence elements are required for efficient mRNA 3' end formation in a pea rbcS gene. Mol. Cell Biol. 12: 5406-5414. Newman, T. C., Ohme-Takagi, M., Taylor, C. B., and Green, P. J. (1993) DST sequences, highly conserved among plant SAUR genes, target reporter transcripts for rapid decay in tobacco. Plant Cell 5: 701-714. Ohme-Takagi, M., Taylor, C. B., Newman, T. C., and Green P. J. (1993) The effect of sequences with high AU content on mRNA stability in tobacco. Proc. Natl Acad. Sci. USA 90: 11811-11815. Ohtsubo, N. and Iwabuchi, M. (1994) The conserved 3' - flanking sequence, AATGGAAATG, of the wheat histone H3 gene is necessary for the accurate 3' -end formation of mRNA. Nucl. Acids Res. 22: 1052-1058. Ross, J. (1995) mRNA stability in mammalian cells. Microbiol. Rev. 59: 423-450. Ross, J. (1996) Control of messenger RNA stability in higher eukaryotes. TIG 12: 171-175. Rothnie, H. M., Reid J. and Hohn, T. (1994) The contribution of AAUAAA and the upstream element UUUGUA to the efficiency of mRNA 3'-end formation in plants. EMBO J. 13: 2200-2210. Rothnie, H. M. (1996) Plant mRNA 3'-end formation. Plant Mol. Biol. 32: 43-61. Russo, P., Li, W. Z., Guo, Z. and Sherman F. (1993) Signals that produce 3' termini in CYC 1 mRNA of the yeast Saccharomyces cerevisiae. Mol. Cell. Biol. 13: 7836-7849. Sanfacon, H. (1994) Analysis of figwort mosaic virus (plant pararetrovirus) polyadenylation signal. Virology 198: 39-49. Sheets, M. D., Ogg, S. C. and Wickens, M. P. (1990) Point mutations in AAUAAA and the poly(A) addition site: effects on the accuracy and efficiency of cleavage and polyadenylation in vitro. Nucl. Acids Res. 18: 5799-5805. Shinshi, H., Neuhas, J. M., Ryals, J. and Meins, F. J. (1990) Structure of tobacco endochitinase gene-evidence that different chitinase gene can arise by transposition of sequences encoding cysteine-rich domain. Plant Mol. Biol. 14: 357-368. Sullivan, M. L. and Green, P. J. (1993) Post-transcriptional regulation of nuclear-encoded genes in higher plants: the roles of mRNA stability and translation. Plant Mol. Biol. 23: 1091-1104. Sullivan, M. L. and Green, P. J. (1996) Mutational analysis of the DST element in tobacco cells and transgenic plants: Identification of residues critical for mRNA instability. RNA 2: 308-315. Wahle, E. and Keller, W. (1996) The biochemistry of polyadenylation. TIBS 21: 247-250. Wickens, M. (1990) How the message got its tail addition of poly(A) in the nucleus. Trends Biochem. Sci. 15, 389-446. Wu, L., Ueda, U. and Messing, J. (1993) 3'-end processing of the maize 27 kDa zein mRNA. Plant J. 4: 535-544. Wu, L., Ueda, T. and Messing, J. (1995) The formation of mRNA 3'-end in plants. Plant J. 8: 323-329. Xu, N., Chen, C. Y. A., and Shyu, A. B. (1997) Modulation of the fate of cytoplasmic mRNA by AU-rich elements: key sequence features controlling mRNA deadenylation and decay. Mol. Cell. Biol. 17: 4611-4621. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75049 | - |
| dc.description.abstract | 在真核細胞中，各種生理代謝作用是由基因先經轉錄作用產生pre-mRNA，pre-mRNA經修飾成為成熟的mRNA後，再經轉譯作用所產生的蛋白質來進行調控。而基因要形成成熟的mRNA，大多需要位元在基因3'非轉譯區域的poly(A) signal來指引polyadenylation的進行。由於本實驗室先前已發現poly(A) signal對蛋白質的表現量確有影響，因此對於植物中的poly(A) signal對mRNA的polyadenylation及穩定度的影響這兩方面做進一步的研究。 先將含有兩個poly(A) signals的菸草幾丁質酵素基因，構築在載體上，並經突變後，再接在載體pBI121上，以農桿菌感染菸草葉圓片的方式，產生五種不同的菸草轉殖植株，包括含有菸草幾丁質基因的兩個完整poly(A) signals (p100)、第一個poly(A) signal被破壞（pAxT）、第二個poly(A) signal被破壞（pATx）、兩個poly(A) signals均加以破壞（pAxTx）、增為四個poly(A) signals (paATAT)的不同轉殖植株。 在poly(A) signal對mRNA進行polyadenylation的影響上，發現在二個或二個以上的poly(A) signals之使用上，mRNA進行polyadenylation時似乎較偏好使用第一個poly(A) signal；但poly(A) signal的序列會進一步影響poly(A) site位置使用的頻率，研究結果發現若在二個或以上的poly(A) signals之使用上，序列完全符合AAUAAA的poly(A) signal會使得其後的poly(A) site的使用頻率大為提高。 在mRNA穩定度的研究結果發現，五種轉殖植株中，p100、pAxT轉殖株的GUS基因mRNA半衰期最長，可達三個小時左右；其他pATx、pAxTx、paATAT三種轉殖株的GUS基因的mRNA半衰期則明顯偏低，約一個多小時左右；這結果與本實驗室先前所做蛋白質短暫表現的情況相符。因此結合poly(A) sites的分佈實驗結果，我們推測mRNA越長，也就是poly(A) sites在越遠的地方形成，mRNA越穩定；而當mRNA越穩定，蛋白質的表現量就越高。 | zh_TW |
| dc.description.abstract | In eukaryotic cells, the mature mRNAs are produced by the processing of pre-mRNAs. To become mature mRNAs, the poly(A) signals in the 3' untranslated region are needed to direct the polyadenylation. In our laboratory, we have found that the poly(A) signals have influences on the gene expression. The purpose of this study is to analyze the effects of the poly(A) signals on the mRNA stability and polyadenylation in plants. Two poly(A) signals within tobacco endochitinase gene are mutated and cloned into pBI121 vectors. Five different transgenic tobaccos, including p100, pAxT, pATx, pAxTx, and paATAT, are generated by using Agrobacterium to infect leaf disks. Our study indicates that most poly(A) sites are formed at the downstream of the first poly(A) signal. Also, the poly(A) sites are affected by the sequence of the poly(A) signals if there are two or more than two poly(A) signals in the 3' untranslated region. The efficiency of the polyadenylation is improved when the sequence of the poly(A) signal is AAUAAA. The half-life of the GUS mRNA in the p100 and pAxT transgenic plants is almost three hours, which is much longer than the other tested transgenic plants whose half-life is almost one hour. Based on the position of the poly(A) sites, we conclude that the mRNA stability is controlled by the length of the mRNA, which is determined by the position of the poly(A) sites. Furthermore, the mRNA stability may be a major factor to affect the amount of GUS expression from the study of transient expression of GUS gene. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-01T08:11:35Z (GMT). No. of bitstreams: 0 Previous issue date: 1999 | en |
| dc.description.tableofcontents | 中文摘要……………………………………………………1 英文摘要……………………………………………………3 第一章 前言……………………………………………………5 第二章 材料與方法……………………………………………………11 第一節 藥品與材料……………………………………………………11 1-1 來源……………………………………………………11 第二節 載體的構築……………………………………………………12 2-1 微量DNA質體的製備……………………………………………………12 2-2 大量DNA質體的製備……………………………………………………13 2-3 限制酵素的切割……………………………………………………15 2-4 DNA凝膠電泳法……………………………………………………15 2-5 瓊脂凝膠中DNA片段的回收……………………………………………………15 2-6 DNA的粘接反應……………………………………………………16 2-7 大腸桿菌勝任細胞的製備……………………………………………………17 2-8 大腸桿菌勝任細胞的轉型作用……………………………………………………17 2-9 各突變株質體的構築……………………………………………………17 第三節 農桿菌的轉型與鑑定……………………………………………………20 3-1 農桿菌勝任細胞的製備與轉型……………………………………………………20 3-2 轉型後農桿菌的篩選與鑑定……………………………………………………21 3-3 農桿菌質體的抽取……………………………………………………22 第四節 菸草的基因轉殖與鑑定……………………………………………………22 4-1 葉圓片轉型法……………………………………………………22 4-2 菸草total RNA的抽取……………………………………………………23 4-3 RNA凝膠電泳法……………………………………………………24 4-4 GUS活性分析……………………………………………………25 4-5 蛋白質含量測定……………………………………………………25 第五節 mRNA 3'端斷點分析……………………………………………………26 5-1 RT-PCR之分析……………………………………………………26 5-2 對照組的設計……………………………………………………27 5-3 定序……………………………………………………28 5-4 定序之電泳分析……………………………………………………28 第六節 GUS基因mRNA半衰期分析……………………………………………………29 6-1 原生質體的製備……………………………………………………30 6-2 轉錄的抑制……………………………………………………31 6-3 原生質體RNA的抽取……………………………………………………31 6-4 北方轉印法……………………………………………………32 6-5 放射性探針的製備……………………………………………………34 6-6 半衰期的計算……………………………………………………36 第三章 結果……………………………………………………38 第四章 討論……………………………………………………48 參考文獻……………………………………………………56 圖表……………………………………………………63 附錄……………………………………………………93 | |
| dc.language.iso | zh-TW | |
| dc.title | 探討菸草幾丁質酵素基因的poly(A)形成信號對poly(A)形成位置和mRNA穩定度的影響 | zh_TW |
| dc.title | The Effects of Poly(A) Signals from Tobacco Endochitinase Gene in the Position of Poly(A) Sites and the Stability of mRNA | en |
| dc.date.schoolyear | 87-2 | |
| dc.description.degree | 碩士 | |
| dc.relation.page | 62 | |
| dc.rights.note | 未授權 | |
| dc.contributor.author-dept | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 植物科學研究所 | zh_TW |
| 顯示於系所單位： | 植物科學研究所 | |
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