博碩士論文 91224021 詳細資訊




以作者查詢圖書館館藏 以作者查詢臺灣博碩士 以作者查詢全國書目 勘誤回報 、線上人數:17 、訪客IP:18.206.16.123
姓名 張貴雲(Kuei-Yun Chang)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 阿拉伯芥HIT1基因定序及選殖
(Identification, sequencing and cloning of Arabidopsis HIT1 gene)
相關論文
★ 阿拉伯芥突變種(hit1)之位址定位★ 阿拉伯芥之HIT1蛋白質為酵母菌Vps53p之對應物且能影響植物對高溫及水份逆境之耐受性
★ 阿拉伯芥繫鏈同源蛋白質HIT1對頂端生長之影響及熱耐受基因HIT2之遺傳定位★ 阿拉伯芥hit3遺傳位址定位與HIT1啟動子分析
★ 利用基因功能活化法研究阿拉伯芥乙烯生合成之調控機制★ 阿拉伯芥突變種hit2之位址定位
★ 利用化學遺傳法研究阿拉伯芥 revert to eto1 41 (ret41) 之功能研究★ 阿拉伯芥hit3和et突變種之生理定性及其基因定位
★ 阿拉伯芥囊泡繫鏈因子HIT1在逆境下維持內膜完整性之探討與ret8之基因定位★ 阿拉伯芥HS29之基因定位及ET參與植物耐熱機轉之探究
★ 阿拉伯芥中藉由核運輸接受器HIT2/XPO1A進行核質間運輸以促使植物耐受高溫逆境之專一分子的探索研究★ 阿拉伯芥hs49與78hs突變株之生理定性及其耐熱基因定位
★ 阿拉伯芥HIT4為不同於MOM1的新調節方式調控熱誘導染色質重組並在各個植物生長發育轉換時期表現★ 阿拉伯芥熱誘導性狀突變株R45之基因定位及HSP40參與植物耐熱機轉之探究
★ 阿拉伯芥hit4逆轉株r13及r34之基因定位與r34耐熱機轉之探究★ 蛋白質法尼脂化修飾參與植株耐熱反應
檔案 [Endnote RIS 格式]    [Bibtex 格式]    [相關文章]   [文章引用]   [完整記錄]   [館藏目錄]   [檢視]  [下載]
  1. 本電子論文使用權限為同意立即開放。
  2. 已達開放權限電子全文僅授權使用者為學術研究之目的,進行個人非營利性質之檢索、閱讀、列印。
  3. 請遵守中華民國著作權法之相關規定,切勿任意重製、散佈、改作、轉貼、播送,以免觸法。

摘要(中) 高溫對植物可以是一種致死性逆境。之前本實驗室已運用功能性遺傳學的策略,篩選出一對高溫逆境過度敏感之突變種植物,取名為hit1(heat intolerant) 。此突變植物在37℃之高溫逆境下無法存活超過4天,野生種則可。
本研究之主要目的即在於hit1遺傳位址之辨認及定序。實驗結果顯示,hit1是BAC F11F12中,阿拉伯芥登錄序號為AT1g50500之基因內,由C變成A之點突變。此推論尚須轉殖正常基因回突變株體內進行互補實驗,觀察可否恢復其抗高溫的表現型,作為最後之確認。
氨基酸序列比對的結果顯示,AT1g50500與conserve domain KOG2180有極高的相似度,而該conserve domain的代表為late golgi membrane sorting complex, subunit VPS53。Vps53p存在於酵母菌中,需與Vps52p及Vps54p形成聚合體,在細胞中主要是從trans golgi membrane將carboxypeptidase Y(CPY)運至prevacuolar / endosomal compartment(PVC)。文獻指出,CPY之結構相似於熱修克蛋白(HSP)。就此推測,HIT1在植物細胞中有可能是扮演運輸HSP的角色。除此之外,有關生理之觀察及實驗發現,HIT1基因會影響植物體內水分之平衡關係。在高濃度mannitol環境下,野生種較hit1突變植物有較優之生長表現。綜合以上結果,HIT1基因除賦予阿拉伯芥耐熱能力之外,亦涉及阿拉伯芥抵抗滲透壓的能力,其機制則有待更進一步的研究來揭露。
摘要(英) A heat-hypersensitive mutant of Arabidopsis was isolated based on its inability to survive under normally non-lethal high temperature condition. The mutant named hit1(heat-intolerant) is distinguished from wild-type plant via incubation at 37℃ for 4 days. hit1 is unable to survive under such condition for more than 4 days.
This program is to sequence genomic DNA and identify the locus on chromosome. Current results suggest that the hit1 locus is on BAC F11F12, within the gene whose login number in the Arabidopsis database is AT1g50500, and the mutation of hit1 is a single nucleotide changing from C to A.
Amino acid sequence alignment indicated that AT1G50500 contains conserve domain KOG2180, and is highly similar to the late golgi membrane sorting complex, subunit VPS53. Vps53p exists in the yeast, forming a complex with Vps52p and Vps54p, and the main function of the complex is to transport carboxypeptidase Y (CPY) from trans golgi membrane to prevacuolar / endosomal compartment (PVC). It has been indicated by some documents that CPY and HSP might have similar function in terms of structures. According to these data, it is reasonable to hypothesize that the HIT1 might play a role in transporting HSP in the plant cell. On the other hand, HIT1 could affect water balance in plants. Wild-type plants grow better than hit1 mutant at high mannitol concentrations. Taken together, HIT1 may function in sorting certain proteins whose roles are critical for plants to cope with both heat and osmotic stresses.
關鍵字(中) ★ 抗熱基因
★ 滲透壓逆境
★ 阿拉伯芥
關鍵字(英) ★ osmotic stress
★ heat stress
★ Arabidopsis
論文目次 中文摘要 -------------------------------------------------I
英文摘要 -----------------------------------------------III
誌謝 ----------------------------------------------------IV
目錄 -----------------------------------------------------V
表目錄 -------------------------------------------------VII
圖目錄 ------------------------------------------------VIII
第一章 緒論
一、 介紹 -------------------------------------------1
二、研究目的 ----------------------------------------9
第二章 材料與方法
一、 實驗材料---------------------------------------11
二、 實驗方法 --------------------------------------11
三、 實驗設計---------------------------------------17
第三章 結果
一、 T-DNA插入阿拉伯芥後表現型測試------------------18
二、核苷酸定序找尋突變點----------------------------18
三、HIT1基因的選殖 ---------------------------------19
四、阿拉伯芥HIT1基因cDNA之序列分析----------------20
五、滲透壓試驗 ----------- -------------------------21
第四章 討論
一、核苷酸定序--------------------------------------24
二、基因選殖----------------------------------------24
三、HIT1基因的功能----------------------------------25
四、生理試驗----------------------------------------28
五、異種間互補試驗----------------------------------31
第五章 結論與建議 -------------------------------------- 33
參考文獻-------------------------------------------------46
附錄 --------------------------------------------------- 53
表目錄
表1 引子組--------------------------------------------- - 43
表1 引子組(續1) ---------------------------------------- 44
表1引子組(續2)---------------------------------------- 45
圖目錄
圖1於BAC F11F12上定位HIT1基因-------------------- 34
圖2核苷酸定序圖---------------------------------------35
圖3 HIT1基因結構及突變點所在位置--------------------36
圖4 HIT1基因蛋白質序列與conserve domain比對結果----37
圖5阿拉伯芥HIT1與酵母菌Vps53p序列比對結果------- 38
圖6野生種與hit1突變種於mannitol下的發芽情形-------39
圖6野生種與hit1突變種於mannitol下的發育情形-------40
圖7野生種與hit1突變種於glucose及sucrose下的發芽、發育情形------------------------------------------- 41
圖8阿拉伯芥預測基因ATG50500與AT1G50970氨基酸序列比對結果------------------------------------------ 42
參考文獻 Alia, H.H., Chen, T., and Murata, N. (1998) Transformation with a gene for choline oxidase enhances the cold tolerance of Arabidopsis during germination and early growth. Plant Cell Environ 21: 232-239
Bell, C.J. and Ecker, J.R. (1994) Assignment of 30 microsatellite loci to the linkage map of Arabidopsis. Genomics 19: 137-144
Boston, R.S., Viitanen, P.V., and Vierling, E. (1996) Molecular chaperones and protein folding in plants. Plant Mol Biol 32: 191-222
Bohnert, H.J. and Jensen, R.G. (1996) Strategies for engineering water-stress tolerance in plants. Trends Biotech. 14: 89-97
Bajaj, S., Targolli, J., Liu, L.F., Ho, T.H.D., and Wu, R. (1999) Transgenic approaches to increase dehydration-stress tolerance in plants. Mol Breed 5: 493-503
Bouche, N. and Bouchez, D. (2001) Arabidopsis gene knockout: phenotypes wanted. Curr. Opin. Plant Biol. 4: 111-117
Burget, E.G., Verma, R., MØlhØj, M., and Reiter, W.D. (2003) The biosynthesis of L-Arabinose in plants: molecular cloning and characterization of a Golgi-Localized UDP-D-Xylose 4-Epimerase encoded by the MUR4 gene of Arabidopsis. Plant Cell 15: 523-531
Corpet, F. (1988) Multiple sequence alignment with hierarchical clustering. Nucl Acids Res. 16: 10881-10890
Cotton, R.G.H. (1993) Current methods of mutation detection. Mutat Res 285: 125-144
Carrasco, R., Almoguera, C., and Jordano, J. (1997) A plant small heat shock protein gene expressed during zygotic embryogenesis but noninducible by heat stress. J Biol Chem 272: 27470-27475
Collada, C., Gomez, L., Casado, R., and Aragoncillo, C. (1997) Purification and in vitro chaperone activity of a class I small heat-shock protein abundant in recalcitrant chestnut seeds. Plant Physiol 115: 71-77
Clough, S.J. and Bent, A.F. (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16: 735-743
Chang, C.C., Gilsdorf, J.R., Dirita, V.J., and Marrs, C.F. (2000) Identification and genetic characterization of haemophilus inffuenzae genetic island 1. Infection and Immunity 68: 2630-2637
Conibear, E., and Stevens, T.H. (2000) Vps52p, vps53p, and vps54p form a novel multisubunit complex requires for protein sorting at the yeast late golgi. Mol Biol Cell 11: 305-323
Dat, J.F., Lopez-Delgado, H., Foyer, C.H., and Scott, I.M. (1998) Parallel changes in H2O2 and catalase during thermotolerance induced by salicylic acid or heat acclimation in mustard seedlings. Plant Physiol 116: 1351-1357
Dijkwel P.P., Huijser C., Weisbeek P.J., Chua N.H., and Smeekens S.C.M. (1997) Sucrose control of phytochrome A signaling in Arabidopsis. Plant Cell 9: 583-595
Fodde, R., Losekoot, M. (1994) Mutation detection by denaturing gradient gel electrophoresis (DGGE). Hum mutat 3: 83-94
Giraudat, J., Hauge, B.M., Valon, C., Smalle, J., Parcy, F., and Goodman, H.M. (1992) Isolation of the Arabidopsis ABI3 gene by positional cloning. Plant Cell 4: 1251-1261
Gross, E., Arnold, N., Goette, J., Schwarz-Boeger, U., and Kiechle, M. (1999) A comparison of BRCA1 mutation analysis by direct sequencing, SSCP and DHPLC. Hum Genet 105: 72-78
Herman, P.L., and Marks, M.D. (1989) Trichome development in Arabidopsis thaliana. II. Isolation and complementation of the GLABROUS1 gene. Plant cell 1: 1051-1055
Hohl, M., and Peter, S. (1991) Water relations of growing maize coleoptiles. Comparison between mannitol and polyethylene glycerol 6000 as external osmotica for adjusting turgor pressure. Plant Physiol 95: 716-722
Hajdukiewicz, P., Svab, Z., and Maliga, P. (1994) The small, versatile pPZP family of Agrobacterium binary vectors for plant transformation. Plant Mol Biol 25: 989-994
Hare, D.D., Cress, W.A., Van Staden, J. (1998) Dissecting the role of osmolyte accumulation during stress. Plant Cell Environ 21: 535-553
Hong, S.W., and Vierling, E. (2000) Mutants of Arabidopsis thaliana defective in the acquisition of tolerance to high temperature stress. Proc Natl Acad Sci USA 97: 4392-4397
Hoekstra, F.A., Golovina, E.A., and Buitink, J. (2001) Mechanisms of plants desiccation tolerance. Trends Plant Sci 6: 1360-1385
Hsieh, T.H., Lee, J.T., Charng, Y.Y., and Chan, M.T. (2002) Tomato plants ectopically expressing Arabidopsis CBF1 show enhanced resistance to water deficit stress. Plant Physiol 130: 618-626
Janda, T., Szalai, G., Tari, I., and Paldi, E. (1999) Hydroponic treatment with salicylic acid decreases the effects of chilling injury in maize (Zea mays L.) plants. Planta 208: 175-180
Jander, G., Norris, S.R., Rounsley, S.D., Bush, D.F., Levin, I.M. and Last, R.L. (2002) Arabidopsis map-based cloning in the post-genome era. Plant Physiol. 129: 440-450
Keller, F., and Matile, P. (1989) Storage of sugars and mannitol in petioles of celery leaves. New Phytol. 113: 291-299
Konieczny, A., and Ausubel, F.M. (1993) A procedure for mapping Arabidopsis mutations using co-dominant ecotype-specific PCR-based markers. Plant J 4: 403-410
Krysan, P.J., Young, J.C., Tax, F.E., and Sussman, M.R. (1996) Identification of transferred DNA insertions within Arabidopsis genes involved in signal transduction and ion transport. Proc Natl Acad Sci USA 93: 8145-8150
Liu, W.D., Smith, I., Rechtzigel, K.J., Thibodean, S.N., and James, C.D. (1998) Denaturing high performance liquid chromatography (DHPLC) used in the detection of germline and somatic mutations. Nucleic Acids Res 26: 1396-1400
Markoff, A., Savov, A., Vladimirov, V., Bogdanova, N., Kremensky, I., and Ganev, V. (1997) Optimization of single-strand conformation polymorphism analysis in the presence of polyethylene glycerol. Clin Chem 43: 30-33
Markoff, A., Sormbroen, H., Bogdanova, N., Preisler-Adams, S., Ganev, V., Dworniczak, B., and Horst, J. (1998) Comparison of conformation-sensitive gel electrophoresis and single-strand conformation polymorphism analysis for detection of mutations in the BRAC1 gene using optimized conformation analysis protocols. Eur J Hum Genet 6: 145-150
McCallum, C.M., Comai, L., Gveene, E.A., Henikoff, S. (2000) Targeted screening for induced mutations. Nat Biotech 18: 455-457
Marchler-Bauer, A., Anderson, J.B., DeWeese-Scott, C., Fedorova, N.D., Geer, L.Y., He, S., Hurwitz, D.I., Jackson, J.D., Jacobs, A.R., Lanczycki, C.J., Liebert , C.A., Liu , C., Madej, T., Marchler, G.H., Mazumder, R., Nikolskaya, A.N., Panchenko, A.R., Rao, B.S., Shoemaker, B.A., Simonyan, V., Song, J.S., Thiessen, P.A., Vasudevan, S., Wang, Y., Yamashita, R.A., Yin, J.J., and Bryant, S.H. (2003) CDD: a curated entrez database of conserved domain alignments. Nucleic Acids Res. 31: 383-387
Meinle, D.W., Meinke, L.K., Showalter, T.C., Schissel, A.M., Mueller, L.A., and Tzafrir, I. (2003) A sequence-based map of Arabidopsis genes with mutant phenotypes. Plant Physiol 131: 409-418
Nam, H.G., Giraudat, J., Boer, B., Moonan, F., Loos, W.D.B., Hauge, B.M., and Goodman, H.M. (1989) Restriction fragment length polymorphism linkage map of Arabidopsis thaliana. Plant Cell 1: 699-705
Nicholas, Smirnoff. (1998) Plant resistance to environmental stress. Curr. Opin. Biotech. 9: 214-219
Ohsumi, K., Matsuda, Y., Nakajima, H., and Kitamoto, K. (2001) Cloning and characterization of the cpyA gene encoding intracellular carboxypeptidase from Aspergillus nidulans. Biosci. Biotechnol. Biochem. 65: 1175-1180
Paleg, L.G., Douglas, T.J., van Daal, A., and Keech, D.B. (1981) Proline, betaine and other organic solutes protect enzymes against heat inactivation. Plant Physiol 8: 107-114
Parsell, D.A., and Lindquist, S. (1993) The function of heat-shock proteins in stress tolerance: degradation and reactivation of damaged proteins. Annu Rev Genet 27: 437-496
Papageorgiou, G.C., and Murata, N. (1995) The unusually strong stabilizing effects of glycinebetaine on the structure and function of the oxygen-evolving photosystem II complex. Photosynth Res 44: 243-252
Parinov, S., and Sundaresan, V. (2000) Functional genomics in Arabidopsis: large-scale insertional mutagenesis complements the genome sequencing project. Curr. Opin. Biotech. 11: 157-161
Price, J., Li, T.C., Kang, S.G., Na, J.K., and Jang, J.C. (2003) Mechanisms of glucose signaling during germination of Arabidopsis. Plant Physiol 132: 1424-1438
Peters, J.L., Cnudde, F., and Gerats, T. (2003) Forward genetics and map-based cloning approaches. TRENDS in Plant Science 8: 1360-1385
Quesada V., Ponce M. R., and Micol J.L. (2000) Genetic Analysis of Salt-Tolerant Mutants in Arabidopsis thaliana. Genetics 154: 421-436
Redei, G.P. (1975) Arabidopsis as a genetic tool. Annual reviews 111-127
Ravnik-Glarak, M., Glavac, D., and Dean, M. (1994) Sensitivity of single-strand conformation polymorphism and heteroduplex method for mutation detection in the cystic fibrosis gene. Hum Mol Gen 3: 801-807
Richter, S., and Seth, A. (1998) One step direct detection of recurrent mutations in the breast cancer susceptibility gene BRAC1. Int J Oncol 12: 1263-1267
Pandey, R., and Agarwal, R.H. (1998) Water stress-induced changes in praline contents and nitrate reductase activity in rice under light and dark conditions. Physiol Mol Biol Plants 4: 53-57
Soto, A., Allona, I., Collada, C., Guevara, M.A., Casado, R., Rodriguez-Cerezo, E., Aragoncillo, C., and Gomez, L. (1999) Heterologous expression of a plant small heat-shock protein enhances Escherichia coli viability under heat and cold stress. Plant Physiol 120: 521-528
Sussman, M.R., Amasino, R.M., Young, J.C., Krysan, P.J., Austin, Phillips. S. (2000) The Arabidopsis knockout facility at the university of Wissconsin-Madison. Plant Physiol 124: 1465-1467
Tarczynski, M.C., Jensen, R.G., and Bohnert, H.J. (1992) Expression of a bacterial mtlD gene in transgenic tobacco leads to production and accumulation of mannitol. Proc Natl Acad Sci USA 89: 2600-2604
Tarczynski, M.C., Jensen, R.G., and Bohnert, H.J. (1993) Stress protection of transgenic tobacco by production of the osmolyte mannitol. Science 259: 508-510
Thomas, J.C., Sepahi, M., Arendall, B., and Bohnert, H.J. (1995) Enhancement of seed germination in high salinity by engineering mannitol expression in Arabidopsis thaliana. Plant Cell and Environ. 18: 801-806
Tax, F.E., and Vernon, D.M. (2001) T-DNA-associated duplication/translocations in Arabidopsis. Implications for mutant analysis and functional genomics. Plant Physiol 126: 1527-1538
Abebe, T., Guenzi, A.C., Martin, B., and Cushman, J.C. (2003) Tolerance of mannitol-accumulating. Transgenic wheat to water stress and salinity. Plant Physiol 131: 1748-1755
Vierling, E. (1991) The roles of heat shock proteins in plants. Annu Rev Plant Physiol Plant Mol Biol 42: 579-620
Winther, J.R., and SØrensen, P. (1991) Propeptide of carboxypeptidase Y provides a chaperone-like function as well as inhibition of the enzymatic activity. Proc Natl Acad Sci USA 88:9330-9334
Wu, S.J., Ding, L., and Zhu, J.K. (1996) SOS1, a genetic locus essential for salt tolerance and potassium acquisition. Plant Cell 8: 617-627
Wu, S.J., Locy, R.D., Shaw, J.J., Cherry, J.H., and Singh, N.K. (2000) Mutation in Arabidopsis HIT1 locus causing heat and osmotic hypersensitivity. Plant Physiol 157: 543-547
Sun, W., Bernard, C., van de Cotte, B., van Montagn, M., and Verbruggen, N. (2001) At-HSP17.6A, encoding a small heat-shock protein in Arabidopsis, can enhance osmotolerance upon overexpression. Plant J 27: 407-415
Yeh, C.H., Chang, P.L., Yeh, K.W., Lin, W.C., Chen, Y.M., and Lin, C.Y. (1997) Expression of a gene encoding a 16.9-kDa heat-shock protein, Oshsp16.9, in Escherichia coli enhances thermotolerance. Proc Natl Acad Sci USA 94: 10967-10972
Zhang, Y., Kanr, M., Price, B.D., Tetradis, S., and Makrigiorgos, G.M. (2002) An amplification and ligation-based method to scan for unknown mutations in DNA. Hum Mutat 20: 139-147
Yin, Y., Cheong, H., Friedrichsen, D., Zhao, Y., Hu, J., Mora-Garcia, S., and Chory, J. (2002) A crucial role for the putative Arabidopsis topoisomerase VI in plant growth and development. Proc Natl Acad Sci USA 99: 10191-10196
指導教授 吳少傑(Shaw-Jye Wu) 審核日期 2004-7-14
推文 facebook   plurk   twitter   funp   google   live   udn   HD   myshare   reddit   netvibes   friend   youpush   delicious   baidu   
網路書籤 Google bookmarks   del.icio.us   hemidemi   myshare   

若有論文相關問題,請聯絡國立中央大學圖書館推廣服務組 TEL:(03)422-7151轉57407,或E-mail聯絡  - 隱私權政策聲明