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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.author | 黃俊霖 | zh_TW |
| dc.date.accessioned | 2021-07-01T08:19:12Z | - |
| dc.date.available | 2021-07-01T08:19:12Z | - |
| dc.date.issued | 1996 | |
| dc.identifier.citation | 周延光1993．蔬菜主要病蟲害彩色圖鑑．41-42頁．第二版．淑馨出版社．臺北 陳立明1995．絲瓜簇葉病植物菌質體相關核醣核酸水解酵素之特性分析．國立台灣大學植物科學研究所碩士論文． Ames, G. F.-L. 1986. Bacterial periplasmic transport systems: structure, mechanism, and evolution. Annu. Rev. Biochem. 55, 397-425. Ames, G. F.-L., and Legar, H. 1992. ATP-dependent bacterial transporters and cystic fibrosis: analogy between channels and transporters. FASEBJ. 6, 2660-2666. Ames, G. F.-L., Mimura, C., and Shyamala, V. 1990. Bacterial periplasmic permeases belong to a family of transport proteins operating from E. coli to humans: traffic ATPases. FEMS Microbiol. Rev. 75, 429-446. Ankenbauer, R. G., and Nester, E. W. 1990. Sugar-Mediated induction of Agrobacterium tumefaciens virulence gene: structural specificity and activities of monosaccharides. J. Bacteriol. 172, 6442-6446. Black, L. M. 1943. Some properties of aster-yellows virus. Phytophathology 33, 2. Chen, T. A., and Jiang, Y.P. 1990. 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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76216 | - |
| dc.description.abstract | 本實驗參考Mycoplasma genitalium的ribonuclease III，設計一組PCR引子，以植物菌質體DNA為範本獲得之PCR產物，為DNA探針去篩選絲瓜簇葉病植物菌質體genomic library，結果非預期地篩選到推測含ATP-binding cassette（ABC）轉運系統（transport system）基因的選殖片段。 經序列的分析比對，此一選殖片段共3,903bp，G＋C比例為23.5mole％，符合已知植物菌植體之G＋C比例，並預測有三個完整的open reading frames（ORFs），此三個ORFs轉譯之蛋白質顯示並無如Mycoplasma及Spiroplasma利用UGA為色胺酸密碼之特性；其中ORF1及ORF2和ABC轉運系統中可結合ATP以提供主動運輸能量的ATP-binding protein有相當高的相似性；ORF3由蛋白質序列的比對並無具體之結果，但由二級結構、親水性質之分析及可能形成lipoprotein的胺基酸序列，推測ORF3可能為ABC轉運系統中對受質專一性結合的solute-binding lipoprotein；目前所知細菌的ABC轉運系統，除了獲取養份的功能外，還具有訊息傳遞、抗藥性及可能和寄生細菌之毒性有關；本文並就植物菌質體、革蘭氏陽性菌及革蘭氏陰性菌的系統，對ABC轉運系統這類蛋白質之生理功能及和寄主的交互作用進行探討。 | zh_TW |
| dc.description.abstract | Using the enriched phytoplasma DNA from diseased periwinkle total DNA as template and a pair of oligonucleotides (RN1, RN2) designed according to DNA sequence of ribonuclease III of Mycoplasma genitalium as primers, an 170-bp PCR product named RN-P was amplified. The loofah witches' broom phytoplasma λ ZAP genomic library was screened with the RN-P probe. After analyzing, and comparing the sequence of the cloned DNA fragment with the sequence in the databank, we found the DNA fragment contains the putative ATP-binding cassette (ABC) transport system genes instead of ribonuclease. That is because the cloned DNA contained the partial sequences of the primers. The cloned DNA fragment contains 3 open reading frames (ORFs). ORF1 and ORF2 have high homology with the ATP-binding proteins of the ABC transport system. There was no counterpart of ORF3 found in the protein databank. However, several properties including putative secondary structure, hydrophilicity and the amino acid sequence for acrylic site showed that ORF3 might encode a specific solute-binding lipoprotein of ABC transport system. In addition to obtain nutrients, the functions of known ABC transport system of bacteria include signal transduction, drug-resistance, and toxicity of parasitic bacteria. In this study, it possible implications to the unique traits of phytoplasmas were discussed. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-01T08:19:12Z (GMT). No. of bitstreams: 0 Previous issue date: 1996 | en |
| dc.description.tableofcontents | 中文摘要……………………………………………………i 英文摘要……………………………………………………ii 壹、緒言……………………………………………………1 貳、材料與方法……………………………………………………9 一、實驗材料來源與繁殖……………………………………………………9 二、植物及植物菌質體DNA之抽取……………………………………………………9 （一）健康及罹病植株總DNA之抽取……………………………………………………9 （二）植物菌質體DNA之純化……………………………………………………10 三、DNA探針之備製……………………………………………………11 （一）引子（primer）之設計……………………………………………………11 （二）聚合?連鎖反應（polymerase chain reaction; PCR）……………………………………………………11 （三）PCR產物之獲得……………………………………………………12 （四）PCR產物之處理……………………………………………………12 （五）載體（vector）之備製……………………………………………………13 （六）大腸桿菌DH5α勝任細胞（competent cell）之備製……………………………………………………13 （七）接合反應（ligation）……………………………………………………13 （八）轉型作用（transformation）……………………………………………………14 （九）轉型株之確認……………………………………………………14 四、植物菌質體genomic library之構築……………………………………………………14 （一）包被用細胞萃取物（in vitro packaging cell extracts）之備製……………………………………………………14 （二）DNA之酵解（digestion）……………………………………………………16 （三）接合反應（ligation）……………………………………………………16 （四）包被反應（packaging）……………………………………………………16 （五）基因庫大小之測定（titering）……………………………………………………17 五、植物菌質體genomic library之篩選……………………………………………………17 （一）基因庫的轉印（transfering）……………………………………………………17 （二）DNA探針的標識（labeling）……………………………………………………18 （三）雜合反應（hybridization）……………………………………………………18 （四）噬菌體DNA之萃取（λ ZAP DNA extraction）……………………………………………………19 六、生體內剪接作用（in vivo excision）……………………………………………………19 七、DNA序列分析（Double-stranded DNA sequence analysis）……………………………………………………20 （一）範本（template）DNA之備製……………………………………………………20 （二）引子及範本DNA之黏合（annealing）……………………………………………………21 （三）標識反應……………………………………………………21 （四）終止反應（termination reaction）……………………………………………………21 （五）DNA定序電泳膠體之備製及電泳分析……………………………………………………22 （六）DNA序列分析……………………………………………………22 八、南方轉漬雜合分析（Southern blotting）……………………………………………………23 九、植物總RNA之抽取……………………………………………………23 十、北方轉漬雜合分析（Northern blotting）……………………………………………………24 （一）RNA瓊脂凝膠電泳分析法……………………………………………………24 （二）北方轉漬法及雜合反應……………………………………………………24 參、結果……………………………………………………25 一、以日日春為宿主繁殖絲瓜簇葉病植物菌質體……………………………………………………25 二、植物菌質體DNA之純化……………………………………………………25 三、PCR DNA探針之備製……………………………………………………25 四、絲瓜簇葉病植物菌質體genomic library之構築……………………………………………………26 五、絲瓜簇葉病植物菌質體genomic library之篩選……………………………………………………26 六、重組質體選殖株特性之分析……………………………………………………27 七、植物菌質體DNA及健康日日春總DNA南方轉漬雜合分析……………………………………………………27 八、選殖株嵌入DNA片段之核?酸定序……………………………………………………27 九、3.9kb選殖片段DNA序列分析……………………………………………………28 十、植物總RNA之抽取……………………………………………………29 十一、罹病及健康日日春總RNA北方轉漬雜合分析……………………………………………………30 肆、討論……………………………………………………31 伍、參考文獻……………………………………………………35 陸、圖表……………………………………………………44 | |
| dc.language.iso | zh-TW | |
| dc.title | 絲瓜簇葉病植物菌質體可能的ABC轉運系統基因之分離及特性分析 | zh_TW |
| dc.title | Isolation and characterization of the putative ATP-binding cassette (ABC) transporter genes from the phytoplasma associated with the loofah witches' broom | en |
| dc.date.schoolyear | 84-2 | |
| dc.description.degree | 碩士 | |
| dc.relation.page | 66 | |
| 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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