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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.author | 趙月秀 | zh_TW |
dc.date.accessioned | 2021-07-01T08:14:04Z | - |
dc.date.available | 2021-07-01T08:14:04Z | - |
dc.date.issued | 1985 | |
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Constructing maps of sites cleaved by restriction endonucleases. In Molecular Cloning. A laboratory manual (ed. T. Maniatis, E.F. Fritsch and J. Sambrook), pp. 374-381. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York. 26. Maniatis, T., E.F. Fritsch, and J. Sambrook. 1982. Southern transfer. In Molecular Cloning. A laboratory manual (ed. T. Maniatis, E.F. Fritsch and J. Sambrook), pp.382-389. Cold Spring Harbor, New York. 27. Marmur, J. 1961. A procedure for the isolation of deoxyribonucleic acid from microorganisms. J. Mol. Bid. 3:208-218. 28.Marvin, D.A. and B. Hohn. 1969. Filamentous bacterial viruses. Bacteriol. Rev. 33:172-209. 29. Mascarenhas, D., R. Kelley, and A. Campbell. 1981. DNA sequence of the att region of coliphage 434. Gene 15:151-156. 30. Miller, J. 1972. Preparation of λh80dlac phage and transformation with h80dlac. In Experiments in molecular genetics (ed. 3. Miller), pp.319-325. Cold Spring Laboratory, Cold Spring Harbor, New York. 31. Mizuuchi, M. and K. Mizuuchi. 1980. Integration recombination of bacteriophage λ: Extent of the DNA sequence involved in attachment site function. Proc. Natl. Acad. Sci. USA 77:3220-3224. 32. Mizuuchi, K., R. Weisberg, L. Enquist, M. Mizuuchi, M. Buraczynska, C. Foeller, P.L. Hsu, W. Ross, and A. Landy. 1981. Structure and function of the phageλatt site: Size, Int-binding sites and location of the crossover point. Cold Spring Harbor Symp. Quant. Biol. 45:429-437. 33. Mizuuchi, M. and K. Mizuuchi. 1985. The extent of DNA sequence required for a functional bacterial attachment site of phage lambda. Nucleic Acids Res. 13:1193-1208. 34. Nash, H.A. 1981. Integration and excision of bacteriophage λ: The mechanism of conservative site specific recombination. Ann. Rev. Genet. 15:143-167. 35. Pierson, L.S., III and M.L. Kahn. 1984. Cloning of the integration and attachment regions of bacteriophage P4. Mol. Gen. Genet. 195:44-51. 36. Rappuoli, R. and G. Ratti. 1984. Physical map of the chromosomal region of Corynebacterium diphtheriae containing corynephage attachment sites attB1 and attB2. J. Bacteriol. 158:325-330. 37. Ray, D.S. 1977. Replication of filamentous bacteriophages. In comprehensive virology (ed. H. Fraenkel-contrat and R.R. Wagner), vol.7, pp.105-178. Plenum Press, New York. 38. Rigby, P.W.J., M.D.C. Rhodes, and P. Berg. 1977. Labelling DNA to high specific activity in vitro by nick translation with DNA polymerase I. J. Mol. Biol. 113:237-251. 39. Six, E.W. and C.A.C. Klug. 1973. Bacteriophage P4: A satellite virus depending on a helper such as prophage P2. Virology 51:327-344. 40. Southern, E. 1979. Gel electrophoresis of restriction fragments. In Methods in Enzymology (ed. R. Wu), vol.68, pp.i52-i76. Academic Press, New York. 41. Toussaint, A. and A. Resibois. 1983. Phage Mu: Transposition as a life-style. In Mobile Genetic Elements (ed. J.A. Shapiro), pp.105-158. Academic Press, New York. 42. Yang, M.K. and T. T. Kuo. 1984. A physical map of the filamentous bacteriophage Cf genome. J. Gen. Virol. 65:1173-1181. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75592 | - |
dc.description.abstract | 中文摘要 為找出噬菌體Cf-11基因體上之附著位置(attachment site; att site),首先必須定出其基因體上核酸限制?之物理圖譜(physical map);將Cf-11感染柑橘潰瘍原細菌(Xanthom-onas campestris pv. citri 品系xw47)寄主,並純化其在細胞內所形成之RF DNA,以8種限制?:EcoRI、Bam HI、Hind III、Pvu II、Bgl II、Xho I、Kpn I、Sst II切割,分別定出其在RF DNA上之物理圖譜。經分析結果得知EcoRI、Bam HI、Kpn I、Sst II祇有一個剪切位置,Hind III、Pvu II、Xho I有二個位置,Bgl II有三個位置,而Xba I則無任何剪切位置,並發現此確實與另一線狀噬菌體Cf(24,42)之物理圖譜一樣,因此二者為十分相似之同類線狀噬菌體(24)。進一步將經Cf-11感染後之細菌菌株染色體以限制?剪切,藉洋菜膠電泳(agarose gel electrophoresis)分開DNA片段,並以Southern blotting方法將DNA轉移(transfer)至Gene Screen TM membrane上,然後用〔α-32P〕TTP標記(label)之RF DNA當探針(probe),作DNA-DNA雜交(hybridization)試驗及自動放射顯影(autoradiography)。將結果與用相同核酸限制?剪切之RF DNA片段對照組作比較,知:以Xho I剪切時,RF DNA可得5.85kb和1.30kb兩條感光帶,而細菌染色體則見5.85kb和另兩條分別為9.20kb與1.07kb之感光帶;以Bgl II剪切時,RF DNA可得5.13kb、1.89kb、0.47kb三條感光帶,而細菌染色體則見1.89kb、0.47kb和另兩條分別為7.70kb與6.63kb之感光帶;以Bgl II和Xho I兩種酵素一同剪切時,RF DNA可得4.22kb、1.89kb、1.10kb、0.19kb四條感光帶,而細菌染色體則見4.22kb、1.89kb、0.19kb和另兩條分別為8.34kb與1.15kb之感光帶;以Sst II和Xho I兩種酵素一同剪切時,RF DNA可得5.85kb、0.75kb、0.54kb三條感光帶,而細菌染色體則見5.85kb、0.75kb和另兩條分別為3.29kb與2.24kb之感光帶;以Kpn I和Xho I兩種酵素一同剪切時,RF DNA可得5.85kb、0.70kb、0.59kb三條感光帶,而細菌染色體則見5.85kb、0.70kb和另兩條分別為2.35kb與1.12kb之感光帶。因此得知RF DNA上之附著位置位於一約350bp的SstII-Bgl II DNA片段上,若以圖譜上唯一之EcoRI位置當作零點〔zero point),則位在距此零點約67.4%和71.8%之間的DNA片段中。最後以逢機取樣方式進一步分析,得知經噬菌體Cf-11感染後之細菌菌株染色體上之噬菌體原(prophage)插入寄主DNA之附著位置可能一定。 | zh_TW |
dc.description.abstract | Summary To localize the attachment site of bacteriophage Cf-11 genome, a physical map of the phage was constructed firstly. The RF DNAs in cells of bacterium Xanthomonas campestris pv. citri strain xw47 that had been infected with with Cf-11 were isolated, purified, and digested with restriction endonucleases EcoRI, BamHI, Hindlil, PvuII, BglII, XhoI, KpnI, and SstII. Respectively, EcoRI, BamHI, KpnI, and SstII has only one cleavage site on the RF DNA; HindIII, PvuII, and XhoI each has two sites; BglII has three sites, whereas XbaI has none. According to the results, it was found that the restriction map of Cf-11 is identical to that of another filamentous phage Cf. The chromosomal DNAs of Cf-11 infected host cells were digested with the restriction enzymes, fractioned by agarose gel electrophoresis, and transfered to Gene Screen TM membrane by Southern blotting method. Using the 332P-TTP labelled RF DNA probes to carry out DNA-DNA hybridization and autoradiography, and campared the results with the RF DNAs that had been digested with the same enzymes, it was shown: (1) with XhoI, the RF DNAs get two bands of 5.85 kb and 1.30 kb fragments, while 5.85 kb, 9.20 kb, and 1.07 kb bands in the bacterial chromosomes; (2) with BglII, the RF DNAs get three bands of 5.13 kb, 1.89 kb, and 0.47 kb fragments, while 1.89 kb, 0.47 kb, 7.70 kb, and 6.63 kb bands in the bacterial chromosomes; (3) with BglII and XhoI simultaneously, the RF DNAs get four bands of 4.22 kb, 1.89 kb, 1.10 kb, and 0.19 kb fragments, while 4.22 kb, 1.89 kb, 0.19 kb, 8.34 kb, 1.15 kb bands in the bacterial chromosomes; (4) with SstII and XhoI simultaneously, the RF DNA5 get three bands of 5.85 kb, 0.75 kb, 0.54 kb fragments, while 5.85 kb, 0.75 kb, 3.29 kb, and 2.24 kb bands in the bacterial chromosomes; (5) with KpnI and XhoI simultaneously, the RF DNAs get three bands of 5.85 kb, 0.70 kb, and 0.59 kb fragments, while 5.85 kb, 0.70 kb, 2.35 kb, and 1.12 kb bands in the bacterial chromosomes. So after the careful resolution, it was obtained that the attachment site on Cf-11 genome is located in an about 350 bp DNA fragment between the SstII and the BglII sites, that is 67.4%-71.8% from the zero point of the single EcoRI site on the map. Further analysis, when with random selection, suggested that the attachment sites on Cf-11 genome and the host chromosome may be persistent. | en |
dc.description.provenance | Made available in DSpace on 2021-07-01T08:14:04Z (GMT). No. of bitstreams: 0 Previous issue date: 1985 | en |
dc.description.tableofcontents | 目錄 中文摘要......................III 英文摘要......................V 一、緒言......................1 二、材料與方法......................5 (一)細菌與噬菌體......................5 (二)試藥......................5 (三)培養基......................6 (四)細菌和噬菌體之培養與貯存......................7 (五)緩衝液和溶液之配製......................8 (六)噬菌體RF DNA之抽取......................11 (七)細菌細胞內所有DNA之抽取......................13 (八)定Cf-11基因體上之附著位置......................15 (九)Southern blotting ......................16 (十)探針之製作......................17 (十一)DNA-DNA雜交......................21 (十二)自動放射顯影(autoradiography)......................23 (十三)Cf-11 RF DNA上各種核酸限制?之物理圖譜......................23 三、結果......................26 (一)噬菌體Cf-11基因體之物理圖譜......................26 1.核酸限制?EcoR I 、Bam HI物理圖譜之訂定......................30 2.核酸限制?Hind III物理圖譜之訂定......................30 3.核酸限制?Pvu II物理圖譜之訂定......................32 4.核酸限制?Xho I物理圖譜之訂定......................33 5.核酸限制?Bgl II物理圖譜之訂定......................34 6.核酸限制?Kpn I和Sst II物理圖譜之訂定......................38 (二)噬菌體Cf-11基因體上之附著位置......................46 1.利用各種限制?剪切找出可能之附著位置......................50 2.利用雙剪切方式詳細定出附著位置......................55 (三)進一步探討噬菌體Cf-11基因體插入寄主染色體之型式是否一定......................61 四、討論......................64 五、參考文獻......................72 | |
dc.language.iso | zh-TW | |
dc.title | 線狀噬菌體Cf-11基因體插入寄主染色體之型式 | zh_TW |
dc.title | The Mode of the Integration of the Filamentous Phage Cf-11 Genome into Host Chromosome | en |
dc.date.schoolyear | 74-2 | |
dc.description.degree | 碩士 | |
dc.relation.page | 84 | |
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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