碩士 / 國立臺灣師範大學 / 生命科學研究所 / 98 / Trehalose, a non-reducing disaccharide compound, consists of two glucose by the α,α-1,1- glycosyl bond. It exists naturally in plants, invertebrates, fungi and most prokaryotes, like bacteria. Some reports suggest that trehalose is important for organisms to resist the stress, and it also plays an important role to make protein, membrane and DNA molecules more stable. However, there is no report suggests the protection by trehalose of DNA from UV irradiation in vivo. Our objective is to confirm the protection of DNA by trehalose from UV in vivo. We quantify the formation of cyclobutane pyrimidine dimers (CPDs) in DNA under multiple dosage of UV irradiation by alkaline gel electrophoresis with pyrimidine dimer specific T4 pdg digestion. Formation of CPDs is an important index to determine the level of DNA damage, and we analyze the relationship between DNA damage and trehalose. First, we extract genomic DNA from E .coli and mix with multiple dosage of trehalose, then make the mixture expose to UV light. We found the levels of intact DNA decreased with increasing UV dosage, and the DNA mixed with 100 mM trehalose showed more intact than that mixed with less trehalose. We make E. coli accumulate intracellular trehalose in vivo by induction of trehalose synthase, and then exposed to UV irradiation, followed by analysis. The results show that the cells with accumulated trehalose formed 22 CPDs/Mbs under UV dosage of 720 J/m2, and cells without intracellular trehalose formed 46 CPDs/Mbs. According to these results, trehalose not only protect DNA from UV in vitro, but also perform protection of DNA against structure damage from UV irradiation in vivo.
| Identifer | oai:union.ndltd.org:TW/098NTNU5112015 |
| Date | January 2010 |
| Creators | Hsueh-Hua Lu, 呂學樺 |
| Contributors | Guan-Chiun Lee, 李冠群 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 62 |
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