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dc.contributor.authorCheng-Lin Zhuoen_US
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dc.description.abstractTea is an important cash crop in the world. It is also one of the indispensable drinks in Taiwanese daily life. Previous study from our lab, we discovered unique acylated flavonol tetraglycosides in Chin Shin Oolong, which could improve gastrointestinal emptying and induce hunger. The compounds could bind to the ghrelin receptor and trigger physiological responses via the same molecular mechanism as ghrelin, and were named as teaghrelin. In addition, teaghrelin or teaghrelin-like compounds were also identified in other tea cultivars. Among them, Taiwan tea Experiment station (TTES) No. 19 and No. 20 were the breeding offsprings of Chin Shin Oolong X TTES No.12 and Chin Shin Oolong X 2202 species, respectively. However, TTES No. 19 produced teaghrelin and TTES No. 20 produced teaghrelin-like compound. Therefore, it is hypothesized that the hybridization may have caused genetic changes in the proteins involved in teaghrelin biosynthesis pathway. Thus, the objective of this study is to explore the factors involved in the teaghrelin biosynthesis pathway by using phage display system. In this study, Chin Shin Oolong seeds were collected, cultivated into seedlings, and analyzed for the biosynthesis of teaghrelin by HPLC. The results showed that, of the 100 seedlings grown, 29 seedlings did not produce teaghrelin but only produced teaghrelin-like compound. Phage display system was used to further explore factors differences between the seedlings with different ability in teaghrelin synthesis. At present, 13 peptide sequences have been selected through high-stringency panning. Dot and western blot analyses were currently used to verify the differential binding of the selected phages to different leaf extracts of tea seedlings. It is judged that four of them can specifically identify seedlings with the expression of teaghrelin. This study revealed the differences in teaghrelin production capability in the progenies of the same variety and supported that phage display systems may serve as a useful tool in analyzing the metabolic differences of tea plants.en_US
dc.description.tableofcontents第一章 前言 1 第二章 文獻回顧 3 第一節 茶葉介紹 3 一、茶種介紹 3 二、茶樹育種 4 三、茶葉功效性成分 5 第二節 茶飢素(Teaghrelin) 7 第三節 噬菌體展示技術 (phage display) 9 一、噬菌體載體的選擇 10 二、噬菌體展示技術的應用 11 第三章 材料與方法 13 第一節 實驗試劑與儀器設備 13 一、茶葉樣品 13 二、實驗藥品 13 三、實驗儀器與套組 13 第二節 實驗方法 14 一、液相層析儀樣品處理及分析 14 二、植物總蛋白萃取 14 三、噬菌體展示技術篩選(phage display) 14 四、滴度試驗(titer) 15 五、噬菌體溶菌斑個別培養 15 六、酵素結合免疫吸附分析法(ELISA) 15 七、溶菌斑提起法(plaque lifting) 16 八、聚丙烯醯胺膠體電泳(SDS Poly-acrylamide-gel-electrophoresis, SDS-PAGE) 17 九、西方墨點法(western blot) 17 十、點漬法(Dot-blotting assay) 18 第四章 結果 19 第一節 實生苗的茶飢素生合成檢測 19 第二節 噬菌體 ELISA 20 第三節 溶菌斑提起法 20 第四節 西方墨點法(western blot)與點漬法(dot blot)分析 20 第五章 討論 22 第六章 參考文獻 24 第七章 圖表 27zh_TW
dc.titleStudy on the factors involved in the teaghrelin biosynthesis pathway in Chin Shin Oolong by using the phage display systemen_US
dc.typethesis and dissertationen_US
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