Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/97777
標題: 利用噬菌體表面呈現系統探討青心烏龍茶飢素合成路徑之相關因子
Study on the factors involved in the teaghrelin biosynthesis pathway in Chin Shin Oolong by using the phage display system
作者: 卓誠麟
Cheng-Lin Zhuo
關鍵字: 茶;茶飢素;tea;teaghrelin
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摘要: 
茶在世界上是很重要的經濟作物,也是臺灣人日常生活中的不可或缺的飲品之一。在眾多的研究中指出茶具有許多的保健功效,在實驗室先前的研究中發現在青心烏龍品種的茶葉中含有一種獨特的醯化黃酮醇配醣體化合物,可以促進腸胃蠕動產生飢餓感。經過實驗證明可與飢餓素接受器結合,仿效飢餓素誘導的分子機制,故將其命名為茶飢素。並在其他不同茶種發現到相同或相似於青心烏龍中的醯化黃酮醇配醣體的結構,其中臺茶十九號及臺茶二十號分別為青心烏龍與臺茶12號及2022品系所育種的後代,卻分別產生茶飢素及茶飢素的相似物。因此推測不同品系的交配遺傳可能在其後代茶種的茶飢素生合成路徑相關蛋白上產生改變,因而造成茶飢素的產量或種類之不同。為了探討茶飢素生生合成的相關蛋白質,本研究利用噬菌體表面展示系統進行分析。在本實驗中,自民間採集青心烏龍間相互授粉所產生種子,並種植培養成實生苗,再以高效液相層析儀分析各株實生苗內茶飢素的生成表現。結果顯示,在培養的一百株實生苗中有二十九株實生苗不產生茶飢素,僅產生其類似物。接著利用噬菌體表面呈現系統的技術探討在這些可產生與不產生茶飢素的實生苗間,是否有茶飢素生合成路徑上的蛋白差異。目前已篩選出13條胜肽序列並進一步以點漬法與西方墨點法進行確認,判斷其中4條可特異性辨認有茶飢素表現的實生苗中的可能因子。本研究證實同一茶樹品種的實生苗後代中對於茶飢素生產能力具有明顯差異,並顯示噬菌體展示系統可作為一有效的工具用以分析此種代謝路徑差異的可能因子。

Tea 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.
URI: http://hdl.handle.net/11455/97777
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