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Study on the factors involved in the teaghrelin biosynthesis pathway in Chin Shin Oolong by using the phage display system
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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.
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