| Summary: | 碩士 === 元培科技大學 === 醫學檢驗生物技術研究所 === 99 === Calorie restriction extending life span observed in a wide variety of organisms including vertebrates and invertebrates. We used the unicellular protist Trichomonas vaginalis as a model to study 14-3-3 regulate the calorie restriction in T. vaginalis. In this study ,we focusd on the 14-3-3 protein. 14-3-3 protein participates in many important cellular processes. We have been able to analyze the 14-3-3 cDNA of T. vaginalis and it was named Tv_14-3-3A. We obtained the rabbit anti-14-3-3 antibody by using Tv_14-3-3A recombinant protein. By using its antibody, we are able to study the role played by the 14-3-3 protein inside the T. vaginalis under low glucose concentration as the calorie restriction. In this study, we showed T. vaginalis cultured in YIS medium without add glucose can be extended life span than cultured in YIS medium add 1% glucose. We used real-time PCR for analyze the 14-3-3 gene expression of T. vaginalis. We found that 14-3-3 mRNA expression was significantly decreased under calorie restriction in 36 hours. Similar result was found on protein levels by using a rabbit anti-14-3-3 antibody. Western blot analysis was observed Tv_14-3-3 protein expression decreased under calorie restriction in 48 hours. Therefore, we conclude that 14-3-3 may be involved in caloric restriction pathway.
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