臺灣博碩士論文加值系統

HTL1是一個和RSC(remodel the structure of chromatin)複合物具有密切關係的酵母菌基因，部份RSC複合物成員和HTL1具有遺傳交互關係，利用抗原決定基標記HTL1 基因的菌株，我們發現HTL1和RSC、STH1具有共同免疫沉澱關係而且HTL1作用位置在細胞核。另外，最近的文獻指出，STH1和RSC8可控制一個代謝相關基因-CHA1的表現，在非誘發狀態下，sth1或rsc8突變株會大量表現CHA1基因，所以推測RSC複合物在非誘發狀態時會抑制CHA1的表現。由於這被認為是RSC複合物可能直接作用的基因，而且其真正作用機制並未完全被了解，所以我們嘗試探討HTL1是否也會調控CHA1基因的表現。利用htl1基因剔除株的研究發現，非誘發狀態下，htl1基因剔除株的CHA1轉錄活性明顯低於野生型菌株，而htl1基因剔除株仍然可以發生CHA1基因誘發，所以我們推測HTL1可能參與CHA1基因表現調控且突變株的效果與sth1或rsc8不同。此外我們也發現，山梨醇(sorbitol)可以部分拯救htl1基因剔除株高溫敏感，而且htl1基因剔除株具有對calcofluor white敏感的細胞壁完整性缺陷的特徵。

Yeast Htl1p was found to be functionally related to RSC complex by genetic analysis. In addition to the association of Htl1p and Rsc8p, other components may also interact with Htl1p. Using HA tagged strain; we show that Sth1p and Rsc8p can interact with Htl1p by co-immunoprecipitation. By immunofluorescent staining, Htl1p was found in the nucleus. In recent report, STH1 and RSC8 have been shown to be required for proper transcriptional regulation and nucleosome positioning in CHA1 promoter. Here we show that the htl1 disruption strain prolonged the repression state in the absence of serine. In the presence of serine, the de-repression of the CHA1 expression is similar to that of wild-type yeast. The observation that htl1 disruption strain was demonstrated to be sensitive to calcofluor white dye which binds to chitin of the cell wall and the temperature sensitive phenotype of htl1 mutant can be partially rescued by sorbitol suggests that HTL1 involved in the mutation of the cell wall integrity.

中文摘要---------------------------------------------------------------------------------------- 1
英文摘要---------------------------------------------------------------------------------------- 2
緒論---------------------------------------------------------------------------------------------- 3
實驗材料---------------------------------------------------------------------------------------- 11
實驗方法---------------------------------------------------------------------------------------- 14
實驗結果---------------------------------------------------------------------------------------- 23
討論-- ------------------------------------------------------------------------------------------- 28
參考文獻---------------------------------------------------------------------------------------- 33
結果圖表---------------------------------------------------------------------------------------- 38

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