新抑癌基因SLR417之細胞功能探討: I. 利用酵母雙雜交 
系統找尋SLR417之交互作用蛋白質

癌症是目前世界上相當流行的重大疾病，其中肺癌不論在全球或是台灣死亡率都是排行第一。至今多數的研究指出，癌症的發生與細胞中多種基因發生突變相關。在本實驗室先前與國衛院黃奇英老師的實驗中，利用基因晶片比較肺癌病患的腫瘤組織與相連的正常組織基因的表現量，發現到數個基因在二者的表現程度差異較大，其中SLR417在腫瘤組織的表現量相對於正常組織是偏低的。綜合基因晶片、Q-RT-PCR分析的結果與SLR417的相關資料，我們假設SLR417很可能是一個新的抑癌基因。為了研究這一個新穎的基因，我們利用酵母菌雙雜交法這個研究蛋白質與蛋白質交互作用的方法，試著在細胞體內尋找有意義的結合蛋白。我們在含GAL4 BD (結合區) 的質體pGBKT7上，插入了全長的SLR417，使得GAL4 BD-SLR417融合蛋白成為一個餌，並向廠商購買睪丸的cDNA基因庫做為有待誘捕的獵物。在酵母菌雙雜交實驗後，我們在TDO (缺乏His、Leu、Trp及Ade等四種營養素) 的培養盤中挑到717個雜交後的酵母菌株，其中有54個菌株在X-

關鍵字

酵母菌雙雜交

並列摘要

Cancer is one of the human diseases with high prevalence in the world, among them, lung cancer is the leading cause of cancer death around the world as well as in Taiwan. The previous evidence proved that cancer has a relationship with multi-gene mutation. In our previous studies, using lung cancer tissue and its’ adjacent non-tumor part with microarray, we found some genes with different expression level in both part. SLR417 is one of those genes which is down-regulated in tumor part as compared to adjacent normal part. Combined the microarray and Q-PCR analysis data and some SLR417 relating information, we hypothesize SLR417 maybe a novel tumor suppressor gene. To study this novel gene, we use yeast two hybrid assay which is one method to study protein-protein interaction in vivo to find those meaningful interaction protein. We constructed the full-length human SLR417 gene to fuse in-frame with the GAL4 DNA binding domain as bait, a commercial human testis cDNA library as prey. Our results show that there are 717 colonies selectively grew on QDO plates (Ade.His.Leu.Trp nutritionally deficient plates) and 54 colonies among those could also activate

並列關鍵字

yeast two hybrid

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