【摘要】 为确定一种定量研究酵母体内蛋白质 -蛋白质之间相互作用的简便、快捷的方法 ,为抗癌小分子化合物药物筛选提供一条可行性途径。本文选择P5 3蛋白与SV4 0大T抗原为靶蛋白对 ,首先用酵母双杂交系统 (LiAc法质粒共转化酵母细胞 )方法定性证明了两者之间存在相互作用 ,然后通过α -半乳糖苷酶活力定量测定了相互作用力的大小 ,并确定了最佳酶活测定时间 ,并与 β-半乳糖苷酶活力测定进行了比较 ,认为α-半乳糖苷酶活力定量测定是研究蛋白质 -蛋白质间相互作用的一种更加简便快捷的方法。更多还原

【Abstract】 To identify a convenient method of studying protein-protein interaction in Saccharomyces cerevisiae and find an optimal way for anti-cancer drug screening,Protein53 and Simian virus 40(SV40) are used target proteins in the paper.The interaction between Protein 53 and Simian virus 40(SV40) was qualitatively proved using LiAc-mediated yeast transformation (yeast two-hybrid system).Then the interaction of the two proteins was quantitatively proved by means of α-Gal assay and identified the optimal time of yeast growth of α-Gal assay.At the same time,The α-Gal and β-Gal assay method were compared and we think that α-Gal quantitative assay is a more convenient method than β-Gal assay at the aspect ofstudying the interaction of protein-protein.It is potentially used as drug screening of small molecular compounds.更多还原