【摘要】 Ndi1p(internal NADH脱氢酶)是酵母线粒体电子传递链的重要组成部分,参与酵母线粒体呼吸、凋亡等多种生理活动.本文成功建立了酵母Ndi1p突变表达文库,随机测序表明,每个基因平均含有2个突变.利用本文库进行了Ndi1p温度敏感突变筛选,获得了一定数量的温度敏感型菌株,并对温度敏感机理做了简单探索.结果表明,温度敏感酵母在需要Ndi1p脱氢酶活性的培养基上对温度敏感;有趣的是,这些温度敏感株细胞如果在30℃生长但在37℃测试不表现出温度敏感性,这暗示高温影响温度敏感Ndi1p的生成,正常温度下Ndi1p正确构象一旦生成则高温不能引起Ndi1p变性.Ndi1p突变表达文库的建立对于Ndi1p参与的细胞呼吸、凋亡等过程的机理研究将有一定意义.更多还原

【Abstract】 Ndi1p(internal NADH dehydrogenase) is a major component of complex I that is involved in biological processes of respiration chain reactions and apoptosis.A PCR-based mutant expression library of yeast Ndi1p was constructed.The average mutation rate was about 2 changes per gene copy.A screen for Ndi1 temperature sensitive strains resulted in several isolated Ndi1-TS mutants(37℃).Once the mutant yeast was grown at 30℃on glycerol plates,which require Ndi1p NADH dehydrogenase activity,the respiration appeared to be normal as the wild type and no longer sensitive to the temperature shift to 37℃.This implied that Ndi1 malformation was originated at high temperatures.Our mutation library and the obtained mutants could be used for the screening studies in Ndi1-related respiration,as well as in apoptosis research.更多还原