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酵母双杂交技术筛选人PIF1解螺旋酶相互作用蛋白
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Journal Article
酵母双杂交技术筛选人PIF1解螺旋酶相互作用蛋白
2013
Overview
目的 应用酵母双杂交技术研究筛选与人类PIF1解螺旋酶相互作用的蛋白。方法 应用酵母双杂交技术，以人PIF1蛋白PINT功能域（1～180氨基酸）和解螺旋酶模序（167～926氨基酸）为诱饵，与HeLa细胞cDNA文库杂交，筛选能与人PIF1蛋白不同功能域相互作用的蛋白。结合生物信息学分析，一对一酵母回复性杂交及β-galactosidase实验等确定阳性克隆。结果 PIF1蛋白PINT功能域杂交共获得17个阳性克隆，生物信息学分析有3个阳性基因，它们分别是CCNDBP1、OTUD5、CAP1。而PIF1蛋白解螺旋酶模序未获得阳性克隆。结论 PIF1蛋白的PINT功能域对调控PIF1的生理功能具有非常重要的作用。
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