A型核纤层蛋白前体与RanBPM相互作用的验证

摘要

Objective:Several candidates, including vascular RanBPM, from human skeletal muscle cDNA library were found in our previous study by yeast two-hybrid screening. The purpose of this study was to identify the interaction between RanBPM and prelamin A by GST pull-down analysis.Methods:Recombinant vector pET32a-RanBPM was constructed successfully and RanBPM was expressed in prokaryotic cells. Using the expressed RanBPM and prelamin A, GST pull-down assays was carried out to identify their interaction in vitro.Results: The plasmid of pET-32a-RanBPM was constructed successfully and the interaction in vitro between prelamin A and RanBPM was confirmed by GST pull-down assays.Conclusions:Our data demonstrated that RanBPM interact with prelamin A both in vitro and in vivo.%目的:本实验室前期工作通过酵母双杂交技术初步证明RanBPM是一个与A型核纤层蛋白前体相互作用的蛋白,本研究进一步通过细胞外蛋白沉降实验(GST pull-down analysis)验证其细胞外的相互作用.方法: 构建原核表达载体pET-32a-RanBPM,分别进行RanBPM和prelamin A的原核表达,通过GST pull-down实验证明两者在体外的相互作用.结果:成功构建原核表达载体pET-32a-RanBPM,通过GST pull-down实验验证了RanBPM和prelamin A在细胞外有相互作用.结论:RanBPM和prelamin A在细胞外有相互作用.