【摘要】 [目的]为了研究CRBN基因与常染色体隐性遗传非综合征轻型智力发育迟滞(ARNSMR)的关系。[方法]从293T细胞中克隆了CRBN和CRBN突变体基因(CRBNm),并克隆至pc DNA-GFP、pc DNA-Flag质粒中,将重组质粒用脂质体转染到293T细胞中,Co-IP与GFP-CRBN、CRBNm融合蛋白相互作用的293T细胞蛋白,通过PAGE胶电泳、银染分析差异蛋白。同时我们还做了CRBN和CRBNm蛋白稳定性和泛素化修饰研究。[结果]发现CRBN基因无义突变后没有改变蛋白的细胞定位,但影响了相互作用蛋白,同时突变的CRBN蛋白自我泛素化修饰增强,与野生型相比更容易降解。[结论]CRBN基因的无义突变后蛋白泛素化修饰增强,细胞内相互作用蛋白发生改变,从而影响了CRBN基因的功能。更多还原

【Abstract】 [Objective]To explore the relationship of CRBN gene with autosomal recessive nonsyndromic mental retardation(ARNSMR).[Methods]CRBN and mutant CRBN(CRBNm) genes were cloned from 293 T cell line.After constructed to pc DAN-GFP and pc DNA-Flag vectors,transfected vectors of CRBN and CRBNm to 293 T cell line with Lipo 2000.Co-IP proteins that can bind GFP-CRBN or CRBNm from 293 T cell lysis solution;After electrophoresed by PAGE gel,analyzed diversity bands using silver staining method.At the same time,proteins stability and ubiquitin modification were detected.[Results]After the premature stop codons of CRBN,CRBNm gene localization does not change,but combined the special interaction proteins.Mutant CRBN can increase ubiquitin modification and is more readily degraded than WT CRBN.[Conclusion]These changes may influence CRBN gene functions and contribute to ARNSMR.更多还原