CRBN基因突变引起轻型智力发育迟滞分子机制
发布时间：2019-01-24 11:12
【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。
[Abstract]:[objective] to study the relationship between CRBN gene and (ARNSMR) with mild intellectual retardation in autosomal recessive non-syndromes. [methods] CRBN and CRBN mutants (CRBNm), were cloned from 293T cells and cloned into pc DNA-GFP,pc DNA-Flag plasmids. The recombinant plasmids were transfected into 293T cells by liposome, Co-IP and GFP-CRBN,. The 293T cell proteins interacting with CRBNm fusion protein were analyzed by PAGE gel electrophoresis and silver staining. We also studied the stability and ubiquitin modification of CRBN and CRBNm proteins. [results] it was found that the CRBN gene did not alter the cellular localization of the protein, but affected the interacting protein, and the mutated CRBN protein was more easily degraded than the wild-type protein, and the self-ubiquification modification of the mutant protein was enhanced. [conclusion] the function of CRBN gene is affected by the enhancement of the protein ubiquification modification and the change of intracellular interaction protein after the nonsense mutation of CRBN gene.
【作者单位】： 南通大学生命科学学院;江苏师范大学生命科学学院;
【分类号】：R749.93
，
本文编号：2414428
[Abstract]:[objective] to study the relationship between CRBN gene and (ARNSMR) with mild intellectual retardation in autosomal recessive non-syndromes. [methods] CRBN and CRBN mutants (CRBNm), were cloned from 293T cells and cloned into pc DNA-GFP,pc DNA-Flag plasmids. The recombinant plasmids were transfected into 293T cells by liposome, Co-IP and GFP-CRBN,. The 293T cell proteins interacting with CRBNm fusion protein were analyzed by PAGE gel electrophoresis and silver staining. We also studied the stability and ubiquitin modification of CRBN and CRBNm proteins. [results] it was found that the CRBN gene did not alter the cellular localization of the protein, but affected the interacting protein, and the mutated CRBN protein was more easily degraded than the wild-type protein, and the self-ubiquification modification of the mutant protein was enhanced. [conclusion] the function of CRBN gene is affected by the enhancement of the protein ubiquification modification and the change of intracellular interaction protein after the nonsense mutation of CRBN gene.
【作者单位】： 南通大学生命科学学院;江苏师范大学生命科学学院;
【分类号】：R749.93
，
本文编号：2414428
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