【摘要】：[目的]為了研究CRBN基因與常染色體隱性遺傳非綜合征輕型智力發(fā)育遲滯(ARNSMR)的關系。[方法]從293T細胞中克隆了CRBN和CRBN突變體基因(CRBNm),并克隆至pc DNA-GFP、pc DNA-Flag質粒中,將重組質粒用脂質體轉染到293T細胞中,Co-IP與GFP-CRBN、CRBNm融合蛋白相互作用的293T細胞蛋白,通過PAGE膠電泳、銀染分析差異蛋白。同時我們還做了CRBN和CRBNm蛋白穩(wěn)定性和泛素化修飾研究。[結果]發(fā)現CRBN基因無義突變后沒有改變蛋白的細胞定位,但影響了相互作用蛋白,同時突變的CRBN蛋白自我泛素化修飾增強,與野生型相比更容易降解。[結論]CRBN基因的無義突變后蛋白泛素化修飾增強,細胞內相互作用蛋白發(fā)生改變,從而影響了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.
【作者單位】： 南通大學生命科學學院;江蘇師范大學生命科學學院;
【分類號】：R749.93
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本文編號：2414428