期刊论文

中文

中华医学遗传学杂志

1003-9406

2016

33

4

466-470

10.3760/cma.j.issn.1003-9406.2016.04.008

国家自然科学基金

本校为其他机构

目的通过体外实验探讨SOX10基因突变E248fs致Ⅱ型Waardenburg综合征(Waardenburg syndrome,WS)发病的分子机制。 方法野生SOX10及其致病突变E248fs表达质粒瞬时转染293T细胞,用荧光素酶活性检测系统观察野生/突变SOX10蛋白对其靶基因MITF转录活性的调控作用以及突变蛋白对野生SOX10蛋白功能的影响;用生物素标记的含序列cattgtc的DNA寡核苷酸链探针分别沉淀SOX10和E248fs蛋白,检测野生/突变SOX10蛋白与靶基因MITF启动子的结合力;免疫共沉淀检测观察并分析SOX10和E248fs蛋白稳定性变化。 结果E248fs突变蛋白完全失去调控MITF启动子转录活性作用(P

OBJECTIVE: To explore the molecular mechanism of Waardenburg syndrome type II (WS2) resulting from SOX10 gene mutation E248fs through in vitro experiment. METHODS: 293T cells were transiently transfected with wild type (WT) SOX10 and mutant type (MT) E248fs plasmids. The regulatory effect of WT/MT SOX10 on the transcriptional activity of MITF gene and influence of E248fs on WT SOX10 function were determined with a luciferase activity assay. The DNA binding capacity of the WT/MT SOX10 with the promoter of the MITF gene was determined with a biotinylated double-stranded oligonucleotide probe con...