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| 基因突变对RUNX2蛋白亚细胞定位的影响 | |
| 引用本文： | 梁国健,谢宝仪,轩东英. 基因突变对RUNX2蛋白亚细胞定位的影响[J]. 广东牙病防治, 2011, 19(2): 68-71 |
| 作者姓名： | 梁国健 谢宝仪 轩东英 |
| 作者单位： | 阳江市人民医院口腔科;广东省口腔医院;南方医科大学附属口腔医院; |
| 基金项目： | 广东省自然科学基金博士启动课题资助(9451026003003833); 高等学校博士学科点专项科研基金(20100101120114) |
| 摘 要： | 目的拟建立包含基因突变位点的真核表达载体,通过细胞转染了解RUNX2突变体亚细胞定位的改变。方法从患者全血中提取总RNA,通过反转录酶-聚合酶链反应获得目的片段,定向克隆获得包含基因突变位点及野生型位点的pEGFP-C1-RUNX2真核表达载体,通过瞬时转染NIH3T3成纤维细胞,显微镜下观察突变蛋白的亚细胞定位变化。结果成功构建包含突变位点及野生位点的真核表达载体pEGFP-C1-RUNX2,转染细胞24 h后,在荧光显微镜下观察转染细胞内的荧光蛋白表达情况,转染野生型RUNX2基因的细胞仅在胞核内表达绿色荧光,胞浆内无绿色荧光表达;分别引入突变位点c.674 G〉A及c.673 C〉T的表达载体转染的两组细胞,在胞浆胞核内均表达绿色荧光。结论不同的突变位点可以造成RUNX2蛋白亚细胞定位的改变,蛋白在核内累计表达量不足可能是造成单倍体不足的原因之一。 |
| 关 键 词： | RUNX2基因 突变 亚细胞定位 颅骨锁骨发育不良 |
Effect of the RUNX2 Gene Mutations on Subnuclear Localization of the RUNX2 Protein Mutants |
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| LIANG Guo-jian,XIE Bao-yi,XUAN Dong-ying. Effect of the RUNX2 Gene Mutations on Subnuclear Localization of the RUNX2 Protein Mutants[J]. Journal of Dental Prevention and Treatment, 2011, 19(2): 68-71 | |
| Authors: | LIANG Guo-jian XIE Bao-yi XUAN Dong-ying |
| Affiliation: | LIANG Guo-jian1,XIE Bao-yi,XUAN Dong-ying.1Stomatological Department of People's Hospital in Yangjiang City,Yangjiang 529500,China |
| Abstract: | Objective To observe the effect of the RUNX2 gene mutations on subnuclear localization of the RUNX2 mutant protein by transfection.Methods Total RNA was extracted and reverse-transcribed from blood of a patient with cleidocranial dysplasia,and the eukaryotic expression vector pEGFP-C1-RUNX2 was constructed by subcloning the objective fragments.The subnuclear localization of RUNX2 wild and mutant protein was observed by transfected into NIH3T3 fibroblast cells.Results The vector pEGFP-C1-RUNX2 was constructe... |
| Keywords: | RUNX2 gene Mutation Subnuclear localization Cleidocranial dysplasia |
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