Brugada综合征SCN5A基因G1712C突变的功能分析

发布时间：2018-02-01 18:03

  本文关键词： Brugada综合征 SCNA基因 GC 钠电流　出处：《南方医科大学学报》2017年02期 　论文类型：期刊论文

【摘要】：目的探讨Brugada综合征SCN5A基因新突变G1712C的电生理机制。方法采用体外定点诱变法构建携带有基因突变G1712C的p Rc/CMV-h H1的表达载体,lipo3000脂质转染法建立稳定表达p GFP-IRES-hβ1质粒的HEK293细胞系,并用G418进行筛选鉴定。分别做野生型的p Rc/CMV-h H1(h H1)和携带有基因突变G1712C的p Rc/CMV-h H1(mh H1)瞬时转染表达。进行全细胞膜片钳实验记录钠通道电流。实验结果由Patch Master以及IGOR Pro 6.0软件分析。结果 G1712C位于Na+通道蛋白α亚单位的DⅣ区S5与S6之间的P-loop上。在瞬时转染野生型的h H1的细胞系中,指令电位从-60 m V逐渐上升时,钠电流也渐变大,在-20 m V时完全激活;激活电压在-60 m V到-50 m V,反转电位在50 m V左右。在瞬时转染突变型G1712C的细胞系中,没有发现钠电流。结论野生型h H1所表达的钠通道蛋白与正常心肌细胞钠通道电生理特性相似。SCN5A基因G1712C突变导致Nav1.5通道失去功能,可能是该家系Brugada综合征的病因。
[Abstract]:Objective to investigate the electrophysiological mechanism of G1712C, a new mutation of SCN5A gene in Brugada syndrome. Methods to construct the gene mutation G1712C by site-directed mutagenesis in vitro. The expression vector of Rc/CMV-h H1. HEK293 cell lines stably expressing p GFP-IRES-h 尾 1 plasmid were established by lipo3000 lipid transfection. Using G418 to screen and identify. The wild type p Rc/CMV-h H1 1 and the gene mutation G1712 C carrying p Rc/CMV-h 1 1 were made respectively. Mh 1). Transient transfection expression. The sodium channel currents were recorded by whole-cell patch clamp experiments. The results were analyzed by Patch Master and IGOR Pro 6.0 software. G1712C is located on the P-loop between S5 and S6 in the D 鈪，

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