HERG基因转染HEK293细胞及其编码通道电流的记录

目的建立HERG基因在HEK293细胞稳定表达的方法。方法利用Lipofectamine2000将pCDNA3.0-HERG转染进入HEK293细胞,通过G418筛选阳性克隆细胞系,采用免疫荧光细胞化学方法检测该蛋白的表达,用全细胞膜片钳技术测定HERG基因介导的快速激活延迟整流钾电流(Ikr)。结果免疫荧光细胞化学检测证实转染HEK293细胞中HERG通道蛋白的表达,膜片钳全细胞实验记录到Ikr。结论该方法有效地将HERG基因转染进入HEK293细胞,并稳定表达HERG通道蛋白及介导Ikr。

The transfection of HERG gene into HEK293 and the detection of HERG-encoded channel currents

Objective To establish the HERG-HEK293 cell line which can stably express the HERG channel.Methods The HEK293 cells were transfected by the pcDNA3.0-HERG using the Lipofectamine 2000,then screened by G418.The expression of HERG potassium channel proteins was detected by the immunofluorescence cytochemistry,and the rapidly activating delayed rectifier K﹢current(Ikr)was measured by the whole cell patch clamp techniques.Results The green stainings representing the HERG channel protein were detected by immunofluorescence cytochemistry,and the typical Ikr were recorded through a whole cell patch clamp techniques.Conclusions This is an effective method for transfecting the HERG gene into HEK293 and expressing the HERG channel protein and the Ikr.