HERG基因转染HEK293细胞及其编码通道电流的记录

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

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。

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.