Construction and preliminary functional study of heterologous expression system of TRPV1 channel

VernacularTitle:TRPV1通道异源表达体系的建立和功能的初步研究
Author: Wenlan LI ; Defeng WANG ; Haiyu ZHOU ; Hongdan ZHAN ; Yifei DAI ; Weiwei ZHOU ; Li DAI ; Feng SUI ; Hairu HUO
Publication Type:Journal Article
Keywords: transient transfection; TRPV1 channel; HEK 293T cells; heterologous expression; dorsal root ganglion; gene trans-fection
From: Chinese Pharmacological Bulletin 2016;32(3):439-441
CountryChina
Language:Chinese
Abstract: Aim The TRPV1 plasmid was transiently transfected into human embryonic kidney HEK 293T cells to establish the heterologous expression system of TRPV1-channel. Methods The transfection efficiency was confirmed under fluorescence mi-croscope and the TRPV1 protein expression was identified by u-sing Western blot, and the functional characteristics of the chan-nel were studied by using the method of confocal microscopy. Results The transfection rate could reach 40% ~50%; the transfected cells were found to have a clear band at the corre-sponding position that TRPV1 should be, which indicated that TRPV1 channel protein was expressed in the transfected cells. The confocal microscopy imaging result showed that the trans-fected HEK 293T cells were activated by TRPV1 channel ago-nist. Conclusion Transient transfection of HEK 293T cells with TRPV1 channel is successfully constructed and the heterol-ogous TRPV1 channel is verified to have normal calcium-media-ting function.