A型流感病毒H5N1的M2离子通道稳定细胞株的建立

Establish Stable Cell Line to Express M2 Ion Channel of Influenza a Virus H5N1

摘要:

A型流感病毒H5N1的M2离子通道(H5M2)基因经优化后由人工合成, 适合于哺乳动物细胞中表达。通过酶切克隆于pcDNA4质粒, 并在HEK293细胞中建立稳定细胞株。Western blotting和免疫荧光证实H5M2在稳定细胞中只有在四环素诱导下才能表达, 并经膜片钳证实在HEK293细胞中表达的H5M2具有H+通道活性, 为M2离子通道功能的研究和M2离子通道阻断剂筛选方法的建立提供了参考。

Abstract:

The M2 ion channel protein is an important target against influenza A virus. In this study, H5N1 influenza A virus M2 ion channel (H5M2) gene was cloned into pcDNA4 vector. The HEK293 stable cell line expressing H5M2 was successfully established. The expression of H5M2 ion channel protein was induced only by tetracycline and confirmed by imuunofluorescence and Western blot. The ion channel activity of H5M2 was confirmed by whole cell patch-clamp recording. Fifty mmol per liter amantadine blocked the H5M2 channel conductance completely in HEK293 cells. This stable cell line may provide a model for screening inhibitors of M2 ion channel.