The Elongation Factor 1 Alpha Promoter Drives the Functional Expression of Kir2A in Plutella xylostella Cells

文献类型： 外文期刊

第一作者： Wang, Yinna

作者： Wang, Yinna;Ma, Haihao;Zhou, Xiaomao;Wang, Yinna;Zhou, Xiaomao;Ma, Haihao;Liu, Zheming;Zhao, Piao;Liu, Jia;Zhu, Hang;Zhou, Yong;Man, Yilong;Ma, Haihao;Liu, Zheming;Zhao, Piao;Liu, Jia;Zhu, Hang;Zhou, Yong;Man, Yilong;Zhou, Xiaomao;Ma, Haihao;Liu, Zheming;Zhao, Piao;Liu, Jia;Zhu, Hang;Zhou, Yong;Man, Yilong;Zhou, Xiaomao

作者机构：

关键词： Plutella xylostella; EF1 alpha promoter; Kir2 channel; cell line

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：4.9； 五年影响因子：5.7 ）

ISSN： 1661-6596

年卷期： 2025 年 26 卷 7 期

页码：

收录情况： SCI

摘要： Cell lines and their corresponding expression plasmids are extensively utilized in the study of insect physiology and pathology. In this research, four single-cell cultured lines (Px4-1 to Px4-4) of Plutella xylostella were established from eggs. The promoter for the P. xylostella elongation factor 1 alpha (PxEF1 alpha), known for its high driving activity in cells, was cloned and used to construct expression plasmids. Dual-luciferase activity assays and EGFP expression analyses demonstrated that the PxEF1 alpha promoter exhibited the strongest driving activity in Px4-2 cells, comparable to that of the immediate-early 1 promoter associated with the homologous region 5 enhancer (AcIE1hr5) from the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV). In contrast, the driving activity of PxEF1 alpha in cells derived from Spodoptera frugiperda, Trichoplusia ni, and Helicoverpa armigera was lower. Furthermore, the PxEF1 alpha promoter was successfully employed to drive inward rectifier potassium 2A (Kir2A) expression in Px4-2 cells. The electrophysiological properties of the insect Kir2A channel were successfully characterized for the first time. It was observed that the PxKir2A channel possesses typical inward rectifier potassium channel properties and can be inhibited by nanomolar concentrations of VU625 and VU590. This study offers a novel approach for the expression and investigation of foreign gene function in insect cells and provides a valuable tool for the in-depth study of key biomolecules in P. xylostella.

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