玉米转录因子ABP9瞬时表达体系的建立

Establishment of Maize Transcription Factor ABP9 Transient Expression System

[目的]研究玉米抗逆相关转录因子ABP9在玉米抗逆应答中的功能及其表达调控机理。[方法]以2叶期玉米叶片为材料分离原生质体,通过PEG介导的转化方法,建立玉米叶肉原生质体瞬时表达ABP9的系统。利用该系统,通过反转录PCR和Western blot分别检测ABP9genomic∷FLAG融合基因在原生质体中的转录和翻译;并通过转化ABP9∷GFP融合表达载体和GFP荧光检测,对ABP9进行亚细胞定位。[结果]玉米原生质体瞬时表达ABP9系统中,ABP9genomic∷FLAG融合基因能够正确转录出mRNA并翻译出蛋白质;利用该系统进行亚细胞定位结果显示ABP9定位于细胞核中。[结论]玉米原生质体瞬时表达ABP9体系的建立,为进一步探究玉米中ABP9的表达和调控提供了良好的实验系统。

[ Objective ] To establish a transient expression system for analyzing the function and regulation of ABP9 （ ABRE Binding Protein 9 ） in maize stress responses. [ Method] To this end, protoplasts isolated from leaves of two-leaf-stage maize plants were transformed with ABP9 constructs by a PEG-mediated procedure, and RT-PCR and Western blot analysis were used to detect the transcript and protein of ABP9genomic ： ： FLAG fusion in transformed maize protoplasts, and the ABP9 ： ： GFP fusion was expressed to analyze the subceUular localization of ABP9. [ Result ] The results showed that the ABP9genomic ：： FLAG fusion was correctly transcribed and translated, and the ABP9 ：： GFP fusion was expressed and localized in the nucleus as expected in the optimized maize mesophyll protoplast transient expression system. [ Conclusion] These data indicate that a maize transient expression system suitable for studying ABP9 expression and regulation has been established.