Optimization of transient Agrobacterium-mediated gene expression system in leaves of Nicotiana benthamiana.
Optimization of transient Agrobacterium-mediated gene expression system in leaves of Nicotiana benthamiana.
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DOI：
10.18388/abp.2006_3341
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发表时间：
2006-04
影响因子：
1.7
通讯作者：
M. Wydro;E. Kozubek;P. Lehmann
中科院分区：
文献类型：
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作者：
M. Wydro;E. Kozubek;P. Lehmann
在这里，我们报告了一种简单且可重复的农杆菌介导的瞬时基因表达分析系统，该系统利用了烟草幼叶的渗透。虽然本文中描述的一些现象已经被其他研究人员报道过，但我们在这里对它们进行了进一步的发展。在450-600微米的乙酰丁香酮存在下，根癌农杆菌AGL0和EHA105菌株渗入的本底雪莲幼叶中瞬时GFP基因的表达水平最高。虽然瞬时GFP基因表达的最大水平可能受到RNA沉默的限制，但在病毒蛋白HC-Pro的存在下，它完全被抑制。本文所述的瞬时表达系统可用于识别新的RNA沉默病毒抑制子，用于对未知基因的详细分析，也可用于植物蛋白质的工业化生产。
Here we report on a simple and reproducible system of Agrobacterium-mediated transient gene expression assay that utilizes infiltration of young Nicotiana benthamiana leaves. Although some of the phenomena described in this paper have been already reported by other researchers, here we have further developed them. The highest level of transient gfp gene expression was detected in the youngest leaves of N. benthamiana infiltrated with A. tumefaciens strains AGL0 and EHA105 precultured in the presence of 450-600 microM acetosyringone. Although the maximum level of transient gfp gene expression was restricted presumably by RNA silencing, it was completely suppressed in the presence of the viral protein HC-Pro. The transient expression system described here can be used to identify new viral suppressors of RNA silencing, for detailed analysis of unidentified genes and for industrial production of proteins in plants as well.