Wydro Mateusz, Kozubek Edward, Lehmann Przemysław
Institute of Plant Genetics, Polish Academy of Sciences, Poznań, Poland.
Acta Biochim Pol. 2006;53(2):289-98. Epub 2006 Apr 3.
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.
在此，我们报道了一种利用农杆菌介导的瞬时基因表达检测系统，该系统简单且可重复，通过对本氏烟草幼叶进行浸润来实现。尽管本文所描述的一些现象已有其他研究者报道过，但我们在此对其进行了进一步拓展。在用含有450 - 600微摩尔乙酰丁香酮预培养的根癌农杆菌菌株AGL0和EHA105浸润的本氏烟草最幼嫩叶片中，检测到了最高水平的瞬时绿色荧光蛋白（gfp）基因表达。尽管瞬时gfp基因表达的最高水平可能受到RNA沉默的限制，但在病毒蛋白HC - Pro存在的情况下，这种表达被完全抑制。本文所描述的瞬时表达系统可用于鉴定新的RNA沉默病毒抑制子、详细分析未鉴定基因以及在植物中进行蛋白质的工业化生产。
