Helm Jutta Maria, Dadami Elena, Kalantidis Kriton
Institute of Molecular Biology and Biotechnology, Heraklion, Greece.
Methods Mol Biol. 2011;744:97-108. doi: 10.1007/978-1-61779-123-9_7.
Agroinfiltration is a very fast and powerful method to express in planta any sequences in a transient fashion. Agroinfiltration has proven very useful for the overexpression of proteins in the infiltrated zone when a short-term effect can be informative. However, it has been a real success story in the induction of local and eventually systemic silencing. Here, we describe the use of agroinfiltration for the induction of local silencing of an endogene or a transgene, for the systemic silencing of a transgene and for co-infiltration assays. We also provide protocols for the evaluation of the efficiency of the assay, by detecting the specific siRNAs characteristic of RNA silencing and measuring the effects on the target sequences.
农杆菌浸润法是一种非常快速且有效的方法,可在植物中瞬时表达任何序列。当短期效应具有参考价值时,农杆菌浸润法已被证明对于在浸润区域过表达蛋白质非常有用。然而,它在诱导局部乃至系统沉默方面取得了巨大成功。在此,我们描述了农杆菌浸润法在诱导内源基因或转基因的局部沉默、转基因的系统沉默以及共浸润分析中的应用。我们还提供了通过检测RNA沉默特有的特异性小干扰RNA(siRNA)并测量对靶序列的影响来评估分析效率的方案。