Abstract
Transient expression in tobacco plant (Nicotiana benthamiana) is used to determine the subcellular location of a protein of interest when tagged with a reporter such as green fluorescent protein (GFP), or to mass produce proteins without making transgenic plants. The root tumor bacteria, Agrobacteria, are used as medium to introduce the target gene expression cassette into benthamiana mesophyll cells.
Keywords: Transient expression, Tobacco, Agrobacterium, Fluorescence, Leaves
Materials and Reagents
Equipment
Procedure
Recipes
References
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It depends on what you are trying to see. For observation of fluorescent proteins usually you can use small piece of a leaf and suspend it in watter. You don't have to remove an epidermis.
I have used this protocol for BiFC and it worked perfectly. I agreed with Marek. Just a piece of leaf suspended in water is good enough. Make sure you water the plants well before infiltration. I normally kept the plants in dark between 3-4 days before monitoring the fluorescence.