Probing protein targeting to plasmodesmata using fluorescence recovery after photo-bleaching

Abstract

Fluorescence recovery after photo-bleaching (FRAP) involves the irreversible bleaching of a fl uorescent protein within a specifi c area of the cell using a high-intensity laser. The recovery of fl uorescence represents the movement of new protein into this area and can therefore be used to investigate factors involved in this movement. Here we describe a FRAP method to investigate the effect of a range of pharmacological agents on the targeting of Tobacco mosaic virus movement protein to plasmodesmata.