Bimolecular Fluorescence Complementation (BIFC) Protocol for Rice Protoplast Transformation

Kun  Wang; Ying Liu; shaoqing Li

Bimolecular Fluorescence Complementation (BIFC) Protocol for Rice Protoplast Transformation

2013

Kun Wang | Ying Liu | shaoqing Li

After the plant cells are removed the cell walls by digestive enzyme, the plant protoplasts still have good cell activity. The protoplasts can be used to transiently express proteins of target genes in living plant cells through polyethylene glycol (PEG) mediated transformation. The purpose of this method is to employ the rice protoplasts and Green fluorescent protein (GFP) as an experimental system to observe the protein interactions in vivo. Meanwhile a 505~530 nm emission filter is used in confocal microscope to eliminate the interference of the autofluorescence from plant cells. The phenomenon of plant cell body spontaneous fluorescence can be eliminated by confocal observation.

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