2.4 Subcellular localization
To analyze the subcellular location of proteins in vivo , full-length copies of PpVIN2and PpINH1 were cloned and inserted into pCAM35s-GFP plasmids at the KpnI-XbaI and BamHI-KpnI sites, respectively, generatingPpVIN2 -pCAM35s-GFP and PpINH1 -pCAM35s-GFP fusion vectors. The constructs PpVIN2 -pCAM35s-GFP, PpINH1 -pCAM35s-GFP, and the empty vector pCAM35s-GFP were transformed into Agrobacteriumstrain GV3101, then injected into the leaf epidermis of Nicotiana benthamiana . After incubation for two days, the inoculated leaves were examined by fluorescence microscopy using a laser scanning confocal microscope (FV10-ASW, OLYMPUS, Japan).