Validation of expression and cellular localization of AtZAT12 gene deleted an EAR motif in Nicotiana benthamiana using the transient expression system

Expression and cellular localization of recombinant proteins play a key role in identifying protein functions. Plant based transient expression was developed as fast, simple, low cost, and high –level expression of genes of interest. The plant based transient expression systems reached the full expression of the gene of interest in tobacco leaves 3–4 days after infiltration. This study utilized plant transient transformation in tobacco leaves to express the stress-response marker AtZAT12 gene which deleted the EAR motif ( ZAT12ΔEAR ). The constructs of ZAT12ΔEAR inserted into the expression vectors pJNC2 containing red fluorescence marker-mCherry, thereby visualizing the localization of these EAR-deleted ZAT12 proteins in agro-infiltrated tobacco leaves. At 2 days post infiltration, the mCherry signals exhibited nuclear localization in tobacco leaf cells. So the expression and localization of ZAT12ΔEAR recombinant proteins were confirmed using transient expression before conducting a stable transformation or visualizing protein-protein interaction in plants like Bimolecular Fluorescence Complementation (BiFC).