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. 2013 Nov 19;8(11):e79622. doi: 10.1371/journal.pone.0079622

Figure 1. A schematic presentation of the bidirectional promoter (P85–P87), promoter-reporter constructs and transient expression analysis using two reporter genes (GFP and GUS) in both orientations.

Figure 1

(A) A schematic map of the bidirectional promoter (P85–P87) located in the intergenic region (IR, 1285 bp, genomic coordinates 17,033,863–17,035,120; TAIR 10) in Arabidopsis chromosome 4, directs two divergent At4g35985 and At4g35987 genes arranged in head-to-head orientation. (B–C) Schematic diagram of bidirectional promoter-reporter gene constructs: GFP::P85–P87::GUS and GFP::P87–P85::GUS for transient assay in tobacco protoplasts using pB-GFP::P85-P87::GUS and pB-GFP::P87–P85::GUS and for transient Agro-infiltration experiments using pK-GFP::P85–P87::GUS (GeneBank Accession no. KF661330) and pK-GFP::P87–P85::GUS. Below each construct a representative assay of transient GFP expression detected in fluorescence imaging of tobacco protoplast (bar represents 100 µm) and transient GUS expression detected histochemically by Agrobacterium infiltration assay in N. benthamiana leaf are shown for respective promoter At4g35985 promoter (P85) and At4g359857 promoter (P87) activities. Genetic elements: left and right T-DNA border (LT and RT, respectively), CaMV 35S terminator (35ST), green fluorescence protein gene (GFP), β-glucuronidase (GUS), 3′-terminator sequences of ribulose bisphosphate carboxylase small subunit (rbcST) and restriction enzymes EcoRI, HindIII, XhoI, SacI, XbaI, ClaI used to assemble these expression constructs are shown.