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Fig. 6 | BMC Plant Biology

Fig. 6

From: The role of protein-protein interactions mediated by the PB1 domain of NLP transcription factors in nitrate-inducible gene expression

Fig. 6

Activity of mutated NLP7 in a protoplast transient assay system. a Schematic representation of the reporter constructs used in b and c; LUC: firefly luciferase gene; NOS: transcription termination sequence of the nopaline synthase gene. White boxes indicate 5′ or 3′ untranslated regions, and horizontal lines indicate sequences upstream (“promoter”) or downstream of the NRT2.1, NIR1, or NIA1 coding regions. Green ovals mark experimentally verified NLP-binding sites. b Protoplast transient assay using N-starved Col protoplasts. Protoplasts were co-transfected with the NRT2.1pro reporter plasmid, effector plasmids for expression of wild-type or mutated NLP7, and a control plasmid expressing β-glucuronidase (GUS) under the control of the UBQ10 promoter (UBQ10-GUS) and incubated overnight in medium supplemented with either 1 mM KCl or KNO3. c Protoplast transient assay using protoplasts isolated from leaves of nlp6 nlp7–1 plants supplied with nitrogen. Protoplasts were co-transfected with the reporter plasmid, plasmids for expression of NLP7, and the control plasmid, and incubated overnight in protoplast incubation medium. Luciferase activity values were normalized against GUS activity. Means ± SD (n = 3) are shown in b and c

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