A 1924 bp MtGSTF7 upstream promoter sequence was cloned into the pGreenII 0800-LUC vector between the PstI and NcoI restriction enzyme sites to create the reporter vector proMtGSTF7::LUC. After sequence confirmation, proMtGSTF7::LUC was transformed into A. tumefaciens strain GV3101 that contained the pSoup helper plasmid. The pro35S::cLAP1 and the empty vector pro35S::GUS (pCAMBIA3301) were used as the effector vectors. The A. tumefaciens reporter and effector strains were mixed at a ratio to 1:9 to infiltrate into N. benthamiana leaves, as described previously (Hellens et al., 2005 (link)). The luciferase activity was detected 2 d after infiltration. The primers sequences are listed in Supplementary Table S6 .
To analyse the luciferase activity, 1 mM D-luciferin, potassium salt (D8390, Solarbio, China) was evenly sprayed on the surface of N. benthamiana leaves. After reaction in the dark for 5–10 min, the fluorescence was captured. For quantitative detection analysis, the activities of firefly and Renilla luciferase were quantified using the Dual-Luciferase® Reporter Assay System (E1910, Promega, USA) following the manufacturer’s instructions, and luminescence values were measured using a multifunctional microplate reader (SpectraMax iD3, Molecular Devices, USA).
To analyse the luciferase activity, 1 mM D-luciferin, potassium salt (D8390, Solarbio, China) was evenly sprayed on the surface of N. benthamiana leaves. After reaction in the dark for 5–10 min, the fluorescence was captured. For quantitative detection analysis, the activities of firefly and Renilla luciferase were quantified using the Dual-Luciferase® Reporter Assay System (E1910, Promega, USA) following the manufacturer’s instructions, and luminescence values were measured using a multifunctional microplate reader (SpectraMax iD3, Molecular Devices, USA).
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