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+++ b/assays/ConstructionOfGUSReporterLines/protocols/ConstructionOfTranslationalGUSReporterLines.md @@ -0,0 +1,3 @@ +## Construction of translational GUS reporter lines + +Genomic fragments comprising the 5′-upstream region and the entire coding region of *SWEET1, SWEET4, SWEET5, SWEET7, SWEET8, SWEET10,* and *SWEET13* were amplified by PCR using wild-type *A. thaliana* Col-0 genomic DNA as a template (primers for each gene are listed in Key Resource Table). The amplified products of *SWEET4, SWEET5, SWEET7, SWEET8*, and *SWEET10* were cloned into promoterless GUSplus vector (Yang, J. et al., 2018) while *SWEET1* and *SWEET13* were cloned into pMDC163 using the Gateway cloning system. All constructs were confirmed by sequencing. Wild-type *A. thaliana* Col-0 were transformed by *Agrobacterium tumefaciens* harboring SWEET-GUS constructs using the floral dip method. Transgenic seedlings were selected on ½ MS media supplemented with 1% sucrose and 25 μg/mL hygromycin. Transgenic *SWEET2-GUS, SWEET11-GUS, SWEET12-GUS* and *SWEET17-GUS* were previously reported in Chen et al. (2012), Chen et al. (2015), and Guo et al. (2014). \ No newline at end of file -- GitLab