From 21606fd8bb59d5f00f2c3602a23eeea931489cc2 Mon Sep 17 00:00:00 2001
From: Viktoria Petrova <vipet103@hhu.de>
Date: Sat, 9 Nov 2024 15:19:54 +0100
Subject: [PATCH] add construction of gus reporter lines assay and protocol

---
 assays/ConstructionOfGUSReporterLines/README.md  |   0
 .../dataset/.gitkeep                             |   0
 .../isa.assay.xlsx                               | Bin 0 -> 6909 bytes
 .../protocols/.gitkeep                           |   0
 ...onstructionOfTranslationalGUSReporterLines.md |   3 +++
 5 files changed, 3 insertions(+)
 create mode 100644 assays/ConstructionOfGUSReporterLines/README.md
 create mode 100644 assays/ConstructionOfGUSReporterLines/dataset/.gitkeep
 create mode 100644 assays/ConstructionOfGUSReporterLines/isa.assay.xlsx
 create mode 100644 assays/ConstructionOfGUSReporterLines/protocols/.gitkeep
 create mode 100644 assays/ConstructionOfGUSReporterLines/protocols/ConstructionOfTranslationalGUSReporterLines.md

diff --git a/assays/ConstructionOfGUSReporterLines/README.md b/assays/ConstructionOfGUSReporterLines/README.md
new file mode 100644
index 0000000..e69de29
diff --git a/assays/ConstructionOfGUSReporterLines/dataset/.gitkeep b/assays/ConstructionOfGUSReporterLines/dataset/.gitkeep
new file mode 100644
index 0000000..e69de29
diff --git a/assays/ConstructionOfGUSReporterLines/isa.assay.xlsx b/assays/ConstructionOfGUSReporterLines/isa.assay.xlsx
new file mode 100644
index 0000000000000000000000000000000000000000..25d8c928d77c1ea574abfe9a6c56589aa1192308
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diff --git a/assays/ConstructionOfGUSReporterLines/protocols/.gitkeep b/assays/ConstructionOfGUSReporterLines/protocols/.gitkeep
new file mode 100644
index 0000000..e69de29
diff --git a/assays/ConstructionOfGUSReporterLines/protocols/ConstructionOfTranslationalGUSReporterLines.md b/assays/ConstructionOfGUSReporterLines/protocols/ConstructionOfTranslationalGUSReporterLines.md
new file mode 100644
index 0000000..a5b632e
--- /dev/null
+++ 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
-- 
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