From b9d926a2d53addb200356925af41fbd63339fa1b Mon Sep 17 00:00:00 2001
From: Sabrina Zander <sabrina.zander@uni-duesseldorf.de>
Date: Tue, 21 Jan 2025 09:24:56 +0100
Subject: [PATCH] add study1_tag integration

---
 studies/1_TagIntegration/README.md              |   1 +
 studies/1_TagIntegration/isa.study.xlsx         | Bin 0 -> 10594 bytes
 studies/1_TagIntegration/protocols/.gitkeep     |   0
 .../protocols/Tag integration.md                |   2 ++
 studies/1_TagIntegration/resources/.gitkeep     |   0
 .../20201202_Tagging vectors dtxR.xlsx          | Bin 0 -> 20117 bytes
 .../resources/DtxR-His-C-2link.cm5              | Bin 0 -> 4204 bytes
 .../resources/DtxR-Strep-N-4link.cm5            | Bin 0 -> 3578 bytes
 8 files changed, 3 insertions(+)
 create mode 100644 studies/1_TagIntegration/README.md
 create mode 100644 studies/1_TagIntegration/isa.study.xlsx
 create mode 100644 studies/1_TagIntegration/protocols/.gitkeep
 create mode 100644 studies/1_TagIntegration/protocols/Tag integration.md
 create mode 100644 studies/1_TagIntegration/resources/.gitkeep
 create mode 100644 studies/1_TagIntegration/resources/20201202_Tagging vectors dtxR.xlsx
 create mode 100644 studies/1_TagIntegration/resources/DtxR-His-C-2link.cm5
 create mode 100644 studies/1_TagIntegration/resources/DtxR-Strep-N-4link.cm5

diff --git a/studies/1_TagIntegration/README.md b/studies/1_TagIntegration/README.md
new file mode 100644
index 0000000..20014f9
--- /dev/null
+++ b/studies/1_TagIntegration/README.md
@@ -0,0 +1 @@
+Every possible combination was constructed (N-terminal, C-terminal, His, Strep, long linker (4x), short linker (2x)) in every possible combination in pK19-plasmids. However, only two promising were integrated on a first trial (N-Strep-4linker and C-His-2linker), leading to promising strain.
\ No newline at end of file
diff --git a/studies/1_TagIntegration/isa.study.xlsx b/studies/1_TagIntegration/isa.study.xlsx
new file mode 100644
index 0000000000000000000000000000000000000000..ab3d0ac80e56fd5371731bc8e6340dca35ef9c9a
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diff --git a/studies/1_TagIntegration/protocols/.gitkeep b/studies/1_TagIntegration/protocols/.gitkeep
new file mode 100644
index 0000000..e69de29
diff --git a/studies/1_TagIntegration/protocols/Tag integration.md b/studies/1_TagIntegration/protocols/Tag integration.md
new file mode 100644
index 0000000..8fda6f7
--- /dev/null
+++ b/studies/1_TagIntegration/protocols/Tag integration.md	
@@ -0,0 +1,2 @@
+Plasmids were constructed by enzymatically assembling the generated DNA fragments into a cut vector backbone using Gibson assembly (Gibson et al., 2009). Sequencing of the final plasmid was performed by Eurofins Genomics (Ebersberg, Germany).
+For the integration of the His-Tag and a linker (GGGS2) at the C- or N-terminus of DtxR in the genome of C. glutamicum, the suicide vector pK19-mobsacB was used (Schäfer et al., 1994). Electrocompetent C. glutamicum cells were transformed with the isolated plasmid by electroporation (van der Rest et al., 1999). Then, the first and second recombination events were performed and verified as described in previous studies (Niebisch and Bott, 2001). The respective deletion was reviewed by amplification and sequencing (Eurofins Genomics, Ebersberg, Germany).
diff --git a/studies/1_TagIntegration/resources/.gitkeep b/studies/1_TagIntegration/resources/.gitkeep
new file mode 100644
index 0000000..e69de29
diff --git a/studies/1_TagIntegration/resources/20201202_Tagging vectors dtxR.xlsx b/studies/1_TagIntegration/resources/20201202_Tagging vectors dtxR.xlsx
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zr}=35k+@9!nfNC0mlp33_nH4r{DJr<@dcXsH*rejo2-08JVtN+{d36=yR&|5{R38x
BhNl1k

literal 0
HcmV?d00001

-- 
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