diff --git a/studies/1_TagIntegration/README.md b/studies/1_TagIntegration/README.md new file mode 100644 index 0000000000000000000000000000000000000000..20014f90ce6faa2e8df6b785d3199ddca2609ca7 --- /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 Binary files /dev/null and b/studies/1_TagIntegration/isa.study.xlsx differ diff --git a/studies/1_TagIntegration/protocols/.gitkeep b/studies/1_TagIntegration/protocols/.gitkeep new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/studies/1_TagIntegration/protocols/Tag integration.md b/studies/1_TagIntegration/protocols/Tag integration.md new file mode 100644 index 0000000000000000000000000000000000000000..8fda6f713853ea252669fa5699c43d688dea1d59 --- /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 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/studies/1_TagIntegration/resources/20201202_Tagging vectors dtxR.xlsx b/studies/1_TagIntegration/resources/20201202_Tagging vectors dtxR.xlsx new file mode 100644 index 0000000000000000000000000000000000000000..c5d26765d3d2ddccb34ca9e3f1c0d9a9e768e894 Binary files /dev/null and b/studies/1_TagIntegration/resources/20201202_Tagging vectors dtxR.xlsx differ diff --git a/studies/1_TagIntegration/resources/DtxR-His-C-2link.cm5 b/studies/1_TagIntegration/resources/DtxR-His-C-2link.cm5 new file mode 100644 index 0000000000000000000000000000000000000000..5a2118be03c2159dacc22f9befeb5fa65705ffb2 Binary files /dev/null and b/studies/1_TagIntegration/resources/DtxR-His-C-2link.cm5 differ diff --git a/studies/1_TagIntegration/resources/DtxR-Strep-N-4link.cm5 b/studies/1_TagIntegration/resources/DtxR-Strep-N-4link.cm5 new file mode 100644 index 0000000000000000000000000000000000000000..93ef561fccba9ebd17b1bcaaec737c290780701c Binary files /dev/null and b/studies/1_TagIntegration/resources/DtxR-Strep-N-4link.cm5 differ