The caspase-6 variants, full length wild type (FL WT), FL C163S and constitutively two-chain (CT) were derived from the synthetic, E. coli codon-optimized (His)6 C-terminally tagged caspase-6 gene (Celtek Bioscience) that were ligated into the NdeI/BamHI sites of pET11a vector.31 (link) Selected caspase-6 tumor-associated variants were generated using FL WT as template through Phusion® site-directed mutagenesis (Thermo Scientific™). The caspase-6 R259H CT and W227A CT variants were particularly generated using caspase-6 D179CT as the template for the site-directed mutagenesis.
Protocol detail
Generating Caspase-6 Variant Proteins
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Other organizations : University of Massachusetts Amherst
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Variable analysis
independent variables
- Caspase-6 variants: full length wild type (FL WT), FL C163S, and constitutively two-chain (CT)
- Not explicitly mentioned
- The caspase-6 gene was derived from a synthetic, E. coli codon-optimized (His)6 C-terminally tagged caspase-6 gene and ligated into the NdeI/BamHI sites of pET11a vector
- The caspase-6 R259H CT and W227A CT variants were generated using caspase-6 D179CT as the template for the site-directed mutagenesis
- No positive or negative controls were explicitly mentioned in the provided information
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