Construct σ1long, comprising the three most C-terminal β-spirals of T1L σ1 and the entire head domain (amino acids 261 to 470), was purified as described previously (22 (link)) and used in STD-NMR experiments. Mutations were generated using the GeneArt site-directed mutagenesis PLUS system (Invitrogen) along with AccuPrime Pfx DNA polymerase (Invitrogen) and One Shot MAX Efficiency DH5α-T1R competent cells (Invitrogen).
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Purification and Mutagenesis of σ1 Protein
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Corresponding Organization :
Other organizations : Vanderbilt University, University of Tübingen, Texas Children's Hospital, Baylor College of Medicine, Children's Cancer Center
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Variable analysis
independent variables
- Mutations generated using the GeneArt site-directed mutagenesis PLUS system (Invitrogen)
- Outcomes of the STD-NMR experiments
- Amino acids 261 to 470 of T1L σ1, comprising the three most C-terminal β-spirals and the entire head domain, purified as described previously (22)
- Positive control: Not specified
- Negative control: Not specified
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