Talin constructs were synthesized by PCR, using a mouse talin1 cDNA as a template, and cloned into pET-151/D-TOPO (Invitrogen). Constructs were expressed in Escherichia coli (DE3; BL21 Star; Thermo Fisher Scientific) cultured in lysogeny broth (LB). Recombinant His-tagged talin polypeptides were purified as described previously (Goult et al., 2009 (link)). Protein concentrations were determined using the respective extinction coefficients at 280 nm based on absorption coefficients calculated from the aromatic content according to ProtParam.
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Recombinant Talin Polypeptide Purification
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Corresponding Organization :
Other organizations : Yale University, Cancer Research UK Scotland Institute, University of Kent, University of California, San Diego
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Variable analysis
independent variables
- Talin constructs
- Recombinant His-tagged talin polypeptides
- Mouse talin1 cDNA as a template
- PET-151/D-TOPO (Invitrogen) as the cloning vector
- Escherichia coli (DE3; BL21 Star; Thermo Fisher Scientific) as the expression host
- Lysogeny broth (LB) as the culture medium
- Protein concentrations determined using the respective extinction coefficients at 280 nm based on absorption coefficients calculated from the aromatic content according to ProtParam
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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