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Plasmid DNA Preparation> | … For our RD construct, a gene encoding residues 373–468 of CaN (NCBI NP_000935) was synthesized by Genscript (Piscataway, NJ) and subcloned into a pET303 plasmid (Life Technologies, Grand Island, NY), which adds a C-terminal six-His tag … | Get A Quote |
Calcineurin is a Ser/Thr phosphatase that is important for key biological processes, including immune system activation. We previously identified a region in the intrinsically disordered regulatory domain of calcineurin that forms a critical amphipathic α-helix (the "distal helix") that is required for complete activation of calcineurin. This distal helix was shown to have a Tm close to that of human body temperature. Because the Tm was determined in dilute buffer, we hypothesized that other factors inherent to a cellular environment might modulate the stability of the distal helix. One such factor that contributes to stability in other proteins is macromolecular crowding. The cell cytoplasm is comprised o... More