Investigating the molecular mechanism of positive and negative allosteric modulators in the calcium-sensing receptor dimer
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The previously described myc-tagged rat GPRC6A-pEGFPN1 construct, which contains the mGluR5 signal peptide upstream of the myc tag51, was used as template for generating the CaSR constructs used in the present study. myc-tagged CaSR was generated by replacing the rat GPRC6A sequence with the rat CaSR sequence by use of the restriction sites MluI and NotI. A sequence comprised of the mGluR5 signal peptide and an HA tag was synthesized by Genscript (Piscataway, NJ, USA) and inserted in myc-CaSR-pEGFPN1 instead of the mGluR5 signal peptide and the myc tag by use of XhoI and MluI thus generating HA-CaSR. |
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