Fig. 1
Design for the engineering of recombinant DNA for subsequent insertion into the pGEX-6P1 plasmid. A Schematic representation of the cytosolic tail of the human CB1 receptor showing its primary amino acid sequence. The helix 8 and helix 9 structural domains are highlighted in violet and cyan, respectively. The arrows indicate the initial and terminal amino acids of the polypeptides ranging from serine 414 to terminal lysine 472 (CB1414–472) and from serine 414 to valine 442 (CB1414–442) chosen for the design of the fusion proteins GST-CB1414–472 and GST-CB1414–442. Illustration modified from Stadel and Kendall [64]. B DNA products obtained by PCR amplification and the encoded polypeptides CB1414–472 and CB1414–442. The underlined DNA sequences indicate the hybridization sites for the Fw and Rv primers. The BamHI and NotI restriction sites for cloning into the pGEX-6P1 vector are highlighted in green and yellow, respectively. The triplets encoding serine 414 in the human CB1 receptor and the stop codons are highlighted in cyan and red, respectively