Human Galpha-s Expression

The goal of this subproject is to express human G-alpha-s subunit in Xenopus oocytes together with TAAR1 to facilitate its coupling to GIRK channels. We purchased pcDNA3.1.Gαs-LgB113 (Plasmid #134362) from addgene.

The gene that codes for the Guanine nucleotide-binding protein G(s) subunit alpha isoforms short protein (Gαs) is called Human GNAS2. 

It has the following sequence: MGCLGNSKTEDQRNEEKAQREANKKIEKQLQKDKQVYRATHRLLLLGAGESGKSTIVKQMRILHVNGFNGEGGEEDPQAARSNSDGEKATKVQDIKNNLKEAIETIVAAMSNLVPPVELANPENQFRVDYILSVMNVPDFDFPPEFYEHAKALWEDEGVRACYERSNEYQLIDCAQYFLDKIDVIKQADYVPSDQDLLRCRVLTSGIFETKFQVDKVNFHMFDVGGQRDERRKWIQCFNDVTAIIFVVASSSYNMVIREDNQTNRLQEALNLFKSIWNNRWLRTISVILFLNKQDLLAEKVLAGKSKIEDYFPEFARYTTPEDATPEPGEDPRVTRAKYFIRDEFLRISTASGDGRHYCYPHFTCAVDTENIRRVFNDCRDIIQRMHLRQYELL

In the plasmid an additional sequence is inserted between the highlighted amino acids and is as follows: DISGGGGSVFTLEDFVGDWEQTAAYNLDQVLEQGGVSSLLQNLAVSVTPIQRIVRSGENALKIDIHVIIPYEGLSADQMAQIEEVFKVVYPVDDHHFKVILPYGTLVIDGVTPNMLNYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLITPDGSMLFRVTINSSGGGGSPR

This was found to be nanoluciferase (LgB113) inserted between residues 113 and 114 of Gαs.