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The human TREK-2 (NM_138318)- and TREK-1 (NM_014217) -expressing vector (pGH19-TREK-2) was constructed as described19. On the basis of this vector, point Mutations and deletions were engineered using the MutanBEST kit (TaKaRa, Dalian, China)-guided high-fidelity PCR. All the mutations were confirmed by DNA sequencing. All the Plasmids were linearized by Xho I before in vitro transcription. cRNA was synthesized using the RiboMAX™ Large Scale RNA Production Systems (Promega, Madison, WI) kit.
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