RHEB (NM_005614) Human Tagged ORF Clone

Snippet: … A plasmid carrying Rheb was purchased from OriGene Technologies (Rockville, MD, USA). Rheb DNA was amplified and modified to incorporate a FLAG-encoding sequence at the 3'-end by expanded long-template PCR (Roche, Basel, Switzerland). Rheb(S16H) was generated with the Phusion Site-Directed Mutagenesis Kit (New England Biolabs, Ipswich, MA, USA) in the pGEM-T vector (Promega, Madison, WI, USA), and then cloned into an AAV packaging construct that utilizes the chicken β-actin promoter and contains a 3' WPRE (pBL). AAVs were produced by the University of North Carolina Vector Core, and the genomic titer of Rheb(S16H) was 2×1012 viral genomes/ml. …

Snippet: The experimental scheme is provided in Figure 1A. 2.2. Production of AAV Viral Vectors. All vectors used were AAV1 serotype as previously described [15,16]. A plasmid carrying Rheb was purchased from OriGene Technologies (Rockville, MD, USA) Cached

Snippet: All vectors used were the AAV1 serotype as previously described (Kim & Chen et al., 2011;Kim & Kareva et al., 2012). A plasmid carrying Rheb was purchased from OriGene Technologies (Rockville, MD, USA). Rheb DNA was amplified and modified to incorporate

Snippet: The MYR-HA-hAKT3 sequence was amplified and cloned into an AAV proviral expression plasmid using the In-Fusion HD cloning system (Clontech, Mountain View, CA, USA). The human Rheb cDNA clone was obtained from Origene (Rockville, MD, USA)