Selected article for: "capping activity and luciferase expression"

Author: Jaïs, Philippe H; Decroly, Etienne; Jacquet, Eric; Le Boulch, Marine; Jaïs, Aurélien; Jean-Jean, Olivier; Eaton, Heather; Ponien, Prishila; Verdier, Fréderique; Canard, Bruno; Goncalves, Sergio; Chiron, Stéphane; Le Gall, Maude; Mayeux, Patrick; Shmulevitz, Maya
Title: C3P3-G1: first generation of a eukaryotic artificial cytoplasmic expression system
  • Document date: 2019_3_18
  • ID: 6nq7y1qe_55
    Snippet: As an alternative to leucine-zippers, we tested the functional activity of fusing vaccinia virus capping enzyme subunits to T7RNAP. The D1R or D12L ORFs were fused in-frame to the amino-terminus of T7RNAP, separated by a flexible (G 4 S) 4 linker, generating D1R-(G 4 S) 4 -T7RNAP and D12L-(G 4 S) 4 -T7RNAP constructs respectively. In comparison to uncoupled enzymes, fusion increased luciferase expression by 8-and 3.9-fold respectively ( Figure 2 .....
    Document: As an alternative to leucine-zippers, we tested the functional activity of fusing vaccinia virus capping enzyme subunits to T7RNAP. The D1R or D12L ORFs were fused in-frame to the amino-terminus of T7RNAP, separated by a flexible (G 4 S) 4 linker, generating D1R-(G 4 S) 4 -T7RNAP and D12L-(G 4 S) 4 -T7RNAP constructs respectively. In comparison to uncoupled enzymes, fusion increased luciferase expression by 8-and 3.9-fold respectively ( Figure 2 ). In both approaches, coupling of D1R to T7RNAP outperformed coupling of D12L to RNAP and confirmed the importance of physically coupling capping enzyme and RNAP enzymes for T7-transcript expression.

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