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10.29219/fnr.v62.1415 71 LeonardiR.JackowskiS

J personalized Med

Gibco), 5 mM glutamine and 5 mM Pen Strep, and sub-cultured regularly

Abstract Background Epidemiologic evidence, reinforced by clinical and laboratory studies, shows that the rich Western diet is the major underlying cause of death and disability (e.g, from cardiovascular disease and type 2 diabetes) in Western industrialized societies

Additionally, the Nrf2/NF-B pathway was regulated to eliminate ROS

N-Terminal Fusion (Default) Advantages: Fusion partner folds first (translated first), creating a folding nucleus Most protease cleavage sites are designed for N-terminal fusions Better studied, more reliable data Disadvantages: Blocks the native N-terminus If the N-terminus is functionally important, the fusion may interfere C-Terminal Fusion Advantages: Preserves native N-terminus Some proteins fold better with a free N-terminus (Sachdev & Bhatt, 2011) Can stabilize proteins that are prone to C-terminal degradation Disadvantages: Fusion partner folds after the passenger (less folding nucleation effect) Fewer validated cleavage strategies Read-through or premature termination can produce truncated fusions When to Try C-Terminal N-terminal fusion tested and failed N-terminus is known to be important for function Protein has a signal peptide or pro-peptide at the N-terminus Cleavage Strategies Protease Comparison Recommendation: TEV or Ulp1 for most applications