Fig 5. CFPS using a linear DNA template and LoFT cell extract.

sfGFP was produced using LoFT cell extract and 5 nM linear DNA (PCR product) for 1 h at 29°C, in the absence (−) or presence (+) of 4 μM GamS. OR and T7 indicate PCR products from the OR2OR1-sfGFP (from the pOR2OR1-sfGFP plasmid) and T7-sfGFP (from the pET29-sfGFP plasmid), respectively. Prom. means “Promoter”. (A) Expression levels of GFP estimated by total fluorescence levels. Error bars indicate standard deviations (n = 3). (B) GFP fluorescence detected using SDS-PAGE without boiling. Only bands corresponding to GFP fluorescence are shown.