71340 Sigma-AldrichpCDFDuet™-1 DNA - Novagen

LaneM: Trail Mix™ Protein Markers; Lane1: pACYDuet-1:β-gal + Fluc; Lane2: pETDuet-1:GST/GUS + GFP; Lane3: pRSFDuet-1:Nus/hlFNγ + S•Tag/T4 PNK; Lane4: pCDFDuet-1:GUS + His•Tag/MBP; Lane5: pACYDuet-1, pETDuet-1 combination; Lane6: pACYDuet-1, pRSFDuet-1 combination; Lane7: pACYDuet-1, pCDFDuet-1 combination; Lane8: pETDuet-1, pRSFDuet-1 combination; Lane9: pRSFDuet-1, pCDFDuet-1 combination; Lane10: pETDuet-1, pCDFDuet-1 combination; Lane11: pACYDuet-1, pETDuet-1, pRSFDuet-1 combination; Lane12: pACYDuet-1, pETDuet-1, pCDFDuet-1 combination; Lane13: pACYDuet-1, pRSFDuet-1, pCDFDuet-1 combination; Lane14: pETDuet-1, pRSFDuet-1, pCDFDuet-1 combination; Lane15: Uninduced control; Lane16: All four Duet vectors: all eight proteins

The indicated constructs were transformed individually or together into BL21(DE3). Cultures were grown in TB plus phosphates plus glucose at 37°C to and Abs₆₀₀ between 1.0 and 1.2. Target protein expression was induced by adding IPTG to a final concentration of 1 mM. Cultures were harvested by centrifugation 2.5 h after induction. Lysates were produced by sonication using equal volumes of 1% SDS and 2X SDS sample buffer. Equivalent amounts of protein (based on harvest ABS) were analyzed by SDS-PAGE (4-20% gradient gel) and stained with coomassie blue. To maximize band separation, proteins smaller than 25 kDa were allowed to migrate off the gel.