NADPH-Cytochrome P450 Reductase: Molecular Cloning and Functional Characterization of Two Paralogs from Withania somnifera (L.) Dunal
Figure 5
Sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) of heterologously expressed WsCPR1 and WsCPR2.
5A: SDS-PAGE (10%) pattern of proteins obtained from E.coli BL21 (DE3) transformed with pGEX-WsCPR1 and pGEX-WsCPR2. Lane 1 & 5; Standard protein markers. Lane 2 & 7; Whole cell lysate of uninduced WsCPR1 and WsCPR2, Lane 3–6 & 10–13; Cell lysate of WsCPR1 and WsCPR2 induced with 0.8 mM IPTG harvested after 2 h, 4 h, 6 h and 8 h respectively. Lane 7 & 14; Cell lysate of E. coli BL21 (DE3) cells containing the empty vector pGEX-4T2 obtained at 4 h post-induction with 0.8 mM IPTG. 5B: The SDS-PAGE (10%) profile of optimized WsCPR2 expression at 25°C. In different lanes the samples were harvested at different time periods as indicated above. The highest expression was observed with 0.8 mM IPTG after 12 h of induction.