Fig. 4
afp mRNA and protein expression analysis throughout a time-course of perfusion cultivation. a afp mRNA levels, pictured in diamonds, were measured using qRT-PCR throughout a time period of perfusion cultivation from strain FW23.7 (Panafp::afp) and FW6.6 (PhfbD::afp). Reported standard deviation resulted from a minimum of two and maximum of three measurements per sample. These data support temporal expression profiles reported for mluc gene, with afp expression highest at day 5 for both Panafp and PhfbD. As a control the housekeeping gene h2B was measured in strain PhfbD::afp, which demonstrated a decrease in relative transcript abundance over time (closed circles). b Protein abundance was quantified using Western blot (anti-AFP primary antibody) which demonstrated that Panafp produced maximal 0.06 mgAFP/gdry weight corresponding to 0.65 mgAFP/l. Standard deviation was estimated through three Western blots per sample