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Fig. 3 | BMC Biochemistry

Fig. 3

From: Soluble expression of recombinant midgut zymogen (native propeptide) proteases from the Aedes aegypti Mosquito Utilizing E. coli as a host

Fig. 3

Soluble expression of recombinant AaET-NL and AaSPVII-NL zymogen proteases grown in TB media at 23 °C post-induction (induced with 0.1 mM IPTG). Plasmid constructs were transformed into SHuffle® T7 Express Competent E. coli cells (NEB). The MW ladder is in kilo-Daltons (kDa). a Western blot analysis utilizing an AaET-specific antibody of soluble samples collected from the growth and expression of AaET (a total of 4 h post-induction). The zymogen (inactive form of the protease) is observed in the first 2 h (MW ~ 27.0 kDa, red arrow), but a second species hypothesized to be the active mature form begins to appear at the two-hour time-point (MW ~ 22.4 kDa, green arrow) while the zymogen completely disappears by the third hour post-induction. b Large scale expression analysis of AaSPVII-zymogen grown for a total of 5 h post-induction. A single band at ~ 28.7 kDa (orange arrow) is observed to be increasing over time after induction with no observable band present in both the total and soluble pre-induction samples (t = 0 h)

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BMC Biochemistry

ISSN: 1471-2091

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