Checking if OmpA_omega_ΔA_alpha gives ampicillin resistance

Piotr

Inoculation of OmpA_omega_ΔA_alpha from various IPTG concentrations: 0, 0.1, 0.25, 0.5, 0.75, 1 mM into same IPTG concentrations, but with various ampicillin concentrations (25, 50, 75, 100 mM) in ratio 1:50.

Conclusion: we obtained the best induction using 0.5 - 0.25 mM IPTG. Best ampicillin concentration is 50 - 75 μg/ml.

Checking whether degradation of the fusions with OmpA is caused by Lon and OmpT proteases (present in TOP10)

Emilia

Test was conducted in E.coli Rosetta strain expressing ompA_omega_ΔA_alpha (with and without induction) and ompA_A_alpha.

Transformation of Rosettas with OmpA_omega_ΔA_alpha and OmpA_A_alpha.

Preparing pACYC177+OmpA_omega_ΔA construct

Michał K.

Inoculation of cultures from colonies grown on plate with pACYC177+OmpA_omega_ΔA transformation.