Checking if OmpA_omega_A_alpha gives ampicillin resistance

Piotr

Inoculation of OmpA_omega_A_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 if degradation of fusion with OmpA is a result of the activity of lon and iompt proteases (present in Top10)

Piotr

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

Transformation of Rosettas with omega_A_alpha and A_alpha.

Preparing pACYC177+OmpA_omega_deltaA construct

Michał K.

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