Team:Warsaw/Calendar-Main/6 August 2008

Checking if OmpA_omega_&Delta;A_alpha gives ampicillin resistance Piotr, Weronika

Repetition of the test from previous day. Inoculation of OmpA_omega_&Delta;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 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_&Delta;A_alpha (with and without induction) and ompA_A_alpha.

Transformation of Rosettas with OmpA_omega_&Delta;A_alpha and OmpA_A_alpha</a>.

Preparing pACYC177+OmpA_omega_&Delta;A</a> construct Michał K.  Inoculation of cultures from colonies grown on plate with pACYC177+OmpA_omega_&Delta;A</a> transformation.