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Team:Warsaw/Calendar-Main/6 August 2008
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| - | Checking if degradation of fusion with OmpA is a result of the activity of lon and | + | Checking if degradation of fusion with OmpA is a result of the activity of lon and ompt proteases (present in <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a>)</h3> |
<h4>Piotr</h4> | <h4>Piotr</h4> | ||
<p>Test was conducted in <i>E.coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a> strain expressing omp_omega_A_alfa (with and without induction) and omp_A_alfa. </p> | <p>Test was conducted in <i>E.coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a> strain expressing omp_omega_A_alfa (with and without induction) and omp_A_alfa. </p> | ||
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Checking if OmpA_omega_A_alpha gives ampicillin resistancePiotrInoculation 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 ompt proteases (present in Top10)PiotrTest 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 |
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