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Team:Warsaw/Calendar-Main/11 August 2008
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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 <A href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>OmpA_omega_ΔA_alpha</a> (with and without induction) and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-deltaA-alpha>OmpA_ΔA_alpha</a>. | 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 <A href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>OmpA_omega_ΔA_alpha</a> (with and without induction) and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-deltaA-alpha>OmpA_ΔA_alpha</a>. | ||
<ol> | <ol> | ||
| - | <li> | + | <li>Spinning.</li> |
<li>Suspending.</li> | <li>Suspending.</li> | ||
| - | <li>Adding | + | <li>Adding the lysis buffer.</li> |
<li>Boiling.</li> | <li>Boiling.</li> | ||
<li>Putting into poliacrylamide gel.</li> | <li>Putting into poliacrylamide gel.</li> | ||
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| - | <p>We didn't observe differences in | + | <p>We didn't observe differences in expression and degradation in <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosettas</a> nor in <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>. Therefore we suppose that degradation of the fusions is caused by factor other than Lon and OmpT proteases.</p> |
Revision as of 23:40, 25 October 2008
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Checking whether degradation of the fusions with OmpA is caused by Lon and OmpT proteases (present in TOP10)PiotrTest was conducted in E.coli Rosetta strain expressing OmpA_omega_ΔA_alpha (with and without induction) and OmpA_ΔA_alpha.
Detection of protein A. Dalszy opis również wstawię jak się wyśpię...
We didn't observe differences in expression and degradation in Rosettas nor in TOP10. Therefore we suppose that degradation of the fusions is caused by factor other than Lon and OmpT proteases. Purification of proteins: Z-alpha and Z-omegaEmilia100 ml of medium inoculated with overnight culture, then used as inoculum for 1 l culture in presence of ampicillin, kanamycin and IPTG (according to protocol).
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