Team:Warsaw/Calendar-Main/11 August 2008
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[photo of the gel is to be placed here] | [photo of the gel is to be placed here] | ||
<p>We didn't observe differences in espression 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> | <p>We didn't observe differences in espression 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> | ||
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+ | <h3>Purification of proteins: Z-alpha and Z-omega</h3><h4>Emilia</h4> | ||
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+ | <p><ol><li>Bacteria centrifuged and sonicated (according to <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#Z_a_Z_o>protocol</a>).</li> | ||
+ | <li>Sonicate centrifuged. </li> | ||
+ | <li>Supernatant discarded, pellet resuspended in ddH20, centrifuged and again resuspended in ddH20.</li> | ||
+ | </ol></p> | ||
+ | <br> | ||
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Revision as of 21:14, 19 October 2008
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_A_alpha (with and without induction) and OmpA_A_alpha.
We didn't observe differences in espression 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-omegaEmilia
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