Team:UNICAMP-Brazil/Notebooks/October 15
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==== Testing the new device ==== | ==== Testing the new device ==== | ||
- | *<p style=”text-align:justify;”>We confirmed our device yesterday, but we continued working with it. We proved that our device really works. To fulfill this aim, we transformed competent E. Coli C43. This E. coli strain can | + | *<p style=”text-align:justify;”>We confirmed our device yesterday, but we continued working with it. We proved that our device really works. To fulfill this aim, we transformed competent ''E. Coli'' C43. This ''E. coli'' strain can produce the T7 polimeraze by IPTG induction. Thus, when we put IPTG into the culture medium, we will have expression of T7 polimeraze, which will induce the T7 promoter of our device and then, the endolysin will be expressed. Our test consists in overexpression of the endolysin in order to break it's own peptidoglycan wall. We transformed cells with our device BBa_K284022 plasmid and with the BBa_K112806 plasmid (without promoter).</p> |
- | *<p style=”text-align:justify;”>So, after get the transformed | + | *<p style=”text-align:justify;”>So, after get the transformed cells, we put them to growth in 5 erlenmeyers (LB-AMP medium) until half log phase (OD-0.6 to 1). Then, we induced with IPTG (1mM) and measured the optical density during 4 hours after induction. We plated the 5 cultures in LB-AMP medium plates in order to confirm the diminished cellular growth. To have a definitive confirmation, we also performed SDS-PAGE to notice the protein overexpression. The results are shown in [https://2009.igem.org/Team:UNICAMP-Brazil/Coliguard/Results ColiGuard results].</p> |
''Luige, Ane & Marcos'' | ''Luige, Ane & Marcos'' |
Revision as of 02:54, 22 October 2009
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