Team:UNICAMP-Brazil/Notebooks/October 15

(Difference between revisions)

Revision as of 00:18, 22 October 2009

Fabi and Léo

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 be producing 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 so, the endolysin will be expressed. Our test consists in superexpress the endolysin in order to break itself peptidoglycan wall. We transformed cells with our device BBa_K284022 plasmid and with the BBa_K112806 plasmid (without promoter).
So, after get the transformed cell, we put them to growth in 5 erlenmeyers (ampicilim LB medium) until half log phase(OD-0.6 to 1). Then, we induced with IPTG (1mM) and we measured the optical density for 4 hours after induction. We plated the 5 cultures in ampicilin LB medium plates in order to confirm the diminished cellular growth. To have a definitive confirmation, we also performed PAGE-SDS to notice the protein superexpression. We show the result in ColiGuard results.

Luige, Ane & Marcos

We sent 4 biobricks to iGEM today: pJEN1, pDLD, Lysozyme, and the devices Adh1+YFP and Adh1+Lysozyme. =)

We transformed the YEP+Adh1-lysozyme ligation reaction in competent E. coli and plated in LB+Amp media. Hope to have colonies later!

We did miniprep of YEP+Adh1-YFP and the following devices: b) pJEN1+YFP, c) pJEN1+Lys, d) pDLD+YFP, e) pDLD+Lys.

Raíssa and Taís

@@ Line 12: / Line 12: @@
 ''Fabi and Léo''
+==== 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 be producing 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 so, the endolysin will be expressed.   Our test consists in superexpress the endolysin in order to break itself 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 cell, we put them to growth in 5 erlenmeyers (ampicilim LB medium) until half log phase(OD-0.6 to 1). Then, we induced with IPTG (1mM) and we measured the optical density for 4 hours after induction. We plated the 5 cultures in ampicilin LB medium plates in order to confirm the diminished cellular growth.  To have a definitive confirmation, we also performed PAGE-SDS to notice the protein superexpression.  We show the result in [https://2009.igem.org/Team:UNICAMP-Brazil/Coliguard/Results ColiGuard results].</p>
+''Luige, Ane & Marcos''
 ==''' YeastGuard '''==