Team:ULB-Brussels/Project/Results

From 2014.igem.org

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<h1>Activity of the alkaline phosopahatase with a proline on its N-terminal extremity</h1>
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In order to test the functionality of the alcaline phospatase (phoA) with a proline (P) on its N-terminal extremity, we constructed by restriction processes different plasmids:
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pbad33:: phoA+p (on top), pbad33:: phoA (in the middle)  and we used pbad33 as a control (in the bottom).
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Then we chemoporated the plasmids into a lacking phoA strain and spread the bacteria on different chromogenic culture media including media with glucose (A.2), with arabinose (B.2), and without glucose nor arabinose (A.1 & B.1) as shown in the figure below.</p>
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<h1>Characterization of our biobrick ccdB</h1>
<h1>Characterization of our biobrick ccdB</h1>

Revision as of 14:05, 9 October 2014

$~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ \newcommand{\MyColi}{{\small Mighty\hspace{0.12cm}Coli}} \newcommand{\Stabi}{\small Stabi}$ $\newcommand{\EColi}{\small E.coli} \newcommand{\SCere}{\small S.cerevisae}\\[0cm] ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ \newcommand{\PI}{\small PI}$ $\newcommand{\Igo}{\Large\mathcal{I}} \newcommand{\Tgo}{\Large\mathcal{T}} \newcommand{\Ogo}{\Large\mathcal{O}} ~$ Example of a hierarchical menu in CSS

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- Université Libre de Bruxelles -


Results


Retrieved from "http://2014.igem.org/Team:ULB-Brussels/Project/Results"

Activity of the alkaline phosopahatase with a proline on its N-terminal extremity

In order to test the functionality of the alcaline phospatase (phoA) with a proline (P) on its N-terminal extremity, we constructed by restriction processes different plasmids: pbad33:: phoA+p (on top), pbad33:: phoA (in the middle) and we used pbad33 as a control (in the bottom). Then we chemoporated the plasmids into a lacking phoA strain and spread the bacteria on different chromogenic culture media including media with glucose (A.2), with arabinose (B.2), and without glucose nor arabinose (A.1 & B.1) as shown in the figure below.

Characterization of our biobrick ccdB

X. 01/09/14-07/09/14

We finished with a killing ccdB test for the characterization of pSB_1C3-ccdB: this is ok :)

In the two following screens (the first is with presence of glucose, the second with arabinose), the first line corresponds to $\EColi$ without Tox-plasmid and without A-Tox plasmid (no plasmid), the second to $\EColi$ with Tox-plasmid but without A-Tox plasmid (pBAD33::ccdB), the third to $\EColi$ without Tox-plasmid but with A-Tox plasmid (pKK233::ccdA), and the last to $\EColi$ with the two plasmids (pKK233::ccdA & pBAD33::ccdB). The columns vary by a dilution factor (from left to right: $10^{0}$, $10^{-2}$, $10^{-3}$, $10^{-4}$, $10^{-6}$).

$E.Coli \hspace{0.2cm}+\hspace{0.2cm} glucose:\hspace{1cm}$

$E.Coli \hspace{0.2cm}+\hspace{0.2cm} arabinose:\hspace{0.65cm}$

With these two killing assay screens, we see that the glucose represses the toxin (line two: bacteria survive)

and arabinose inducts the production of toxin, and bacteria death.
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