Team:ULB-Brussels/OurBrick

From 2014.igem.org

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Our MightyColi BioBrick
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The MightyColi BioBrick
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<h1> Our Part </h1>
<h1> Our Part </h1>
<h3> Properties </h3>
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<p> Host: E.Coli <br>
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<p> Host: E.coli <br>
Source: ccdBA operon <br>
Source: ccdBA operon <br>
Plasmid: available using <i>LabGenius BioBrick Mapper</i> <br>
Plasmid: available using <i>LabGenius BioBrick Mapper</i> <br>
Length: 309 bp <br>
Length: 309 bp <br>
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Sequence: (begin) atgcagttt ... atataataa (end) <br>
Compatibility: RFC[10], RFC[12], RFC[21], RFC[23], RFC[25].
Compatibility: RFC[10], RFC[12], RFC[21], RFC[23], RFC[25].
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Revision as of 15:51, 7 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 -



The MightyColi BioBrick


Our Part

Properties

Host: E.coli
Source: ccdBA operon
Plasmid: available using LabGenius BioBrick Mapper
Length: 309 bp
Sequence: (begin) atgcagttt ... atataataa (end)
Compatibility: RFC[10], RFC[12], RFC[21], RFC[23], RFC[25].

$\Longrightarrow$ ULB-Brussels part

Characterization

In order to characterize the ccdB biobrick, we sent the biobricks to sequencing and made a screen of activity for the protein ccdB. We did a $\small killing$ $\small assay$, because of the toxic property of ccdB.

We constructed 4 different colonies including one with the plasmid pKK-233-ccda, another with pBAD33-ccdB, a third with both, and a control colony. The ccdA gene encoded for a protein wich acts as an anti-toxin of ccdB.

On the first media containing IPTG (inducing the pKK233’s expression) and glucose (repressing the pBAD expression), each colony grew. That allowed us to control the non toxicity of ccdA. On the media containing both IPTG and arabinose, the strand with pBAD is killed and the strand with both ccdA and ccdB grew.

We made dilution to assure that the cell concentration didn’t affect the toxicity or the anti-toxicity.

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