Plasmid: available using <i>LabGenius BioBrick Mapper</i> <br>
Plasmid: available using <i>LabGenius BioBrick Mapper</i> <br>
Revision as of 07:34, 12 October 2014
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Name: BBa_K1318000
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].
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 killing 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's expression), each colony grew. That allowed us to control the non toxicity of ccdA.
On the media containing both IPTG and arabinose (inducing the pBAD's expression), the strand with pBAD was killed and the strand with both ccdA $\small\&$ ccdB grew.
We made dilutions to assure that the cell concentration didn’t affect the toxicity or the anti-toxicity.
"
- Université Libre de Bruxelles -
