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Team:Warsaw/Stage2/Design
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<h2>Design</h2> | <h2>Design</h2> | ||
<div class="note"> Design and measurement of kill-switch safety system </div> | <div class="note"> Design and measurement of kill-switch safety system </div> | ||
| - | <p>The safety system we designed and measured consists of the parts specified: <a href="http://partsregistry.org/Part:BBa_I719005">T7 promoter</a> + <a href="http://partsregistry.org/Part: | + | <p>The safety system we designed and measured consists of the parts specified: <a href="http://partsregistry.org/Part:BBa_I719005">T7 promoter</a> + <a href="http://partsregistry.org/Part:BBa_B0032">B0032</a> RBS + <a href="http://partsregistry.org/Part:BBa_K299806">MinC</a> on pSB1A2 plasmid, studied in <i>E.coli</i> BL21 RIL strain. </p> |
</html><partinfo>BBa_K299807 DeepComponents</partinfo><html> | </html><partinfo>BBa_K299807 DeepComponents</partinfo><html> | ||
| - | <p>We measured our construct in vivo, in the expression system, using strong T7 promoter, IPTG induced. The functionality of our safety system describes optical density of E.coli (OD) and the number of colony forming units | + | <p>We measured our construct <i>in vivo</i>, in the expression system, using strong <a href="http://partsregistry.org/Part:BBa_I719005">T7 promoter</a>, IPTG induced (see <a href="https://2010.igem.org/Team:Warsaw/Stage2/Results">results</a> section). The functionality of our safety system describes optical density of <i>E.coli</i> cells (OD) and the number of colony forming units per mililiter (cfu/ml) in IPTG-induced innoculates BL21 carrying our construct.</p> |
<div class="note"> Additional experiment to kill-switch safety system </div> | <div class="note"> Additional experiment to kill-switch safety system </div> | ||
| - | <p>We also performed experiment with B0034 RBS as an alternative to B0032. We designed biobrick presented below: | + | <p>We also performed experiment with <a href="http://partsregistry.org/Part:BBa_B0034">B0034</a> RBS as an alternative to <a href="http://partsregistry.org/Part:BBa_B0032">B0032</a>. We designed biobrick presented below: |
| - | <a href="http://partsregistry.org/Part:BBa_I719005">T7 promoter</a> + <a href="http://partsregistry.org/Part: | + | <a href="http://partsregistry.org/Part:BBa_I719005">T7 promoter</a> + <a href="http://partsregistry.org/Part:BBa_B0034">B0034</a> RBS + <a href="http://partsregistry.org/Part:BBa_K299806">MinC</a> on pSB plasmid, studied in E.coli BL21 strain. </p> |
</html><partinfo>BBa_K299808 DeepComponents</partinfo><html> | </html><partinfo>BBa_K299808 DeepComponents</partinfo><html> | ||
| - | As we know from RBS measurement data | + | As we know from RBS <a href="https://2010.igem.org/Team:Warsaw/Stage1/RBSMeas">measurement data</a>, <a href="http://partsregistry.org/Part:BBa_B0034">B0034</a> is stronger than <a href="http://partsregistry.org/Part:BBa_B0032">B0032</a>. As a result, we didn’t obtain any visible bacterial colonies on agar plates (see <a href="https://2010.igem.org/Team:Warsaw/Stage2/Results">results</a>). Our conslucion to this experiment is that additional regulation of <a href="http://partsregistry.org/Part:BBa_K299806">MinC</a> gene expression is very important to keep the kill-switch system working. </p> |
</html> | </html> | ||
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Latest revision as of 18:56, 27 October 2010
Design
Design and measurement of kill-switch safety system
The safety system we designed and measured consists of the parts specified: T7 promoter + B0032 RBS + MinC on pSB1A2 plasmid, studied in E.coli BL21 RIL strain.
<partinfo>BBa_K299807 DeepComponents</partinfo>We measured our construct in vivo, in the expression system, using strong T7 promoter, IPTG induced (see results section). The functionality of our safety system describes optical density of E.coli cells (OD) and the number of colony forming units per mililiter (cfu/ml) in IPTG-induced innoculates BL21 carrying our construct.
Additional experiment to kill-switch safety system
We also performed experiment with B0034 RBS as an alternative to B0032. We designed biobrick presented below: T7 promoter + B0034 RBS + MinC on pSB plasmid, studied in E.coli BL21 strain.
<partinfo>BBa_K299808 DeepComponents</partinfo> As we know from RBS measurement data, B0034 is stronger than B0032. As a result, we didn’t obtain any visible bacterial colonies on agar plates (see results). Our conslucion to this experiment is that additional regulation of MinC gene expression is very important to keep the kill-switch system working.
