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Team:Warsaw/Stage2/Design
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<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 B0034 RBS as an alternative to B0032. 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_K299806">MinC</a> + <a href="http://partsregistry.org/Part:BBa_B0034">RBS B0034</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 (link), B0034 is stronger than B0032. As a result, we didn’t obtain any visible bacterial colonies on agar plates. Our conslucion to this experiment is that additional regulation of MinC gene expression is very important to keep the system working. </p> | As we know from RBS measurement data (link), B0034 is stronger than B0032. As a result, we didn’t obtain any visible bacterial colonies on agar plates. Our conslucion to this experiment is that additional regulation of MinC gene expression is very important to keep the system working. </p> | ||
Revision as of 14:55, 24 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 + MinC + RBS B0032 on pSB plasmid, studied in E.coli BL21 strain.
<partinfo>BBa_K299807 DeepComponents</partinfo>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/ ml (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 + MinC + RBS B0034 on pSB plasmid, studied in E.coli BL21 strain.
<partinfo>BBa_K299808 DeepComponents</partinfo> As we know from RBS measurement data (link), B0034 is stronger than B0032. As a result, we didn’t obtain any visible bacterial colonies on agar plates. Our conslucion to this experiment is that additional regulation of MinC gene expression is very important to keep the system working.
