Team:Heidelberg/Safety

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                                 <h1><span style="font-size:180%;color:#9e90a3;">Safety.</span><span class="text-muted" style="font-size:100%"> We take good care of you!</span></h1>
                                 <h1><span style="font-size:180%;color:#9e90a3;">Safety.</span><span class="text-muted" style="font-size:100%"> We take good care of you!</span></h1>
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                                 <p style="text-align:justify">Since the Daci-Cluster genes of <i>D. acidovorans</i>, which are considered to be crucial for the production of delftibactin consist of more than 58 kb, we had to think of a method capable of trasferring this great amount of genetic information to our host organism <i>E. coli</i>. Being confronted with this task, we considered the usage of so called <i>"Bacterial Artificial Chromosomes (BACs)"</i> or <i>"Cosmids</i>. But finally, we decided to face the challege by splitting up the crucial genes of the cluster into convetional plasmids. One containing the gigantic gene "DelH", which comprises 18 kb and another containing all residual parts, herein referred to as "DelRest". Additionally, we planed to integrate <i>D. acidovorans</i>'s methyl-malonyl-CoA-pathway, which is a crucial co-factor for one of the PKS domains and hence, for succesful delftibactin production. Because we were unfortunately not able to genomically integrate the pathway after several attempts, we decided to create a third plasmid not only containing the pathway, but as well a permeability device (BBa_...) for enabeling our bacteria to secrete the desired product. These three plasmids were constructed in different subgroups and therefore have separated labjournals. Follow the links below to read more!
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                                 <p style="text-align:justify">The uttermost anxiety of those who do not work with genetically modified organisms is concerning safety. We face this critical issue with a tripartite approach. Safety precautions concerning our project can be broken down to the following three topics: Firstly, safety considerations about the wet-lab project, secondly, the general lab safety and thirdly, software safety.
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In all of the three strands, we deliberated useful approaches and searched communication with safety advisors. Herewith, we wanted to get additional input which we could use for building up our opinion on which parts of the project could be critical or which measures would have to be taken.
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Revision as of 10:18, 4 October 2013


Safety. We take good care of you!

The uttermost anxiety of those who do not work with genetically modified organisms is concerning safety. We face this critical issue with a tripartite approach. Safety precautions concerning our project can be broken down to the following three topics: Firstly, safety considerations about the wet-lab project, secondly, the general lab safety and thirdly, software safety. In all of the three strands, we deliberated useful approaches and searched communication with safety advisors. Herewith, we wanted to get additional input which we could use for building up our opinion on which parts of the project could be critical or which measures would have to be taken.

Del H

The largest part of the pathway was amplified in subdivided fragments and subsequently assembled to a 22 kb plasmid, which was later co-transformed with the other plasmids.

Visit Labjournal

Del Rest

"DelRest" consists of the genes DelA, B, C, D, E, F, G, L, O ad P, which encode NRPS- as well as PKS (polyketide synthethase) modules. We divided these genes on the cluster into certain fragments, which were amplified with Gibson-overlaps for subsequent assembly.

Visit Labjournal

Methylmalonyl-CoA Pathway

After our attempts on trying to genomically integrate this pathway into E. coli didn't work out well, we created a third plasmid, on which we wanted to integrate it. Additionally we therafter built in a permeability device from the parts-registry for delftibactin secretion into the medium.

Visit Labjournal

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