Revision as of 16:36, 14 September 2015

Conclusions

Our results for the different subprojects led us to the following conclusions:

Correct folding study of target protein

Structure analysis of our targets and their interactions

Investigation of P3 threshold for E. coli resistance

Some difficulties were found with the expression of P3 and the resulting washout of the phages. The plasmid showed itself stable during the whole cultivation which makes the next step a cultivation without antibiotics possible and highly desirable.

Further experiments regarding the plasmid stability should be performed, as well as reducing the yeast extract from the medium by identifying the missing element in the medium. A special expression analysis has to be done on P3 in relation to the IPTG-concentration.

In order to ensure proper antibiotic denaturation the cultivated medium had to be sterilized for 20 minutes at 121 °C. Further experiments are necessary to test the need of antibiotics for plasmid stability in the system. Since the antibiotic resistance of the recombinant E. coli strand is based on chloramphenicol degradation..

Additionally, the synthesized enzyme is secreted into the cultivation medium. This might lead to a complete loss of antibiotic function and therefore allow plasmid free E. coli to reproduce. As a result, the plasmid free cells might accumulate inside the CSR. Thus it is necessary to compare the plasmid stability of antibiotic free and antibiotic enriched media. Accounting to the previous reasons the influence of the chloramphenicol on plasmid stability might be negligible. As a result other ways to support plasmid stability or antibiotic free systems might be used for further experiments.

Conversion of BACTH into an iGEM standard and analysis of function

Set up of flow system

The continuous cultivation and CFP expression analysis gave a first idea of the steps needed to perform in order to obtain a continuous evolutionary process. It was possible to prove that the continuous flow system works as expected and that the phages are able to infect the E. coli.

All in all, the initial experiments helped us to grasp the idea of how such experiments can look like and where they are headed. We could show that our flow system works with the phages and the waste could be reduced to a minimum.

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    <h2 id="setup">Set up of flow system</h2>
    <a href="https://2015.igem.org/Team:TU_Dresden/Project/Conclusions#setup"></a>
-<p style="line-height:1.8">The continues cultivation and CFP expression analysis gave a first idea of the steps which needed to be performed in order of getting for a continuous evolutionary process. It was possible to proof that the continuousues flow system works as expected and that the phages are able to infect the <i>E. coli</i>. </p>
+<p style="line-height:1.8">The continuous cultivation and CFP expression analysis gave a first idea of the steps needed to perform in order to obtain a continuous evolutionary process. It was possible to prove that the continuous flow system works as expected and that the phages are able to infect the <i>E. coli</i>. </p>
-<p style="line-height:1.8">All in all the initial experiments helped us to get a good idea what the experiments can look like and where they are heading. We could show that our flow system works with the phages and the waste could be reduced to a minimum. </p>
+<p style="line-height:1.8">All in all, the initial experiments helped us to grasp the idea of how such experiments can look like and where they are headed. We could show that our flow system works with the phages and the waste could be reduced to a minimum. </p>
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