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During the mentorship we picked certain topics for every skype meeting. Those topics covered technical and practical advices concerning the wiki and important points for human practice among others. Together we discussed all the gold medal criteria and gave tips how to fulfill them. Furthermore we discussed organizational aspects of travel and stay in Boston. During one skype call our HTML programming expert Maximilian Edich answered questions concerning HTML coding. <br> | During the mentorship we picked certain topics for every skype meeting. Those topics covered technical and practical advices concerning the wiki and important points for human practice among others. Together we discussed all the gold medal criteria and gave tips how to fulfill them. Furthermore we discussed organizational aspects of travel and stay in Boston. During one skype call our HTML programming expert Maximilian Edich answered questions concerning HTML coding. <br> | ||
− | In return for the mentorship, iGEM UNIFI helped us characterizing two BioBricks. To make sure that <i>Escherichia coli</i> is able to take up the unnatural nucleoside triphosphates from the cultivation media we had to introduce a heterologous transporter. This is due to a lack of nucleotide transporters in <i>E. coli</i>. One of the BioBricks encodes a complete nucleotide transporter | + | In return for the mentorship, iGEM UNIFI helped us characterizing two BioBricks. To make sure that <i>Escherichia coli</i> is able to take up the unnatural nucleoside triphosphates from the cultivation media we had to introduce a heterologous transporter. This is due to a lack of nucleotide transporters in <i>E. coli</i>. One of the BioBricks encodes a complete nucleotide transporter <i>pt</i>NTT2 (BBa_K2201000) originated from the algae <i>Phaeodactylum tricornutum</i>. The second BioBrick is a truncated version missing the N-terminal signal peptide (BBa_K2201001). This N-terminal signal peptide leads to some kind of toxicity in <i>E. coli</i>. Through cultivation experiments we wanted to investigate the extent of the toxicity by comparing the growth of the strain expressing the full version of <i>Pt</i>NTT2 to the ones expressing the truncated version. <br> |
We started to cultivate the different strains in 50 mL media using flasks and measured the OD<SUB>600</SUB> every 30 minutes during the exponential growing phase. Due to manual measurements our results showed big error values for the maximum growing rate µmax. This makes it hard to get a valid conclusion. iGEM UNIFI has the capacity to do the same cultivation experiment using a microscale bioreactor. This ensures automatic measurements for OD<SUB>600</SUB> values which would decrease errors concerning µmax. This characterization from iGEM UNIFI would lead to a more accurate estimation of the toxicity of a full length version compared to a truncated version of <i>Pt</i>NTT2. | We started to cultivate the different strains in 50 mL media using flasks and measured the OD<SUB>600</SUB> every 30 minutes during the exponential growing phase. Due to manual measurements our results showed big error values for the maximum growing rate µmax. This makes it hard to get a valid conclusion. iGEM UNIFI has the capacity to do the same cultivation experiment using a microscale bioreactor. This ensures automatic measurements for OD<SUB>600</SUB> values which would decrease errors concerning µmax. This characterization from iGEM UNIFI would lead to a more accurate estimation of the toxicity of a full length version compared to a truncated version of <i>Pt</i>NTT2. | ||
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<h3> Collaboration - Lokalization study and part exchange with <a href="https://2017.igem.org/Team:CU-Boulder">CU Boulder 2017</a> </h3> | <h3> Collaboration - Lokalization study and part exchange with <a href="https://2017.igem.org/Team:CU-Boulder">CU Boulder 2017</a> </h3> | ||
<article> | <article> | ||
− | This year the iGEM team CU Boulder works with artificial compartments which consist of shell proteins containing a photoswitch amino acid. These compartments are supposed to break up into separate shell proteins releasing the protein captured inside the compartment. This could potentially act as next-generation drug delivery systems, biosensors, or as a solution to sequester diffuse and harmful environmental toxins. The iGEM team CU Boulder 2016 submitted the BioBrick containing the aminoacyl tRNA synthetase (aaRS) for the photoswitch amino acid. This year’s team helped us to realize our photoswitching project by sending us their aaRS and a small amount of the amino acid AzeF. In return, we transformed their parts EutS (BBa_K2129001) and EutC-tagged FusionRed (BBa_XX) in E.coli DH5α and performed localization studies by fluorescence microscopy with our confocal laser scan microscope. | + | This year the iGEM team CU Boulder works with artificial compartments which consist of shell proteins containing a photoswitch amino acid. These compartments are supposed to break up into separate shell proteins releasing the protein captured inside the compartment. This could potentially act as next-generation drug delivery systems, biosensors, or as a solution to sequester diffuse and harmful environmental toxins. The iGEM team CU Boulder 2016 submitted the BioBrick containing the aminoacyl tRNA synthetase (aaRS) for the photoswitch amino acid. This year’s team helped us to realize our photoswitching project by sending us their aaRS and a small amount of the amino acid AzeF. In return, we transformed their parts EutS (BBa_K2129001) and EutC-tagged FusionRed (BBa_XX) in <i>E.coli</i> DH5α and performed localization studies by fluorescence microscopy with our confocal laser scan microscope. |
</article> | </article> | ||
<div class="figure medium"> | <div class="figure medium"> | ||
<img class="figure image" src="https://static.igem.org/mediawiki/2017/9/92/T--Bielefeld-CeBiTec--YKE_FusRed.gif"> | <img class="figure image" src="https://static.igem.org/mediawiki/2017/9/92/T--Bielefeld-CeBiTec--YKE_FusRed.gif"> | ||
− | <p class="figure subtitle"><b>Figure 2:</b> 3D-Animation of the fluorescence signal of E.coli cells transformed with EutC-tagged FusionRed from CU Boulder. The fluorescence is present in the whole cell.</p> | + | <p class="figure subtitle"><b>Figure 2:</b> 3D-Animation of the fluorescence signal of <i>E.coli</i> cells transformed with EutC-tagged FusionRed from CU Boulder. The fluorescence is present in the whole cell.</p> |
</div> | </div> | ||
<div class="figure medium"> | <div class="figure medium"> | ||
<img class="figure image" src="https://static.igem.org/mediawiki/2017/4/4d/T--Bielefeld-CeBiTec--YKE_EutS_Co.gif"> | <img class="figure image" src="https://static.igem.org/mediawiki/2017/4/4d/T--Bielefeld-CeBiTec--YKE_EutS_Co.gif"> | ||
− | <p class="figure subtitle"><b>Figure 3:</b> 3D-Animation of the fluorescence signal of three E.coli cells cotransformed with shell protein EutS and EutC-tagged FusionRed from CU Boulder. The fluorescence is concentrated in the EutS-compartments.</p> | + | <p class="figure subtitle"><b>Figure 3:</b> 3D-Animation of the fluorescence signal of three <i>E.coli</i> cells cotransformed with shell protein EutS and EutC-tagged FusionRed from CU Boulder. The fluorescence is concentrated in the EutS-compartments.</p> |
</div><div class="bevel bl"></div> | </div><div class="bevel bl"></div> | ||
<article> | <article> |
Revision as of 20:16, 28 October 2017
Overview
Collaboration – Mentoring iGEM team UNIFI 2017
Figure 1: Skype meetings with iGEM UNIFI for a two-way collaboration.
In return for the mentorship, iGEM UNIFI helped us characterizing two BioBricks. To make sure that Escherichia coli is able to take up the unnatural nucleoside triphosphates from the cultivation media we had to introduce a heterologous transporter. This is due to a lack of nucleotide transporters in E. coli. One of the BioBricks encodes a complete nucleotide transporter ptNTT2 (BBa_K2201000) originated from the algae Phaeodactylum tricornutum. The second BioBrick is a truncated version missing the N-terminal signal peptide (BBa_K2201001). This N-terminal signal peptide leads to some kind of toxicity in E. coli. Through cultivation experiments we wanted to investigate the extent of the toxicity by comparing the growth of the strain expressing the full version of PtNTT2 to the ones expressing the truncated version.
We started to cultivate the different strains in 50 mL media using flasks and measured the OD600 every 30 minutes during the exponential growing phase. Due to manual measurements our results showed big error values for the maximum growing rate µmax. This makes it hard to get a valid conclusion. iGEM UNIFI has the capacity to do the same cultivation experiment using a microscale bioreactor. This ensures automatic measurements for OD600 values which would decrease errors concerning µmax. This characterization from iGEM UNIFI would lead to a more accurate estimation of the toxicity of a full length version compared to a truncated version of PtNTT2.
Collaboration - Lokalization study and part exchange with CU Boulder 2017
Figure 2: 3D-Animation of the fluorescence signal of E.coli cells transformed with EutC-tagged FusionRed from CU Boulder. The fluorescence is present in the whole cell.
Figure 3: 3D-Animation of the fluorescence signal of three E.coli cells cotransformed with shell protein EutS and EutC-tagged FusionRed from CU Boulder. The fluorescence is concentrated in the EutS-compartments.
We are very happy that they provided their aaRS to us to expand our toolkit and we hope that our results of the localization study are helpful for their further work.
Networking
22.04 - March for Science in Bonn
Figure 4: Daniel Bergen at the March for Science in Bonn at the 22nd April with members of the team Cologne Duesseldorf and other iGEM teams.
13.05, 15.07 - Meetings with team Cologne Duesseldorf
Figure 5: Impressions of our meet ups with the team Cologne Duesseldorf. Our visit in Dusseldorf left and the meet up in Bielefeld right.
30.06 - German Meet-up in Dresden
Figure 6: Our speaker Chris Whitford at the German meet-up in Dresden, presenting our project to the public for the first time.
06-08.07 - European Meet-up in Delft
Figure 7: Six of our team members at the European meet-up in delft.
Postcards
Figure 8: Collection of the postcards of iGEM teams from around the world. Our card is the black one in the lower middle.