Team:Berlin/Project

From 2014.igem.org

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               <div class="sub-content-project">
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                 <h2 class="sub-content-project-headline">week two!</h2>
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                 <h2 class="sub-content-project-headline">Week 4: 21.04.2014 – 27. 04.2014 </h2>
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                  Furthermore, we will work with other metal binding proteins such as metallothioneins and phytochelatin synthases in order to achieve nanoparticle synthesis. Once we have discovered the best way to magnetize E. coli bacteria, we will build and characterize suitable BioBricks that can be used by any research lab or iGEM team in the world in order to remote control the cellular movement.
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                    Minirep
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              </div>
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              <div class="sub-content-project">
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                <h2 class="sub-content-project-headline">Week 5: 28.04.2014 – 04.05.2014</h2>
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                    PCR Plasmid/Primar and Parameter check with Q5 Polymerase and Phusion Polymerase<br/>
 +
                    <br/>
 +
                    Miniprep of [pKD46+ DH5α] and [pKD4 + DH5 α]<br/>
 +
                    Genomic DNA extraction from Pseudomonas putida<br/>
 +
                    Enzyme digestion of different plasmids<br/>
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              </div>           
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              <div class="sub-content-project">
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                <h2 class="sub-content-project-headline">Week 6: 05.05.2014 – 11.05.2014</h2>
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                    Preculture of strains from Budisa strain databse in LB medium and Midiprep<br/>
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                    <br/>
 +
                    Canamycin cassette PCR of pKD4<br/>
 +
                    <br/>
 +
                    Restriction digest of Plasmid pKD4 and pKD46v<br/>
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              </div>           
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              <div class="sub-content-project">
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                <h2 class="sub-content-project-headline">Week 7: 12.05.2014 – 18.05.2014</h2>
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                    Biotransformation of Ferritin and pKD46 in Nissle and DH5 α strains
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              </div>           
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              <div class="sub-content-project">
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                <h2 class="sub-content-project-headline">Week 8: 19.05.2014 – 25.05.2014</h2>
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                    Gel extraction of FieF PCR<br/>
 +
                    <br/>
 +
                    Gene Knockout of FieF in [RV + pKD46] and [WM10+pKD46]<br/>
 +
                    <br/>
 +
                    Transformation of pKD46 in Nissle and MG 1655<br/>
 +
                    <br/>
 +
                    PCR of ATPCS and PPMT<br/>
 +
                    <br/>
 +
                    Digestion of pQE_80L for cloning<br/>
 +
              </div>
 +
              <div class="sub-content-project">
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                <h2 class="sub-content-project-headline">Week 9: 26.05.2014 – 01.06.2014</h2>
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                    Production of LB plates<br/>
 +
                    <br/>
 +
                    AMB-1 and Microfluidic Chip were picked up from Max-Plank Institute<br/>
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              </div>
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              <div class="sub-content-project">
 +
                <h2 class="sub-content-project-headline">Week 10: 02.06.2014 – 08.06.2014</h2>
 +
                    Colony PCR and analytical gel electrophoresis for identifying the right clones<br/>
 +
                    <br/>
 +
                    PCR of ATPCS and PPMT to identify the correct amplification parameter.<br/>
 +
                    <br/>
 +
                    Colony PCR for Knockout.<br/>
 +
                    <br/>
 +
                    PCR of ATPCS and PPMT with Q5 High Fidelity Mastermix.<br/>
               </div>
               </div>
               <div class="sub-content-project">
               <div class="sub-content-project">

Revision as of 21:34, 14 October 2014

1

What is it all about?

  As previous iGEM teams have shown, synthesizing fully functional magnetosomes in E. coli is highly difficult as more than 60 highly regulated genes are involved. As a more feasible alternative, we simply want to synthesize magnetic nanoparticles in E. coli in order to attract cells with strong magnetic fields.
Therefore we want to use different strategies including manipulation of the iron homeostasis of E. coli, expression of different metal binding proteins such as ferritins and metallothioneins as well as a high-throughput growth medium optimization.

Furthermore, we will work with other metal binding proteins such as metallothioneins and phytochelatin synthases in order to achieve nanoparticle synthesis. Once we have discovered the best way to magnetize E. coli bacteria, we will build and characterize suitable BioBricks that can be used by any research lab or iGEM team in the world in order to remote control the cellular movement.

2

Visualisation

  Animation, jo!

3

Detailes Description

  MORE DETAILS, jo!

4

Flow Chart

  Flowchart, jo!

5

The Results

  Results, jo!

6

Lab Summary

 

Week 1: 02.04.2014 – 06.04.2014

Cultivation of E. coli Nissle 1917

Genomic DNA extraction of E.coli Nissle strain

Production of BfR, FTNA1 and FTNA2
Restriction digest
PCR Purification
Ligation Assay
Transformation into DH5α-Cells

Week 2: 07.04.2014 – 13.04.2014

Colony PCR to check the results

Bfr, FTNA 1, FTNA 2 primar designed for amplification

Transformation of pQE_80L into DH5α

Cultivation of BfR, FTNA 1, FTNA 2 in LB

Week 3: 14.04.2014 – 20.04.2014

Miniprep of the cells from Week 2

DNA concentration determination and sequencing

Expression of BfR, FTNA 1, FTNA 2 and induction with IPTG

Production of a Mutaflor-Supression culture and streaking

Week 4: 21.04.2014 – 27. 04.2014

Minirep

Week 5: 28.04.2014 – 04.05.2014

PCR Plasmid/Primar and Parameter check with Q5 Polymerase and Phusion Polymerase

Miniprep of [pKD46+ DH5α] and [pKD4 + DH5 α]
Genomic DNA extraction from Pseudomonas putida
Enzyme digestion of different plasmids

Week 6: 05.05.2014 – 11.05.2014

Preculture of strains from Budisa strain databse in LB medium and Midiprep

Canamycin cassette PCR of pKD4

Restriction digest of Plasmid pKD4 and pKD46v

Week 7: 12.05.2014 – 18.05.2014

Biotransformation of Ferritin and pKD46 in Nissle and DH5 α strains

Week 8: 19.05.2014 – 25.05.2014

Gel extraction of FieF PCR

Gene Knockout of FieF in [RV + pKD46] and [WM10+pKD46]

Transformation of pKD46 in Nissle and MG 1655

PCR of ATPCS and PPMT

Digestion of pQE_80L for cloning

Week 9: 26.05.2014 – 01.06.2014

Production of LB plates

AMB-1 and Microfluidic Chip were picked up from Max-Plank Institute

Week 10: 02.06.2014 – 08.06.2014

Colony PCR and analytical gel electrophoresis for identifying the right clones

PCR of ATPCS and PPMT to identify the correct amplification parameter.

Colony PCR for Knockout.

PCR of ATPCS and PPMT with Q5 High Fidelity Mastermix.

week one!

Furthermore, we will work with other metal binding proteins such as metallothioneins and phytochelatin synthases in order to achieve nanoparticle synthesis. Once we have discovered the best way to magnetize E. coli bacteria, we will build and characterize suitable BioBricks that can be used by any research lab or iGEM team in the world in order to remote control the cellular movement.
7

Notebook

  Notebook, jo!