Team:UiOslo Norway/Achievements/Parts

From 2014.igem.org

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<h1>Parts</h1>
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<h3>Parts completed and sent to the iGEM headquarters</h3>
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<strong><a href ="http://parts.igem.org/Part:BBa_K1466001">BBa_K1466001</a> – Autotransporter signal peptide</strong><br>
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<p>Signal peptide guiding the autotransporter constructs to the SEC complex. The SEC complex is found on the inner membrane of the E.coli. The function of the SEC complex is to transport peptides/proteins from the intracellular space and over the inner membrane. The autotransporters are based in the outer membrane of the bacteria and thus needs to be transported over the inner membrane by the SEC complex before insertion into the outer membrane. The signal peptide will lead the unfolded amino acid sequence to the SEC complex.</p>
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<strong><a href ="http://parts.igem.org/Part:BBa_K1466000">BBa_K1466000</a> – LacZ Ω</strong><br>
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<p>Beta-galactosidase subunit with stop codon. This part will be express on the surface of the bacteria and bind to the alpha subunit expressed on the surface of a different bacteria. The subunit will be fused to the autotransporter Invfull and express with its C-term outwards from bacteria surface. We made this part based from the full length LacZ gene found in the kit (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K909006">BBa_K909006</a>).</p>
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<h3>Parts planned but not completed</h3>
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<strong>Beta-galactosidase Ω subunit without stop codon</strong><br>
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<p>This part will be expressed on the surface of the bacteria and bind to the alpha subunit expressed on the surface of a different bacteria. The subunit will be fused to the autotransporter Ehaj and express with its N-term outwards from bacteria surface. We planned to make this part based from the full length LacZ gene found in the kit (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K909006">BBa_K909006</a>).</p>
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<strong>EhaJ autotransporter</strong><br>
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<p>This autotransporter will express the N-term on the bacterial cell surface. We planned to use this to express the N-term of the galatosidase subunits on the cell surface. </p>
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<strong>Inverted autotransporter</strong><br>
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<p>This autotransporter will express its C-term on the bacteria cell surface. We planned to use this to express the C-term of our galactosidase subunits on the bacterial surface.</p>
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</li>
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<strong>Beta-galactosidase subunit without stop codon</strong><br>
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<p>This part will be expressed on the surface of the bacteria and bind to the alpha subunit expressed on the surface of a different bacteria. The subunit will be fused to the autotransporter Ehaj1 and express with its N-term outwards from bacteria surface. We planned to make this part based from the full length LacZ gene found in the kit (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K909006">BBa_K909006</a>).</p>
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<strong>N-term histidine tag</strong><br>
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<p>To isolate the subunits to show that they are binding and forming an actives enzyme in solution we were going to use Histidine tags. This part was not in the kit, but it has been made before. (<a href="http://parts.igem.org/Part:BBa_T2001">BBa_T2001</a>).</p>
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<img src="https://static.igem.org/mediawiki/2014/f/ff/Uioslo_norway_Uio.jpg" height="100px">
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<img src="https://static.igem.org/mediawiki/2014/8/87/Uioslo_norway_Nbs.jpg" height="100px">
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Latest revision as of 14:44, 17 October 2014

UiOslo IGEM 2014

Parts

Parts completed and sent to the iGEM headquarters

  • BBa_K1466001 – Autotransporter signal peptide

    Signal peptide guiding the autotransporter constructs to the SEC complex. The SEC complex is found on the inner membrane of the E.coli. The function of the SEC complex is to transport peptides/proteins from the intracellular space and over the inner membrane. The autotransporters are based in the outer membrane of the bacteria and thus needs to be transported over the inner membrane by the SEC complex before insertion into the outer membrane. The signal peptide will lead the unfolded amino acid sequence to the SEC complex.

  • BBa_K1466000 – LacZ Ω

    Beta-galactosidase subunit with stop codon. This part will be express on the surface of the bacteria and bind to the alpha subunit expressed on the surface of a different bacteria. The subunit will be fused to the autotransporter Invfull and express with its C-term outwards from bacteria surface. We made this part based from the full length LacZ gene found in the kit (BBa_K909006).

Parts planned but not completed

  • Beta-galactosidase Ω subunit without stop codon

    This part will be expressed on the surface of the bacteria and bind to the alpha subunit expressed on the surface of a different bacteria. The subunit will be fused to the autotransporter Ehaj and express with its N-term outwards from bacteria surface. We planned to make this part based from the full length LacZ gene found in the kit (BBa_K909006).

  • EhaJ autotransporter

    This autotransporter will express the N-term on the bacterial cell surface. We planned to use this to express the N-term of the galatosidase subunits on the cell surface.

  • Inverted autotransporter

    This autotransporter will express its C-term on the bacteria cell surface. We planned to use this to express the C-term of our galactosidase subunits on the bacterial surface.

  • Beta-galactosidase subunit without stop codon

    This part will be expressed on the surface of the bacteria and bind to the alpha subunit expressed on the surface of a different bacteria. The subunit will be fused to the autotransporter Ehaj1 and express with its N-term outwards from bacteria surface. We planned to make this part based from the full length LacZ gene found in the kit (BBa_K909006).

  • N-term histidine tag

    To isolate the subunits to show that they are binding and forming an actives enzyme in solution we were going to use Histidine tags. This part was not in the kit, but it has been made before. (BBa_T2001).

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