Difference between revisions of "Team:Bielefeld-CeBiTec/Basic Part"

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The broad range nucleotide transporter <i>Pt</i>NTT2 from <i>Phaeodactylum tricornutum</i> facilitates the counter exchange of nucleotide triphosphates (NTPs) and nucleoside diphosphates (NDPs), importing NTPs into the plastid. Expressed in <i>E. coli</i>, it allows the uptake of various nucleotides from the cultivation media. This function is essential for the development of a semisynthetic organism, which uses externally provided unnatural nucleotides for replication of its semisynthetic DNA. For our project, we characterized the structure, kinetics, dynamics and subcellular localization of <i>Pt</i>NTT2 and different <i>Pt</i>NTT2 variants. Therefore, we applied  bioinformatic prediction tools, confocal laser scanning microscopy, SDS-PAGE, Western Blot, MALDI-TOF as well as HPLC-MS. All <i>Pt</i>NTT2 variants were also cultivated to investigate the effect of <i>Pt</i>NTT2 expression on the growth rate. In the future, advanced endosymbiotic systems or novel biosafety mechanisms can be developed based on this part.  
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The broad range nucleotide transporter <i>Pt</i>NTT2 from <i>Phaeodactylum tricornutum</i> facilitates the counter exchange of nucleotide triphosphates (NTPs) and nucleoside diphosphates (NDPs), importing NTPs into the plastid. Expressed in <i>E. coli</i>, it allows the uptake of various nucleotides from the cultivation media. This function is essential for the development of a semisynthetic organism, which uses externally provided unnatural nucleotides for replication of its semisynthetic DNA. For our project, we characterized the structure, kinetics, dynamics and subcellular localization of <i>Pt</i>NTT2 and different <i>Pt</i>NTT2 variants. Therefore, we applied  bioinformatic prediction tools, confocal laser scanning microscopy, SDS-PAGE, Western Blot, MALDI-TOF as well as HPLC-MS. All <i>Pt</i>NTT2 variants were also cultivated to investigate the effect of <i>Pt</i>NTT2 expression on the growth rate. In the future, advanced endosymbiotic systems or novel biosafety mechanisms can be developed based on this part.  
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<td><a class="noul_link part_link" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2201004">BBa_K2201004</a></td>
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<td>Coding</td>
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<td>Nucleotide Transporter <i>Pt</i>NTT2 from <i>Phaeodactylum tricornutum</i></td>
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<td>Christopher M. Whitford</td>
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<td align="right">1728</td>
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<td><a class="noul_link part_link" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2201000">BBa_K2201000</a></td>
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<td>Coding</td>
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<td>Nucleotide Transporter <i>Pt</i>NTT2 with lacUV5 Promoter and RBS</td>
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<td>Christopher M. Whitford</td>
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<td align="right">1777</td>
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<td><a class="noul_link part_link" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2201001">BBa_K2201001</a></td>
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<td>Coding</td>
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<td>Truncated version <i>Pt</i>NTT2(66-575) with lacUV5 and RBS</td>
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<td>Christopher M. Whitford</td>
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<td align="right">1585</td>
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<td><a class="noul_link part_link" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2201002">BBa_K2201002</a></td>
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<td>Coding</td>
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<td><i>Pt</i>NTT2 tagged with GFP and a cMyc linker</td>
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<td>Christopher M. Whitford</td>
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<td align="right">2518</td>
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<td><a class="noul_link part_link" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2201003">BBa_K2201003</a></td>
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<td>Coding</td>
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<td><i>Pt</i>NTT2(66-575) tagged with GFP and a cMyc linker</td>
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<td>Christopher M. Whitford</td>
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<td align="right">2326</td>
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<td><a class="noul_link part_link" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2201005">BBa_K2201005</a></td>
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<td>Coding</td>
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<td>Truncated version PtNTT2(31-575) with lacUV5 and RBS</td>
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<td>Christopher M. Whitford</td>
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<td align="right">1690</td>
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Revision as of 07:12, 31 October 2017

Basic Parts
The broad range nucleotide transporter PtNTT2 from Phaeodactylum tricornutum facilitates the counter exchange of nucleotide triphosphates (NTPs) and nucleoside diphosphates (NDPs), importing NTPs into the plastid. Expressed in E. coli, it allows the uptake of various nucleotides from the cultivation media. This function is essential for the development of a semisynthetic organism, which uses externally provided unnatural nucleotides for replication of its semisynthetic DNA. For our project, we characterized the structure, kinetics, dynamics and subcellular localization of PtNTT2 and different PtNTT2 variants. Therefore, we applied bioinformatic prediction tools, confocal laser scanning microscopy, SDS-PAGE, Western Blot, MALDI-TOF as well as HPLC-MS. All PtNTT2 variants were also cultivated to investigate the effect of PtNTT2 expression on the growth rate. In the future, advanced endosymbiotic systems or novel biosafety mechanisms can be developed based on this part.
Name Type Description Designer Length
BBa_K2201004 Coding Nucleotide Transporter PtNTT2 from Phaeodactylum tricornutum Christopher M. Whitford 1728
BBa_K2201000 Coding Nucleotide Transporter PtNTT2 with lacUV5 Promoter and RBS Christopher M. Whitford 1777
BBa_K2201001 Coding Truncated version PtNTT2(66-575) with lacUV5 and RBS Christopher M. Whitford 1585
BBa_K2201002 Coding PtNTT2 tagged with GFP and a cMyc linker Christopher M. Whitford 2518
BBa_K2201003 Coding PtNTT2(66-575) tagged with GFP and a cMyc linker Christopher M. Whitford 2326
BBa_K2201005 Coding Truncated version PtNTT2(31-575) with lacUV5 and RBS Christopher M. Whitford 1690