Difference between revisions of "Part:BBa K418007"

 
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K418007 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K418007 SequenceAndFeatures</partinfo>
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<h2>Contribution</h2>
 +
<p>
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The CHS3 is a transmembrane protein originating from eukaryotes, which is challenging to integrate into prokaryotes, although the Northwestern team used the cDNA. Furthermore, the enzyme is post-translationally glycosylated, which makes it nearly impossible to express a functional enzyme [1].
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The sequence of the CHS3 further contains forbidden PstI and XbaI sites (typical standard RFC[10]) inside of the gene (XbaI at 1117 and 1674 and PstI at 2318), so that it is impractical for usage as BioBrick part. <br>
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To provide a functional Chitin synthase that is working in <i>E.coli</i>, the 2017 TU Darmstadt Team brought up Part [https://parts.igem.org/Part:BBa_K2380000 BBa_K2380000] encoding
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bacterial chitin synthase NodC to the registry.
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<br>
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<b>Group:</b> iGEM17_TU_Darmstadt
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<br>
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<b>Authors:</b> Lisa-Marie Brenner, Isabelle Feinauer, Werner Kleindienst, Cristina Kurzknabe, Isabelle Marquardt, Patrick Müller, Sven Storch
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<br>
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<b>Link to our part:</b> [https://parts.igem.org/Part:BBa_K2380000 BBa_K2380000]
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</p>
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<p>[1] Cos, T., Ford, R. A., Trilla, J. A., Duran, A., Cabib, E., and Roncero, C. (1998) Molecular analysis of Chs3p participation in chitin synthase III activity. The FEBS Journal, 256, 419 – 426
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DOI: 10.1046/j.1432-1327.1998.2560419.x
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</p>
  
  

Latest revision as of 16:28, 1 November 2017


chitin synthase (CHS3) gene of S. cerevisiae

Chitin synthase (CHS3) plays an essential role in chitin synthesis in S. cerevisiae.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1117
    Illegal XbaI site found at 1674
    Illegal PstI site found at 2318
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2318
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1158
    Illegal BglII site found at 1671
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1117
    Illegal XbaI site found at 1674
    Illegal PstI site found at 2318
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1117
    Illegal XbaI site found at 1674
    Illegal PstI site found at 2318
    Illegal AgeI site found at 229
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2590
    Illegal SapI site found at 980

Contribution

The CHS3 is a transmembrane protein originating from eukaryotes, which is challenging to integrate into prokaryotes, although the Northwestern team used the cDNA. Furthermore, the enzyme is post-translationally glycosylated, which makes it nearly impossible to express a functional enzyme [1]. The sequence of the CHS3 further contains forbidden PstI and XbaI sites (typical standard RFC[10]) inside of the gene (XbaI at 1117 and 1674 and PstI at 2318), so that it is impractical for usage as BioBrick part.
To provide a functional Chitin synthase that is working in E.coli, the 2017 TU Darmstadt Team brought up Part BBa_K2380000 encoding bacterial chitin synthase NodC to the registry.
Group: iGEM17_TU_Darmstadt
Authors: Lisa-Marie Brenner, Isabelle Feinauer, Werner Kleindienst, Cristina Kurzknabe, Isabelle Marquardt, Patrick Müller, Sven Storch
Link to our part: BBa_K2380000

[1] Cos, T., Ford, R. A., Trilla, J. A., Duran, A., Cabib, E., and Roncero, C. (1998) Molecular analysis of Chs3p participation in chitin synthase III activity. The FEBS Journal, 256, 419 – 426 DOI: 10.1046/j.1432-1327.1998.2560419.x