Part:BBa_K418007

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
Author: Thomas Georg Wagner
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