Difference between revisions of "Part:BBa K2848001"
Line 5: Line 5:
 
This Part allows for the production of chitosan in a single cell with the need of only one plasmid to be transformed. We combined two parts from the iGEM Team Darmstadt 2017 (ChiTUCare), NodC (chitin synthase, BBa_2380002) and COD (chitin deacetylase, BBa_2380043). The COD was improved by adding a His-Tag for easy purification after expression in E.coli BL21 DE3. The use of the BL21 DE3 strain is important, since the COD-ORF is under the control of an IPTG inducible T7 promotor.
 
This Part allows for the production of chitosan in a single cell with the need of only one plasmid to be transformed. We combined two parts from the iGEM Team Darmstadt 2017 (ChiTUCare), NodC (chitin synthase, BBa_2380002) and COD (chitin deacetylase, BBa_2380043). The COD was improved by adding a His-Tag for easy purification after expression in E.coli BL21 DE3. The use of the BL21 DE3 strain is important, since the COD-ORF is under the control of an IPTG inducible T7 promotor.
  
−
File:T--Stuttgart--Bba_K2848001_Map.png|200px|thumb|left|Plasmid map was created with Snapgene.
+
<img align="center" style='height: 200px; width: 200px; object-fit: contain' src="https://static.igem.org/mediawiki/2018/b/bb/T--Stuttgart--Bba_K2848001_Map.png" alt="Plasmid map was created with Snapgene.">
  
 
By using two different induction systems, Arabinose for NodC and IPTG for COD, a sequential expression of the emzymes is possible. This might optimize the production of chitosan wich has a very low solubility in aqueos solutions.
 
By using two different induction systems, Arabinose for NodC and IPTG for COD, a sequential expression of the emzymes is possible. This might optimize the production of chitosan wich has a very low solubility in aqueos solutions.

Revision as of 14:17, 9 October 2018


NodC+COD-HisTag

This Part allows for the production of chitosan in a single cell with the need of only one plasmid to be transformed. We combined two parts from the iGEM Team Darmstadt 2017 (ChiTUCare), NodC (chitin synthase, BBa_2380002) and COD (chitin deacetylase, BBa_2380043). The COD was improved by adding a His-Tag for easy purification after expression in E.coli BL21 DE3. The use of the BL21 DE3 strain is important, since the COD-ORF is under the control of an IPTG inducible T7 promotor.

<img align="center" style='height: 200px; width: 200px; object-fit: contain' src="T--Stuttgart--Bba_K2848001_Map.png" alt="Plasmid map was created with Snapgene.">

By using two different induction systems, Arabinose for NodC and IPTG for COD, a sequential expression of the emzymes is possible. This might optimize the production of chitosan wich has a very low solubility in aqueos solutions.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1613
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961