Difference between revisions of "Part:BBa K2848001"
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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. | ||
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− | + | <img style='height: 60%; width: 60%; object-fit: contain' src="https://static.igem.org/mediawiki/2018/4/40/T--Stuttgart--BBa_K2848001_Map.png" alt="Plasmid map was created with Snapgene."> | |
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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. | ||
Latest revision as of 14:31, 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.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
Illegal AgeI site found at 1613 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961