Difference between revisions of "Part:BBa K1022101"
Line 1: | Line 1: | ||
− | |||
__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1022101 short</partinfo> | <partinfo>BBa_K1022101 short</partinfo> | ||
Line 11: | Line 10: | ||
<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K1022101 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1022101 SequenceAndFeatures</partinfo> | ||
+ | |||
+ | ===Characterization=== | ||
+ | '''For more info, visit [http://2013.igem.org/Team:TU-Delft/Peptides#SUMO TU Delft iGEM13 Wiki]''' | ||
+ | |||
+ | '''Peptide production : ''' | ||
+ | |||
+ | In the project by iGEM 13 Team TU Delft, a SUMO-peptide fusion was opted as a suitable expression system as they make the fusion proteins more soluble. The peptide by itself is not soluble in the cytoplasm but making a fusion of peptide with Small Ubiquitin like Modifiers (SUMO) will increase the solubility of the peptide, thus increasing the cytoplasmic fraction of the peptide. | ||
+ | |||
+ | A gene was constructed in such a way that the SUMO-peptide production was driven by the strong T7 phage promoter. This gene containing plasmid was harboured in a BL21(DE3) strain that has lac promoter driven T7 polymerase. Upon induction by IPTG the SUMO peptide fusion is produced as a soluble protein fraction. | ||
+ | |||
+ | The protocol can be seen [http://2013.igem.org/Team:TU-Delft/Protocol_10#protocol_10 here]. | ||
Revision as of 12:57, 3 October 2013
pT7: RBS: His6 - SUMO: MaximinH5
This part codes for the peptide 'Maximin H5' tagged with 'His-6-SUMO' molecule. Its production is triggered by IPTG (through pT7 promoter).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 46
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 170
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 355
- 1000COMPATIBLE WITH RFC[1000]
Characterization
For more info, visit [http://2013.igem.org/Team:TU-Delft/Peptides#SUMO TU Delft iGEM13 Wiki]
Peptide production :
In the project by iGEM 13 Team TU Delft, a SUMO-peptide fusion was opted as a suitable expression system as they make the fusion proteins more soluble. The peptide by itself is not soluble in the cytoplasm but making a fusion of peptide with Small Ubiquitin like Modifiers (SUMO) will increase the solubility of the peptide, thus increasing the cytoplasmic fraction of the peptide.
A gene was constructed in such a way that the SUMO-peptide production was driven by the strong T7 phage promoter. This gene containing plasmid was harboured in a BL21(DE3) strain that has lac promoter driven T7 polymerase. Upon induction by IPTG the SUMO peptide fusion is produced as a soluble protein fraction.
The protocol can be seen [http://2013.igem.org/Team:TU-Delft/Protocol_10#protocol_10 here].