Difference between revisions of "Part:BBa K1022101"
Line 21: Line 21:
  
 
The protocol can be seen  [http://2013.igem.org/Team:TU-Delft/Protocol_10#protocol_10 here].  
 
The protocol can be seen  [http://2013.igem.org/Team:TU-Delft/Protocol_10#protocol_10 here].  
 +
 +
'''MIC determination:'''
 +
 +
The MIC of peptide on ''S. delphini, B. subtilis and E. coli'' was also done. This characterization can be seen in construct [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1022110  BBa_K1022110].
 +
 +
Also visit the [http://2013.igem.org/Team:TU-Delft/PeptideCharacterization Peptide characterization] page on TU Delft iGEM13 Wiki to know more about the MIC experiments.
  
  

Revision as of 13:13, 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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 46
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 170
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 355
  • 1000
    COMPATIBLE 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].

MIC determination:

The MIC of peptide on S. delphini, B. subtilis and E. coli was also done. This characterization can be seen in construct BBa_K1022110.

Also visit the [http://2013.igem.org/Team:TU-Delft/PeptideCharacterization Peptide characterization] page on TU Delft iGEM13 Wiki to know more about the MIC experiments.