Difference between revisions of "Part:BBa K1022102"

 
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1022102 short</partinfo>
 
<partinfo>BBa_K1022102 short</partinfo>
Line 11: Line 10:
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1022102 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1022102 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 harbored 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 12:56, 3 October 2013

pT7: RBS: His6- SUMO: Signiferin

This part codes for the peptide 'Signiferin' 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 harbored 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].