Difference between revisions of "Part:BBa K3117006"
Line 2: Line 2:
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K3117006 short</partinfo>
 
<partinfo>BBa_K3117006 short</partinfo>
−
 
−
The Igk leader is a signaling sequence, directing the protein into the secretory pathway (Feng, Gong, & Dimitrov, 2011).
 
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
 +
The Igk leader is a signaling sequence, directing the protein into the secretory pathway (Feng et al., 2011). By adding this signaling peptide the protein of interest can be secreted in the supernatant after transfection and the basis for a proper protein folding can provided. Prior to secretion the Igk leader is cleaved of.
 +
 +
===Characterization===
 +
In our project the Igk leader was added to three proteins (<partinfo>BBa_K3117026</partinfo>, <partinfo>BBa_K3117030</partinfo>, <partinfo>BBa_K3117029</partinfo>) for the secretion of our desired proteins into the supernatant, after transfection into HEK 293T cells.
  
 +
[[File:T--FAU_Erlangen--Bild_Sequencing_Ernte_Results_Composite_part.png|thumb|center|200px|'''Figure 2''': Western blot of the harvest after transfection of HEK 293T cells. Anti His-antibody was used for detection]]
  
 +
In Fig. 1 the western blot of the harvest of HEK 293T cells can be seen. Those proteins were detected via their His-Tag. The presence of the proteins in the supernatant shows, that the Igk leader was able to direct all three proteins into the secretory way.
  
 +
<partinfo></partinfo>
 
<!-- -->
 
<!-- -->
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 23:18, 21 October 2019


Igk leader sequence

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

By adding this signaling peptide the protein of interest can be secreted in the supernatant after transfection and the basis for a proper protein folding can provided. Prior to secretion the Igk leader is cleaved of. Igk leader is normally fused to the N-terminus of the protein of interest. The Team 2019 FAU_Erlangen used the Igk leader sequence to secrete their proteins (bispecific antibodies)after HEK293T cell transfection.