Team:Penn/SurfaceDisplayBBa
From 2012.igem.org
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- | We sought to create a system in which iGEM teams and labs can display any protein of their choosing on the surface of E. coli. We a novel Suface Display BioBrick (BBa K811004) using the N and C terminal domains of the Ice Nucleation Protein (INPNC) which allows iGEM teams to fuse any desired protein to INPNC using a simple ligation protocol with BamHI and PstI restriction sites.</p> | + | We sought to create a system in which iGEM teams and labs can display any protein of their choosing on the surface of E. coli. We engineered a novel Suface Display BioBrick (BBa K811004) using the N and C terminal domains of the Ice Nucleation Protein (INPNC) which allows iGEM teams to fuse any desired protein to INPNC using a simple ligation protocol with BamHI and PstI restriction sites.</p> |
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Revision as of 13:53, 26 October 2012
We sought to create a system in which iGEM teams and labs can display any protein of their choosing on the surface of E. coli. We engineered a novel Suface Display BioBrick (BBa K811004) using the N and C terminal domains of the Ice Nucleation Protein (INPNC) which allows iGEM teams to fuse any desired protein to INPNC using a simple ligation protocol with BamHI and PstI restriction sites.
As a preliminary proof-of-concept for our INPNC-enabled system, we displayed the red fluorescent protein mCherry on the outer membrane of E. coli BL21. After sonication and centrifugation of induced cells, almost all of the mCherry was localized in the membrane fraction when fused to INPNC, whereas in the control Intein-mCherry fusion (which exhibits cytoplasmic localization), all of the mCherry was contained in the lysate (Figure 1).
![](https://static.igem.org/mediawiki/2012/a/a2/MCherry-vs-INPNC-mCherry.jpg)
Figure 1