Difference between revisions of "Part:BBa K5237015"
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<partinfo>BBa_K5237327 short</partinfo> | <partinfo>BBa_K5237327 short</partinfo> | ||
− | This part can be used for | + | This part contains the N-terminus of intimin harbouring the anti-EGFR (wild-type 7D12) adhesin and can be used for surface display of anti-EGFR nanobodies on <i>E.coli</i>. |
+ | Salema et al., (2013) showed efficient presentation of nanobodies on the surface of <i>E.coli</i> K-12 cells by fusing them to the β domain of intimin. The β domain of intimin comes from the pNeae2 plasmid (addgene #168300) and encodes an N-terminal signal peptide for Sec-dependent translocation into the periplasm, a LysM domain that interacts with the peptidoglycan and provides anchoring, and a β-barrel that inserts into the outer membrane. | ||
+ | The coding sequence of the anti-EGFR nanobody is located in the C-terminus (that is exposed to the extracellular milieu) between the E tag (GAPVPYPDPLEP) and the myc tag (EQKLISEED). These tags can be utilized for purification or detection of the protein. | ||
+ | |||
<html> | <html> | ||
<div class="thumb"> | <div class="thumb"> | ||
<div class="thumbinner" style="width:50%;"> | <div class="thumbinner" style="width:50%;"> | ||
− | <img alt="FRET_tetR-Oct1" src="https://static.igem.wiki/teams/5237/wetlab-results/ | + | <img alt="FRET_tetR-Oct1" src="https://static.igem.wiki/teams/5237/wetlab-results/anti-myc-wb-intimin-myc.png" style="width:99%;" class="thumbimage"> |
<div class="thumbcaption"> | <div class="thumbcaption"> | ||
<i><b>Figure 3: Fluorescence measurement of mNeonGreen, mScarlet-I and FRET.</b> Fluorescence intensity of mNeonGreen | <i><b>Figure 3: Fluorescence measurement of mNeonGreen, mScarlet-I and FRET.</b> Fluorescence intensity of mNeonGreen |
Revision as of 08:52, 30 September 2024
No part name specified with partinfo tag.
This part contains the N-terminus of intimin harbouring the anti-EGFR (wild-type 7D12) adhesin and can be used for surface display of anti-EGFR nanobodies on E.coli. Salema et al., (2013) showed efficient presentation of nanobodies on the surface of E.coli K-12 cells by fusing them to the β domain of intimin. The β domain of intimin comes from the pNeae2 plasmid (addgene #168300) and encodes an N-terminal signal peptide for Sec-dependent translocation into the periplasm, a LysM domain that interacts with the peptidoglycan and provides anchoring, and a β-barrel that inserts into the outer membrane. The coding sequence of the anti-EGFR nanobody is located in the C-terminus (that is exposed to the extracellular milieu) between the E tag (GAPVPYPDPLEP) and the myc tag (EQKLISEED). These tags can be utilized for purification or detection of the protein.
Sequence and Features
No part name specified with partinfo tag.