Difference between revisions of "Part:BBa K5237015"
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<partinfo>BBa_K5237327 short</partinfo>
 
<partinfo>BBa_K5237327 short</partinfo>
  
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This part can be used for the display of nanobodies on the surface of <i>E.coli</i>. Salema, V. et al, (2013) showed efficient presentation of nanobodies on the surface of <i>E.coli</i> K-12 cells by fusing them to the &#946; domain of Intimin. The part 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 &#946;-barrel that inserts into the outer membrane. The C-terminus is exposed to the extracellular milieu and contains a myc tag (EQKLISEED) that can be used for purification or detection of full-length protein.  
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This part can be used for the display of nanobodies on the surface of <i>E.coli</i>. Salema, V. et al, (2013) showed efficient presentation of nanobodies on the surface of <i>E.coli</i> K-12 cells by fusing them to the &#946; domain of Intimin. The part 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 &#946;-barrel that inserts into the outer membrane. The C-terminus is exposed to the extracellular milieu and contains a myc tag (EQKLISEED) that can be used for purification or detection of full-length protein.
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<html>
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<div class="thumb">
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    <div class="thumbinner" style="width:50%;">
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      <img alt="FRET_tetR-Oct1" src="https://static.igem.wiki/teams/5237/wetlab-results/sist-fret-final.svg" style="width:99%;" class="thumbimage">
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      <div class="thumbcaption">
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        <i><b>Figure 3: Fluorescence measurement of mNeonGreen, mScarlet-I and FRET.</b> Fluorescence intensity of mNeonGreen
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          (ex. 490 nm, em. 530 nm), mScarlet-I (ex. 560 nm, em. 600 nm), and FRET (ex. 490 nm, em. 600 nm) was measured 18 hours
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          after IPTG induction (0.05 mM) and normalized to cell count (OD<sub>600</sub>).
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          Statistical significance was determined with Ordinary two-way ANOVA withŠidák's multiple comparison test, with
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          a single pooled variance. *p &lt; 0.05, ****p &lt; 0.001. Data is depicted as mean (n=3) ± SD</i>
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      </div>
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    </div>
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</div>
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</html>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 07:14, 30 September 2024


No part name specified with partinfo tag.

This part can be used for the display of nanobodies on the surface of E.coli. Salema, V. 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 part 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 C-terminus is exposed to the extracellular milieu and contains a myc tag (EQKLISEED) that can be used for purification or detection of full-length protein.

FRET_tetR-Oct1
Figure 3: Fluorescence measurement of mNeonGreen, mScarlet-I and FRET. Fluorescence intensity of mNeonGreen (ex. 490 nm, em. 530 nm), mScarlet-I (ex. 560 nm, em. 600 nm), and FRET (ex. 490 nm, em. 600 nm) was measured 18 hours after IPTG induction (0.05 mM) and normalized to cell count (OD600). Statistical significance was determined with Ordinary two-way ANOVA withŠidák's multiple comparison test, with a single pooled variance. *p < 0.05, ****p < 0.001. Data is depicted as mean (n=3) ± SD

Sequence and Features No part name specified with partinfo tag.


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Functional Parameters

No part name specified with partinfo tag.