Difference between revisions of "Team:NCTU Formosa/Basic Part"

Revision as of 19:51, 18 September 2015

Basic Parts


BBa_K1694002 Lpp-OmpA-NcoI		To display the scFv to the E.coli outer membrane, we chose Lipoprotein-Outer membrane protein A (Lpp-OmpA). According to the paper reference ^[1], we used the first 9 amino acids of Lpp as the signal peptide, and the 46-159 amino acids of OmpA as the anchor. All the sequences is from DH5-alpha strain itself ^[4]. The C-terminal of Lpp-OmpA, located on the outer membrane, is then be fused with the scFv part. In order to change scFv part easily, we provide a basic part which contain NcoI restriction site.
BBa_K1694010 Lpp-OmpA		We also provide Lpp-OmpA without NcoI restriction site.
BBa_K1694003 scFv of anti-VEGF BBa_K1694004 scFv of anti-EGFR BBa_K1694005 scFv of anti-HER2		In our library of customized E.Cotectors, we chose two targeted drugs, Avastin (Bevacizumab, anti-VEGF) ^[1], Erbitux (Cetuximab, anti-EGFR) ^[2] from Drugbank, and a monoclonal antibody, anti-HER2 ^[3]. We select the part of single chain variable fragments (scFv), which is short and will not give too much stress for E.coli expressing it.
BBa_K1694006 GBP BBa_K1694007 FadL-GBP		Gold binding polypeptide (GBP) is a kind of polypeptide which can bind on gold, and is usually used to immobilize proteins on gold surfaces. The mechanism of how GBP binds the gold is not clearly understood, but its polar side-chains, such as those of serine and threonine, seem to interact with gold. We used a 42 amino acids long GBP, which contain 3 repeated amino acid sequences [MHGKTQATSGTIQS]. To display GBP on cell surface, we used long-chain fatty acid transport protein (FadL) as a transmembrane protein, selecting its first 384 amino acids to link with GBP ^[5], signal peptide included.

Reference
[1] C Hartmann et al. (2010) Peptide mimotopes recognized by antibodies cetuximab and matuzumab induce a functionally equivalent anti-EGFR immune response
[2] DrugBank: Bevacizumab (DB00112)
[3] DrugBank: Cetuximab (DB00002)
[4] DrugBank: Trastuzumab (DB00072)
[5] Tae Jung Park et al. (2009) Development of a whole-cell biosensor by cell surface display of a gold-binding polypeptide on the gold surface

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NCTU_Formosa APOllO

National Chaio Tung University
Engineering Building 6 EF455, 1001 University Road, Hsinchu 300, Taiwan, ROC.

@@ Line 186: / Line 186: @@
 <table>
    <tr><td width="20%"></td><td width="13%"></td><td width="67%"></td></tr>
-   <tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694002">BBa_K1694002</a></span><br>Lpp-OmpA-NcoI</td><td></td>
+   <tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694002">BBa_K1694002</a></span><br>Lpp-OmpA-<i>Nco</i>I</td><td></td>
    <td>
    To display the scFv to the <i>E.coli</i> <B>outer membrane</B>, we chose Lipoprotein-Outer membrane protein A (Lpp-OmpA).
 According to the paper reference <sup>[1]</sup>, we used the first 9 amino acids of Lpp as the signal peptide, and the 46-159 amino acids of OmpA as the anchor. All the sequences is from DH5-alpha strain itself <sup>[4]</sup>. <br>
 The C-terminal of Lpp-OmpA, located on the outer membrane, is then be fused with the scFv part.<br>
-   In order to change scFv part easily, we provide a basic part which contain NcoI restriction site.
+   In order to change scFv part easily, we provide a basic part which contain <i>Nco</i>I restriction site.
 </td></tr>
    <tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694010">BBa_K1694010</a></span><br>Lpp-OmpA</td><td></td>
-   <td>We also provide Lpp-OmpA without <i>Nco</i> restriction site.
+   <td>We also provide Lpp-OmpA without <i>Nco</i>I restriction site.
 </td></tr>
    <tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694003">BBa_K1694003</a></span><br>