Team:Korea U Seoul/Project/sub design

From 2014.igem.org

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<p>Our part is a “pilin A gene cluster”. Pilin A gene cluster is coding for pili A of Corynebacterium diphtheriae. It has four coding regions and one regulatory region. Regulatory region is its own promoter. Four coding regions are sortase A, enzyme that makes pili A and pilin spa A & B & C that are components of pili A. WIth these all regions, we can make pili A. And we are going to modifying this part to generate "Our Favorite Protein Whip", especially spa A<br />
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                We try to express the pearl gene (nacrein) on the <i>E.coli</i>, and check the CaCO3-<i>E.coli</i> structrue. The gene related to nacrein was made by the genetic synthetic technology and go through codon optimization step which for expression of <i>E.coli.</i><br /><br />
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Because of internal restriction enzyme site in our part, we have to slice it to two parts. We can use pilin A gene cluster by cutting and fusing these two parts. Vector is pSB1C3
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pAT vector is the vector which can express the protein on surface of cell. We made vectors containing 4type of nacrein. The promotor of protien expression was pBAD, so we used arabinose for expression.<br /><br />
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Before checking the functioning of Nacrein protein which displayed on cell, purify 4type of nacrein protein and check functioning of them.
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                         <td>BBa_K114600</td>
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                         <td>BBa_K1377000</td>
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                         <td>coding</td>
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                         <td>Device</td>
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                             YfaL_Nacrein(WT)<br />
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                             promoter of pili<br />
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                            wild type Nacrein, not including the signal sequence. Two carbonic anhydrase(CA) domains and Gly-X-Asn repeat(R)
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A gene cluster + spaA front
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                         </td>
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                         <td>2830</td>
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                         <td>1137bp</td>
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                         <td>Areum Goh</td>
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                         <td>Jade Minseob Yoo</td>
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                     </tr>
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                         <td>BBa_K114601</td>
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                         <td>BBa_K1377001</td>
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                         <td>coding</td>
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                         <td>Device</td>
                         <td>
                         <td>
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                             YfaL_Nacrein CA<br />
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                             spaA backward + sortase A + spaB + spaC
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                            C-terminal deletion mutant covering one CA domain, not including the signal sequence
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                         <td>2188</td>
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                         <td>7772bp</td>
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                         <td>Areum Goh</td>
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                         <td>Jade Minseob Yoo</td>
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                        <td>BBa_K114602</td>
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                        <td>coding</td>
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                        <td>
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                            YfaL_Nacrein DR<br />
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                            The Gly-X-Asn deletion domain. Deleted Repeat(DR)
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                        </td>
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                        <td>2563</td>
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                        <td>Areum Goh</td>
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                    </tr>
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                        <td>BBa_K114603</td>
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                        <td>coding</td>
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                        <td>
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                            YfaL_Nacrein R<br />
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                            Gly-X-Asn repeat domain(R)
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                        <td>1807</td>
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                        <td>Areum Goh</td>
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                     </tr>
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                 </table>
                 </table>

Revision as of 05:53, 13 October 2014

Design & Part

Our part is a “pilin A gene cluster”. Pilin A gene cluster is coding for pili A of Corynebacterium diphtheriae. It has four coding regions and one regulatory region. Regulatory region is its own promoter. Four coding regions are sortase A, enzyme that makes pili A and pilin spa A & B & C that are components of pili A. WIth these all regions, we can make pili A. And we are going to modifying this part to generate "Our Favorite Protein Whip", especially spa A
Because of internal restriction enzyme site in our part, we have to slice it to two parts. We can use pilin A gene cluster by cutting and fusing these two parts. Vector is pSB1C3

Name Type Description Length Designer
BBa_K1377000 Device promoter of pili
A gene cluster + spaA front 		1137bp Jade Minseob Yoo
BBa_K1377001 Device spaA backward + sortase A + spaB + spaC 7772bp Jade Minseob Yoo

Retrieved from "http://2014.igem.org/Team:Korea_U_Seoul/Project/sub_design"