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| − | Background
| + | Attributions |
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| − | Sortases, enzymes that recognize and cleave the specific sorting signal of secreted proteins to form isopeptide bonds between the secreted proteins and polypeptides, function as protein ligase to form the cell-wall surface of gram-positive bacteria. In case of <i>C.diphtheriae</i>, which belongs to the same genus of our experimental bacteria <i>Corynebacterium glutamicum</i>, has total of 6 sortases, 5 pilus specific sortases (Srt A,B,C,D,E) and 1 housekeeping sortase (Srt F) involved in the formation of all types of pili. <br /> | + | <h1>Software Backend</h1> |
| − | There are 9 types of pillins that comprise pilus; Spa A, B, C, D, E, F, G, H, I, J. SrtA build up SpaA-type pili, which is composed of Spa A, B and C. Srt B/C and Srt d/E each forms SpaD-type pili and SpaH-type pili, which is made up of Spa D, E, F and Spa H, I, J respectively. Like this, the names of each pili types are generally from the names of many Spa proteins. <br />
| + | <h3>Data Collection: Deok ho Kim, Sumin Cho, Seong-ju Lee, Jiwon Kim</h3> |
| − | The picture below shows the process of SpaA type pili formation, using SrtA. Sortase class A enzymes recognize the sequence LPXTG at the carboxyl terminus of surface protein precursors. Cystein of SrtA recognizes LPXTG motif of SpaC, cleaves between T and G, forming SpaC-SrtA intermediate via nucleophilic attack. This intermediate is again attacked by lysine of SrtA bounded SpaA, and the process is continued to form SpaC–SpaAn–SrtA intermediates.<br />
| + | <h3>Pathway Scoring: Jeongsu Hwang, SooWhan Kim, Sumin Cho, Seong-ju Lee, Jiwon Kim</h3> |
| − | Nucleophilic attack by Lysine of SrtF bounded SpaB form SpaC-SpaA(n)-SpaB-SrtF intermediate. The product of this SrtA reaction is covalently linked to lipidⅡ and is then incorporated into the cell wall envelope, terminating the formation of SpaA-type pilus.
| + | <h3>Database Contruction: Sumin Cho, Seong-ju Lee, Jiwon Kim</h3> |
| − | </p>
| + | <h3></h3> |
| − | <img src="https://static.igem.org/mediawiki/2014/thumb/e/ee/Background1.png/800px-Background1.png">
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| − | <p>
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| − | (Architects at the bacterial surface — sortases and the assembly of pili with isopeptide bonds <i>Antoni P. A. Hendrickx, Jonathan M. Budzik, So-Young Oh and Olaf Schneewind</i>)
| + | <h1>Software Frontend</h1> |
| − | </p>
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| − | </div>
| + | <h3>UI & Web Design : Kyeong-hun Jeong, Min ki Choi, Deok ho Kim, Hyeong Woo Kim </h3> |
| − | <div class="title">
| + | <h3>Data visualization : Kyeong-hun Jeong</h3> |
| − | <div class="title_">
| + | <h3>Tutorial & User Guide : Min ki Choi, Hyeong Woo Kim, JuYoung Han, Kyeong-hun Jeong</h3> |
| − | Description
| + | <h3></h3> |
| − | </div>
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| − | <div class="left_line"></div><div class="right_line"></div>
| + | <h1>Wet-lab Work</h1> |
| − | </div>
| + | <h3>Kyujin Jeong, HyeYeong, JuYoung Han, Junhee Kim, Eun San Ko</h3> |
| − | <div class="content_">
| + | <h3></h3> |
| − | <p>
| + | <h1>Wiki</h1> |
| − |
| + | <h3>Editing: Jiwon Kim</h3> |
| − | The main objective of our project is to construct a novel “protein whip” platform, with which we can make <i>Corynebacterium glutamicum</i> to express other corynebacterium’s pili structure comprised of chains of a protein of our choice. As our first try, we decided to make pili made out of green fluorescence proteins (GFP); in order to do so, we substituted SpaA protein, one of the surface proteins in the Pilin A gene cluster, into green fluorescence protein, and transformed a vector containing the modified Pilin A gene cluster into a <i>C. glutamicum</i> strain. <br />Our “protein whip” platform is expected to have many practical applications. For example, pili made out of an enzyme, enzyme whip will enable the reaction to take place with high efficiency, for a great number of the enzyme included in the pili will be able to “attack” the reactants simultaneously. Biofilms made of strains of bacteria that express pili comprised of chains of specific amino acids such as histidine or cysteine that readily bind to heavy metals may be utilized to purify water contaminated with heavy metals. <br />
| + | <h3>Writing: All team members</h3> |
| − | Having a number of potential applications is not the sole merit of our project; by using <i>C. glutamicum</i> instead of widely exploited <i>Escherichia coli</i>, our project also contributes to expanding model organisms used in synthetic biology beyond <i>E. coli</i>.
| + | <h3>Tranlation: JuYoung Han, HyeYeong, Kyujin Jeong, Jiseung Lee</h3> |
| | + | <h3>Web Design and Publishing: Min ki Choi, Kyeong-hun Jeong</h3> |
| | + | <h3></h3> |
| | + | <h1>Funding</h1> |
| | + | <h3>All team members</h3> |
| | + | <h3></h3> |
| | + | <h1>Presentation</h1> |
| | + | <h3>Deok ho Kim, SooWhan Kim, Kyujin Jeong, Jiseung Lee</h3> |
| | + | <h3></h3> |
| | + | <h1>Design</h1> |
| | + | <h3>Hyeong Woo Kim</h3> |
| | + | <h3></h3> |
| | + | <h1>Human Practices</h1> |
| | + | <h3>All team members</h3> |
| | + | <h3></h3> |
| | + | <h1>Instructor</h1> |
| | + | <h3>Byeong-nam Min, Kyoungwoo Jang, and Prof. In Geol Choi</h3> |
| | + | <h3></h3> |
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