Difference between revisions of "Team:UofC Calgary/Parts"

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{{UofC_Calgary}}
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{{Team:UofC_Calgary/UofC_Calgary}}
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<html>
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{{Team:UofC_Calgary/assets/plugins/bootstrap-social/bootstrap-social.css}}
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{{Team:UofC_Calgary/assets/plugins/socicon/socicon.css}}
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{{Team:UofC_Calgary/assets/plugins/font-awesome/css/font-awesome.min.css}}
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{{Team:UofC_Calgary/assets/plugins/simple-line-icons/simple-line-icons.min.css}}
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{{Team:UofC_Calgary/assets/plugins/bootstrap/css/bootstrap.min.css}}
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{{Team:UofC_Calgary/assets/plugins/revo-slider/css/settings.css}}
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{{Team:UofC_Calgary/assets/base/css/themes/default.css}}
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{{Team:UofC_Calgary/assets/base/css/custom.css}}
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{{Team:UofC_Calgary/assets/google-fonts.css}}
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{{Team:UofC_Calgary/assets/plugins/jquery.min.js}}
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{{Team:UofC_Calgary/assets/plugins/jquery-migrate.min.js}}
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{{Team:UofC_Calgary/assets/plugins/bootstrap/js/bootstrap.min.js}}
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{{Team:UofC_Calgary/assets/plugins/revo-slider/js/jquery.themepunch.tools.min.js}}
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{{Team:UofC_Calgary/assets/plugins/revo-slider/js/jquery.themepunch.revolution.min.js}}
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{{Team:UofC_Calgary/assets/base/js/components.js}}
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{{Team:UofC_Calgary/assets/base/js/app.js}}
  
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<html lang="en">
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<!--<![endif]-->
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<!-- BEGIN HEAD -->
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<head>
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<meta charset="utf-8"/>
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<title>iGEM Calgary 2016</title>
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<meta http-equiv="X-UA-Compatible" content="IE=edge">
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<meta content="width=device-width, initial-scale=1.0" name="viewport"/>
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<meta http-equiv="Content-type" content="text/html; charset=utf-8">
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<meta content="" name="description"/>
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<meta content="" name="author"/>
  
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<link rel="shortcut icon" href="favicon.ico"/>
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</head>
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<body class="c-layout-header-fixed">
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<!-- BEGIN: LAYOUT/HEADERS/HEADER-1 -->
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<!-- BEGIN: HEADER -->
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<header class="c-layout-header c-layout-header-3 c-layout-header-default-mobile c-header-seethrough" style="padding-top: 10px;
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    top: 0;
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    position: fixed;
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    z-index: 9995;
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    width: 100%;
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">
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<div class="c-navbar">
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<div class="container-fluid" style="padding-left: 0px; padding-right: 0px;">
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<!-- BEGIN: BRAND -->
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<div class="c-navbar-wrapper clearfix">
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<div class="c-brand c-pull-left">
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<a href="https://2016.igem.org/Team:UofC_Calgary" class="c-logo">                                         
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<img src="https://static.igem.org/mediawiki/2016/9/9a/Logo-1.png" alt="iGEM" class="c-desktop-logo">
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<img src="https://static.igem.org/mediawiki/2016/9/9a/Logo-1.png" alt="iGEM" class="c-desktop-logo-inverse">
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<img src="https://static.igem.org/mediawiki/2016/9/9a/Logo-1.png" alt="iGEM" class="c-mobile-logo">
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</a>
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<button class="c-hor-nav-toggler" type="button" data-target=".c-mega-menu">
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<span class="c-line"></span>
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<span class="c-line"></span>
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<span class="c-line"></span>
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</button>
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</div>
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<!-- END: BRAND -->
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<!-- BEGIN: HOR NAV -->
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<!-- BEGIN: LAYOUT/HEADERS/MEGA-MENU -->
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<nav class="c-mega-menu c-pull-right c-mega-menu-dark c-mega-menu-dark-mobile c-fonts-uppercase c-fonts-bold">
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<!-- BEGIN: MEGA MENU -->
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<ul class="nav navbar-nav c-theme-nav">
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<li class="c-menu-type-classic">
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<a href="https://2016.igem.org/Team:UofC_Calgary" class="c-link dropdown-toggle">Home</a>
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</li>
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<li class="c-active c-menu-type-classic">
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<a href="https://2016.igem.org/Team:UofC_Calgary/Description" class="c-link dropdown-toggle">Project</a>
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<ul class="dropdown-menu c-menu-type-inline c-pull-left">
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<li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Description">Problem</a>
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</li>
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<li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Experiments">Our Experiments</a>
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</li>
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                                                                <li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Results">Our Results</a>
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</li>
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                                                                <li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Model">Model</a>
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</li><li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Design">Applied Design</a>
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</li>
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<li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Parts">Parts Registry</a>
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</li>
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<li>
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                                                                <a href="https://2016.igem.org/Team:UofC_Calgary/Composite_Part">Composite Parts</a>
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</li>
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</ul>
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</li>
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<li class="c-menu-type-classic">
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<a href="https://2016.igem.org/Team:UofC_Calgary/Judging" class="c-link">Judging</a>
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</li>
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<li class="c-menu-type-classic">
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<a href="https://2016.igem.org/Team:UofC_Calgary/Human_Practices" class="c-link dropdown-toggle">Human Practices</a>
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<ul class="dropdown-menu c-menu-type-inline c-pull-left">
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<li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Integrated_Practices">Integrated Practices</a>
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</li>
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<li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Collaborations">Collaborations</a>
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</li>
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<li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Engagement">Engagement</a>
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</li>
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<li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Safety">Safety</a>
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</li>
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<li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/HP/Silver">Silver</a>
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</li>
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<li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/HP/Gold">Gold</a>
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</li>
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<li>
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                                    <a href="https://2016.igem.org/Team:UofC_Calgary/Policy"> Policy Brief </a>
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                                    </li>
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</ul>
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</li>
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                                <li class="c-menu-type-classic">
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<a href="https://2016.igem.org/Team:UofC_Calgary/Notebook" class="c-link">Notebook</a>
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</li>
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<li class="c-menu-type-classic">
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<a href="https://2016.igem.org/Team:UofC_Calgary/Team" class="c-link dropdown-toggle">Team</a>
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<ul class="dropdown-menu c-menu-type-inline c-pull-left">
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                                                                <li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Team">Meet Our Team</a>
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</li>
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<li>
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<a href="https://2016.igem.org/Team:UofC_Calgary/Attributions">Attributions</a>
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</li>
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</ul>
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</li>
  
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<li>
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<a href="https://2016.igem.org/Main_Page" data-toggle="modal" data-target="#login-form" class="c-btn-border-opacity-04 c-btn btn-no-focus c-btn-header btn btn-sm c-btn-border-1x c-btn-white c-btn-circle c-btn-sbold"><i class="icon-chemistry"></i> iGEM </a>
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</li>
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<div class="column full_size">
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<li class="c-quick-sidebar-toggler-wrapper">
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<a href="#" class="c-quick-sidebar-toggler">
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<span class="c-line"></span>
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<p>Each team will make new parts during iGEM and will submit them to the Registry of Standard Biological Parts. The iGEM software provides an easy way to present the parts your team has created. The <code>&lt;groupparts&gt;</code> tag (see below) will generate a table with all of the parts that your team adds to your team sandbox.</p>
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<span class="c-line"></span>
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<p>Remember that the goal of proper part documentation is to describe and define a part, so that it can be used without needing to refer to the primary literature. Registry users in future years should be able to read your documentation and be able to use the part successfully. Also, you should provide proper references to acknowledge previous authors and to provide for users who wish to know more.</p>
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<span class="c-line"></span>
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</a>
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</li>
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</ul>
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<!-- END MEGA MENU -->
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</div>
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</div>
 
</div>
 
</div>
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</header>
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<!-- END: HEADER -->
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<!-- BEGIN: LAYOUT/SIDEBARS/QUICK-SIDEBAR -->
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<nav class="c-layout-quick-sidebar">
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<div class="column half_size">
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<div class="c-header">
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<div class="highlight">
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<button type="button" class="c-link c-close">
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<h5>Note</h5>
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<i class="icon-login"></i>
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<p>Note that parts must be documented on the <a href="http://parts.igem.org/Main_Page"> Registry</a>. This page serves to <i>showcase</i> the parts you have made. Future teams and other users and are much more likely to find parts by looking in the Registry than by looking at your team wiki.</p>
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</button>
 
</div>
 
</div>
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<div class="c-content">
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<div class="c-section">
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<h3>Theme Colors</h3>
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<div class="c-settings">
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<span class="c-color c-default c-active" data-color="default"></span>
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<span class="c-color c-green1" data-color="green1"></span>
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<span class="c-color c-green2" data-color="green2"></span>
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<span class="c-color c-green3" data-color="green3"></span>
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<span class="c-color c-yellow1" data-color="yellow1"></span>
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<span class="c-color c-yellow2" data-color="yellow2"></span>
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<span class="c-color c-yellow3" data-color="yellow3"></span>
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<span class="c-color c-red1" data-color="red1"></span>
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<span class="c-color c-red2 " data-color="red2"></span>
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<span class="c-color c-red3" data-color="red3"></span>
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<span class="c-color c-purple1" data-color="purple1"></span>
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<span class="c-color c-purple2" data-color="purple2"></span>
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<span class="c-color c-purple3" data-color="purple3"></span>
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<span class="c-color c-blue1" data-color="blue1"></span>
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<span class="c-color c-blue2" data-color="blue2"></span>
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<span class="c-color c-blue3" data-color="blue3"></span>
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<span class="c-color c-brown1" data-color="brown1"></span>
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<span class="c-color c-brown2" data-color="brown2"></span>
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<span class="c-color c-brown3" data-color="brown3"></span>
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<span class="c-color c-dark1" data-color="dark1"></span>
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<span class="c-color c-dark2" data-color="dark2"></span>
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<span class="c-color c-dark3" data-color="dark3"></span>
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</div>
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</div>
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<div class="c-section">
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<h3>Header Type</h3>
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<div class="c-settings">
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<input type="button" class="c-setting_header-type btn btn-sm c-btn-square c-btn-border-1x c-btn-white c-btn-sbold c-btn-uppercase active" data-value="boxed" value="boxed"/>
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<input type="button" class="c-setting_header-type btn btn-sm c-btn-square c-btn-border-1x c-btn-white c-btn-sbold c-btn-uppercase" data-value="fluid" value="fluid"/>
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</div>
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</div>
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<div class="c-section">
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<h3>Header Mode</h3>
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<div class="c-settings">
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<input type="button" class="c-setting_header-mode btn btn-sm c-btn-square c-btn-border-1x c-btn-white c-btn-sbold c-btn-uppercase active" data-value="fixed" value="fixed"/>
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<input type="button" class="c-setting_header-mode btn btn-sm c-btn-square c-btn-border-1x c-btn-white c-btn-sbold c-btn-uppercase" data-value="static" value="static"/>
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</div>
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</div>
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<div class="c-section">
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<h3>Mega Menu Style</h3>
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<div class="c-settings">
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<input type="button" class="c-setting_megamenu-style btn btn-sm c-btn-square c-btn-border-1x c-btn-white c-btn-sbold c-btn-uppercase active" data-value="dark" value="dark"/>
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<input type="button" class="c-setting_megamenu-style btn btn-sm c-btn-square c-btn-border-1x c-btn-white c-btn-sbold c-btn-uppercase" data-value="light" value="light"/>
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</div>
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</div>
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<div class="c-section">
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<h3>Font Style</h3>
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<div class="c-settings">
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<input type="button" class="c-setting_font-style btn btn-sm c-btn-square c-btn-border-1x c-btn-white c-btn-sbold c-btn-uppercase active" data-value="default" value="default"/>
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<input type="button" class="c-setting_font-style btn btn-sm c-btn-square c-btn-border-1x c-btn-white c-btn-sbold c-btn-uppercase" data-value="light" value="light"/>
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</div>
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</div>
 
</div>
 
</div>
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</nav>
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<!-- END: LAYOUT/SIDEBARS/QUICK-SIDEBAR -->
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<!-- BEGIN: PAGE CONTAINER -->
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<div class="c-layout-page" style="padding-top: 88px;">
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<!-- BEGIN: PAGE CONTENT -->
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<!--BEGIN: SLIDE #1 -->
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            <img alt="" src="https://static.igem.org/mediawiki/2016/f/f5/T--UofC_Calgary--compositepartsbanner.jpg" data-bgposition="center center" data-bgfit="cover" data-bgrepeat="no-repeat" style="width: 100%; height: 100%;">
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<!-- END: LAYOUT/SLIDERS/REVO-SLIDER-1 -->
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<!-- BEGIN: CONTENT/MISC/ABOUT-1 -->
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      <div class="c-content-box c-size-md" style="background-color: #172724">
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  <div class="container">
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            <div class="row">
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                <div class="col-span-12">
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                    <p style="color:#FFFFFF;">The backbone of the U of Calgary project are the composite parts. All of our parts are composite, and are an amalgamation of different registry parts which act together to provide a novel function. The composite parts designed for this project are based around the modified Bowman-Birk Protease Inhibitor (mBBI) and the competency-inducing <a href="http://parts.igem.org/Part:BBa_K1444018"><i>ComK</i> part</a> that was submitted to the registry by the <a href="https://2014.igem.org/Team:Calgary/Notebook/Parts">2014 UofC_Calgary</a> team.</p><br>
  
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<p style="color:#FFFFFF;">A variety of different parts were used across all of the mBBI composite constructs. We used Pveg, a constitutive <i>E. coli</i> and <i>B. subtilis</i> promoter, and double terminators to book-end internal genes upon chromosomal integration. To improve secretion and transdermal delivery of mBBI from <i>B. subtilis</i>, we added a secretion (sec) tag as well as a transdermal (TD1) tag. The secretion tag is a localization tag for the Sec pathway in <i>B. subtilis</i> (Fu <i>et al.,</i> 2007). <a href="http://parts.igem.org/Part:BBa_E0040:Experience">TD1</a> is a part previously developed by the <a href="https://2013.igem.org/Team:USTC_CHINA">USTC_China 2013</a> iGEM team, and it is designed to allow for the trafficking of peptides across the skin.</p><br>
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<p style="color:#FFFFFF;">The second major composite part was the <i>ComK</i> construct. ComK is a transcription factor and master regulator for competency in <i>B. subtilis</i>. We modified the original <a href="http://parts.igem.org/Part:BBa_K1444018"><i>ComK</i></a> sequence, which was first submitted by the <a href="https://2014.igem.org/Team:Calgary/Notebook/Parts">2014 UofC_Calgary</a> team, to remove unwanted restriction sites from within the <i>ComK</i> gene itself. The <i>ComK</i> gene and xylose-inducible promoter were then flanked with sequences homologous to the <i>amyE</i> locus in <i>B. subtilis</i>, a non-essential gene responsible for the degradation of starch. These flanking sequences were added to allow for the integration of a controllable ComK into the <i>B. subtilis</i> genome. The integration of a xylose-inducible ComK allows for the transformation of <i>B. subtilis</i> to be more time-efficient and less resource-intensive. </p><br>
  
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<p style="color:#FFFFFF;">Our last construct does not fall into either of the two previous categories. The construct is an integration cassette, which is a mix of both the mBBI and <i>Comk</i> constructs. Our cassette contains <i>thrC</i> integration sequences which flank the sec and TD1 tag. However, the mBBI sequence is replaced with the restriction sites for BamHI and XmaI restriction enzymes. Addition of the restriction sites allows for directional cloning of any desired insert, allowing <i>B. subtilis</i> to be used as a platform for future biotherapeutics, creating a portable mini-pharmacy.
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</p>
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                </div>
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          </div>
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      </div>
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        <!-- END: CONTENT/MISC/ABOUT-1 -->
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<!-- BEGIN: CONTENT/MISC/ABOUT-1 -->
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<div class="c-content-box c-size-md" style="background-color: #172724; padding-bottom:0px;">
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  <div class="container">
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<div class="c-content-title-1">
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<h3 class="c-center c-font-uppercase c-font-bold" style="color:#EBDEBE">Composite Parts</h3>
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<div class="c-line-center c-theme-bg">
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</div>
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</div>
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              <div class="row">
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                            <div class="col-sm-3" style="padding-left:0px">
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                                  <h3 class="human-practices-div-2">BBa_K2008001
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  </h3>
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  <p class="human-practices-div-2">BBI with N-terminal KSCI solubility tag fused to GFP for expression in <em>E. coli</em></p>
  
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<ul><li><p class="human-practices-div-2">Sequence-confirmed</p></li><li><p class="human-practices-div-2">Submitted to the registry</p></li><li><p class="human-practices-div-2"><a href="http://parts.igem.org/Part:BBa_K2008001">Registry Link</a></p></li></ul>
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                            </div>
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                            <div class="col-sm-3" style="padding-left:7px; padding-right:7px">
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                                  <h3 class="human-practices-div-2">BBa_K2008002
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</h3>
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  <p class="human-practices-div-2">BBI with N-terminal sec and TD1 tags fused to GFP and under the pVeg promoter for expression in <em>B. subtilis</em></p><ul><li><p class="human-practices-div-2">Sequence-confirmed</p></li><li><p class="human-practices-div-2">Submitted to the registry</p></li><li><p class="human-practices-div-2"><a href="http://parts.igem.org/Part:BBa_K2008002">Registry Link</a></p></li></ul>           
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                            </div>
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<div class="col-sm-3" style="padding-left:7px; padding-right:7px">
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                                  <h3 class="human-practices-div-2">BBa_K2008003
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</h3>
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  <p class="human-practices-div-2">BBI with N-terminal KSCI solubility, sec, and TD1 tags fused to GFP and under the pVeg promoter for expression in <em>B. subtilis</em></p><ul><li><p class="human-practices-div-2">Sequence-confirmed</p></li><li><p class="human-practices-div-2">Submitted to the registry</p></li><li><p class="human-practices-div-2"><a href="http://parts.igem.org/Part:BBa_K2008003">Registry Link</a></p></li></ul>             
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                            </div>
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                            <div class="col-sm-3" style="padding-right:0px">   
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                                  <h3 class="human-practices-div-2">BBa_K2008004
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</h3>
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  <p class="human-practices-div-2">BBI with N-terminal sec and TD1 tags under the pVeg promoter for expression in <em>B. subtilis</em></p><ul><li><p class="human-practices-div-2">Sequence-confirmed</p></li><li><p class="human-practices-div-2">Submitted to the registry</p></li><li><p class="human-practices-div-2"><a href="http://parts.igem.org/Part:BBa_K2008004">Registry Link</a></p></li></ul>
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<div class="row">
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          <div class="col-sm-3" style="padding-left:0px">
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            <img src="https://static.igem.org/mediawiki/2016/8/8d/T--UofC_Calgary--BBa_K2008001img.png" width="300px" height="88"px/>
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          </div>
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          <div class="col-sm-3" style="padding-left:7px; padding-right:7px">
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            <img src="https://static.igem.org/mediawiki/2016/4/48/T--UofC_Calgary--BBa_K2008002img.png" width="300px" height="88"px/>
 +
          </div>
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          <div class="col-sm-3" style="padding-left:7px; padding-right:7px">
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            <img src="https://static.igem.org/mediawiki/2016/0/08/T--UofC_Calgary--BBa_K2008003img.png" width="300px" height="88"px/>
 +
          </div>
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          <div class="col-sm-3" style="padding-right:0px">
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            <img src="https://static.igem.org/mediawiki/2016/e/e1/T--UofC_Calgary--BBa_K2008004img.png" width="300px" height="88"px/>     
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          </div>
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<div class="row">
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 +
                                  <h3 class="human-practices-div-2"><font color=#A2310F>♥</font>BBa_K2008005
 +
  </h3>
 +
  <p class="human-practices-div-2">BBI with KSCI solubility, sec, and TD1 tags under the pVeg promoter for expression in <em>B. subtilis</em></p><ul><li><p class="human-practices-div-2">Sequence-confirmed</p></li><li><p class="human-practices-div-2">Characterized</p></li><li><p class="human-practices-div-2">Submitted to the registry</p></li><li><p class="human-practices-div-2"><a href="http://parts.igem.org/Part:BBa_K2008005">Registry Link</a></p></li></ul>
 +
                            </div>
 +
                            <div class="col-sm-3" style="padding-left:7px; padding-right:7px">
 +
                                  <h3 class="human-practices-div-2">BBa_K2008006
 +
 +
</h3>
 +
  <p class="human-practices-div-2">Improved version of <i>comK</i> gene with removal of internal restriction sites</p><ul><li><p class="human-practices-div-2">Sequence-confirmed</p></li><li><p class="human-practices-div-2">Submitted to the registry</p></li><li><p class="human-practices-div-2"><a href="http://parts.igem.org/Part:BBa_K2008006">Registry Link</a></p></li></ul>             
 +
                            </div>
 +
<div class="col-sm-3" style="padding-left:7px; padding-right:7px">
 +
                                  <h3 class="human-practices-div-2"><font color=#A2310F>♥</font>BBa_K2008007
 +
</h3>
 +
  <p class="human-practices-div-2">Improved version of <i>comK</i> with addition of flanking regions of <i>amyE</i> homology</p><ul><li><p class="human-practices-div-2">Sequence-confirmed</p></li><li><p class="human-practices-div-2">Submitted to the registry</p></li><li><p class="human-practices-div-2"><a href="http://parts.igem.org/Part:BBa_K2008007">Registry Link</a></p></li></ul> 
 +
           
 +
                            </div>
 +
                            <div class="col-sm-3" style="padding-right:0px">   
 +
                                  <h3 class="human-practices-div-2"><font color=#A2310F>♥</font>BBa_K2008008
 +
</h3>
 +
  <p class="human-practices-div-2">Modular secretion platform containing sec and TD1 tags with flanking regions of <i>thrC</i> homology for chromosomal integration in <i>B. subtilis</i></p><ul><li><p class="human-practices-div-2">Sequence-confirmed</p></li><li><p class="human-practices-div-2">Submitted to the registry</p></li><li><p class="human-practices-div-2"><a href="http://parts.igem.org/Part:BBa_K2008008">Registry Link</a></p></li></ul>         
 +
                            </div>
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                </div>
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  <div class="row">
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          <div class="col-sm-3" style="padding-left:0px">
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            <img src="https://static.igem.org/mediawiki/2016/f/fb/T--UofC_Calgary--BBa_K2008005img.png" width="300px" height="88"px/>
 +
          </div>
 +
          <div class="col-sm-3" style="padding-left:7px; padding-right:7px">
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          <img src="https://static.igem.org/mediawiki/2016/b/b8/T--UofC_Calgary--BBa_K2008006img.png" width="300px" height="88"px/>
 +
          </div>
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          <div class="col-sm-3" style="padding-left:7px; padding-right:7px">
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            <img src="https://static.igem.org/mediawiki/2016/d/d1/T--UofC_Calgary--BBa_K2008007img.png" width="300px" height="88"px/>
 +
          </div>
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          <div class="col-sm-3" style="padding-right:0px">
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            <img src="https://static.igem.org/mediawiki/2016/e/e5/T--UofC_Calgary--BBa_K2008008img.png" width="300px" height="88"px/>     
 +
          </div>
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        </div>
 +
<br><br><p style="color:#FFFFFF;">References</p><br>
 +
<p style="color:#FFFFFF;">Fu, L.L., Xu, Z.R., Li, W.F., Shuai, J.B., Lu, P., and Hu, C.X. (2007). Protein secretion pathways in <i>Bacillus subtilis</i>: implication for optimization of heterlogous protein secretion. Biotechnology Advances <i>25</i>(1): 1-12.</p>
  
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<p>We encourage teams to start completing documentation for their parts on the Registry as soon as you have it available. The sooner you put up your parts, the better you will remember all the details about your parts. Remember, you don't need to send us the DNA sample before you create an entry for a part on the Registry. (However, you <b>do</b> need to send us the DNA sample before the Jamboree. If you don't send us a DNA sample of a part, that part will not be eligible for awards and medal criteria.)</p>
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<li>Part Name</li>
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<li>Creator</li>
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<li>Short Description (60 characters on what the DNA does)</li>
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We encourage you to put up <em>much more</em> information as you gather it over the summer. If you have images, plots, characterization data and other information, please also put it up on the part page. </p>
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Latest revision as of 01:08, 20 October 2016

iGEM Calgary 2016

The backbone of the U of Calgary project are the composite parts. All of our parts are composite, and are an amalgamation of different registry parts which act together to provide a novel function. The composite parts designed for this project are based around the modified Bowman-Birk Protease Inhibitor (mBBI) and the competency-inducing ComK part that was submitted to the registry by the 2014 UofC_Calgary team.


A variety of different parts were used across all of the mBBI composite constructs. We used Pveg, a constitutive E. coli and B. subtilis promoter, and double terminators to book-end internal genes upon chromosomal integration. To improve secretion and transdermal delivery of mBBI from B. subtilis, we added a secretion (sec) tag as well as a transdermal (TD1) tag. The secretion tag is a localization tag for the Sec pathway in B. subtilis (Fu et al., 2007). TD1 is a part previously developed by the USTC_China 2013 iGEM team, and it is designed to allow for the trafficking of peptides across the skin.


The second major composite part was the ComK construct. ComK is a transcription factor and master regulator for competency in B. subtilis. We modified the original ComK sequence, which was first submitted by the 2014 UofC_Calgary team, to remove unwanted restriction sites from within the ComK gene itself. The ComK gene and xylose-inducible promoter were then flanked with sequences homologous to the amyE locus in B. subtilis, a non-essential gene responsible for the degradation of starch. These flanking sequences were added to allow for the integration of a controllable ComK into the B. subtilis genome. The integration of a xylose-inducible ComK allows for the transformation of B. subtilis to be more time-efficient and less resource-intensive.


Our last construct does not fall into either of the two previous categories. The construct is an integration cassette, which is a mix of both the mBBI and Comk constructs. Our cassette contains thrC integration sequences which flank the sec and TD1 tag. However, the mBBI sequence is replaced with the restriction sites for BamHI and XmaI restriction enzymes. Addition of the restriction sites allows for directional cloning of any desired insert, allowing B. subtilis to be used as a platform for future biotherapeutics, creating a portable mini-pharmacy.

Composite Parts

BBa_K2008001

BBI with N-terminal KSCI solubility tag fused to GFP for expression in E. coli

BBa_K2008002

BBI with N-terminal sec and TD1 tags fused to GFP and under the pVeg promoter for expression in B. subtilis

BBa_K2008003

BBI with N-terminal KSCI solubility, sec, and TD1 tags fused to GFP and under the pVeg promoter for expression in B. subtilis

BBa_K2008004

BBI with N-terminal sec and TD1 tags under the pVeg promoter for expression in B. subtilis

♥BBa_K2008005

BBI with KSCI solubility, sec, and TD1 tags under the pVeg promoter for expression in B. subtilis

  • Sequence-confirmed

  • Characterized

  • Submitted to the registry

  • Registry Link

BBa_K2008006

Improved version of comK gene with removal of internal restriction sites

♥BBa_K2008007

Improved version of comK with addition of flanking regions of amyE homology

♥BBa_K2008008

Modular secretion platform containing sec and TD1 tags with flanking regions of thrC homology for chromosomal integration in B. subtilis



References


Fu, L.L., Xu, Z.R., Li, W.F., Shuai, J.B., Lu, P., and Hu, C.X. (2007). Protein secretion pathways in Bacillus subtilis: implication for optimization of heterlogous protein secretion. Biotechnology Advances 25(1): 1-12.

iGEM

iGEM is an international competition promoting synthetic biology as a means to solve social, economic and humanitarian problems around the globe. The iGEM Jamboree is held in Boston annually. In 2016, over 300 teams are competing against each other.

Latest Entries

Fully Trained!

Our entire team received a full BioSafety education from the University of Calgary! This entailed going to classes to prepare for a final quiz that tested our ability to be safe in the lab. Several of our members also had radiation training and clearance to ensure that work done with radiation was safe!

Read More

Latest Pictures

View More

Find us

Located in Calgary, Alberta, Canada.

  • University of Calgary
  • igem.calgary@gmail.com

Copyright © University of Calgary 2016.

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