Team:NCTU Formosa/CI design

From 2011.igem.org

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<head>
<head>
<style type = "text/css">
<style type = "text/css">
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#pmenu, #pmenu ul {padding:0; margin:0; list-style-type: none;}
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#pmenu {margin:0px 0px 0px 70px;}
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/*change original background*/
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#pmenu li {float:left;position:relative; list-style-type: none;}
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#pmenu a, #pmenu a:visited {display:block; width:200px; font-size:12px; color:#000; height:28px; line-height:22px; text-decoration:none; text-indent:5px; background:#b2ab9b; border:1px solid #fff; border-width:0 1px 1px 0; text-align:center; list-style-type: none;}
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#top-section{
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#pmenu li:hover > a{background:#dfd7ca; color:#c00;}
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height: 40px;
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#pmenu li ul {display: none;}
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background-color: #cccccc;
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#pmenu li:hover > ul {display:block; position:absolute; top:0; left:201px;}
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border: 0;
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#pmenu > li:hover > ul {left:0; top:24px;}
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}
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</style>  
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img[src*="/wiki/skins/common/images/wiki.png"]{
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    width:0;
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    height:40px;
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    padding-right:975px;
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    background:transparent
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url(https://static.igem.org/mediawiki/2011/0/04/Igem2011.gif) top center no-repeat;
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}
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.firstHeading {
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display:none;
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}
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#search-controls {
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display:none;
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}
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#globalWrapper{
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//background-color: #565659;
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background-image:url(https://static.igem.org/mediawiki/2011/a/ae/Wallpaper2.jpg);
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}
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#content{
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background-color: black;
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border: 0;
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}
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/*top menu*/
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#cm-nav li a.arrow {background-image: url(https://static.igem.org/mediawiki/2011/d/d4/Arrow-right.gif)}
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#cm-nav > li > a.arrow {background-image: url(https://static.igem.org/mediawiki/2011/7/73/Arrow-down.gif)}
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ul#cm-nav ,
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ul#cm-nav ul {
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list-style-type: none;
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margin: 0 0 0 10px;
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list-style-image: none;
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padding: 0px
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}
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ul#cm-nav li {
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position: relative;
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list-style-type: none;
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margin: 0px;
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width: auto;
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float: left;
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list-style-image: none;
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padding: 0px
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}
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ul#cm-nav li li {
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width: auto;
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float: none
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}
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#cm-nav a {
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border-bottom: black 1px solid;
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text-align: left;
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border-left: black 1px solid;
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display: block;
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white-space: nowrap;
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background: #cccccc;
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border-top: black 1px solid;
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border-right: black 1px solid;
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text-decoration: none
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}
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#cm-nav a:focus {
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outline-style: none;
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outline-color: invert;
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outline-width: medium
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}
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#cm-nav a.no-click {cursor: default}
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#cm-nav li a {margin: -1px -1px 0px 0px}
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#cm-nav li li a {margin: 0px 0px -1px}
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#cm-nav > li > a.arrow {padding-right: 14px}
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#cm-nav ul.arrow-pad > li > a {padding-right: 11px}
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#cm-nav li a.arrow ,
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#cm-nav > li > a.arrow {
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background-repeat: no-repeat;
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background-position: right 50%
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}
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#cm-nav li:hover > a ,
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#cm-nav a {color: white}
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#cm-nav li ul ,
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#cm-nav li:hover ul ul ul ul ul ul ul ul ul ul ul ul ul ul ul ul ul ul ul ul ul {
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z-index: 999;
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position: absolute;
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}
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#cm-nav li:hover ul {display: block}
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#cm-nav li li:hover ul ,
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#cm-nav li li li:hover ul ,
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#cm-nav li li li li:hover ul ,
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#cm-nav li li li li li:hover ul ,
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#cm-nav li li li li li li:hover ul ,
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#cm-nav li li li li li li li li li li:hover ul ,
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#cm-nav li li li li li li li li li li li li li li li li:hover ul ,
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#cm-nav li li li li li li li li li li li li li li li li li:hover ul ,
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#cm-nav li li li li li li li li li li li li li li li li li li:hover ul ,
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#cm-nav li li li li li li li li li li li li li li li li li li li:hover ul ,
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#cm-nav li li li li li li li li li li li li li li li li li li li li:hover ul ,
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#cm-nav li li li li li li li li li li li li li li li li li li li li li:hover ul {
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display: block;
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margin-left: 100%
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}
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#cm-nav li:hover ul {margin-left: 0px}
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#cm-nav ul ul {
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top: 1px;
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left: -1px
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}
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#cm-nav li {top: -1px}
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#cm-nav li:hover > a {background-color: #565659}
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#cm-nav a {
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background-color: black;
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border-color: black;
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font-weight: bold;
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font-family: Calibri, Verdana, helvetica, sans-serif;
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font-size: 12pt;
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font-variant: normal;
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padding-left: 10px;
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padding-right: 10px;
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padding-top: 0px;
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padding-bottom: 0px;
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}
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<style type = "text/javascript" language = "JavaScript">
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function out(id_num){
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  var oa = document.getElementById(id_num);
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  if(oa.style.display == "none"){
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    oa.style.display = "block"; 
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  }
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  return false;
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}
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</script>
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/*content*/
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.titleDesign{
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color: white;
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font-family: Calibri, Verdana, helvetica, sans-serif;
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font-weight: bold;
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font-size: 20px;
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font-variant: normal;
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direction:ltr;
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}
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.cont{
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font-size: 14px;
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color: white;
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}
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#banner{    position:relative;
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            margin-left: 10px;
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            background-color: black;
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            height:240px;
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            width: 944px;
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}
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#blueBox {  position:relative;
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            margin-left: 10px;
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            margin-bottom: -5px;
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            margin-top:-3px;
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            background-color: #000066;
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            filter:alpha(opacity=90);opacity:0.9;zoom:1;
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            height:auto;
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            width: 924px;
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}
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#blueBox {  color: white;
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            font-family: Calibri, Verdana, helvetica, sans-serif;
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            font-weight: bold;
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            font-size: 20px;
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            font-variant: normal;
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            direction:ltr;
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            padding-left: 10px;
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            padding-right: 10px;
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}
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#Box1 { position:relative;
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      margin: 0px 0px 0px 10px;
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      background-color: #ede8e2  ;
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      filter:alpha(opacity=95);opacity:0.95;zoom:1;
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      height:auto;
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      width: 924px;                     
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}
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#Box1 { color: black;
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      font-family: Calibri, Verdana, helvetica, sans-serif;
 +
      font-weight: normal;
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      font-size: 12px;
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      font-variant: normal;
 +
      direction:ltr;
 +
      padding: 0 10px 0 10px;
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}
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#Box { position:relative;
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      margin: 0px 0px 0px 10px;
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      background-color: #ede8e2 ;
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      filter:alpha(opacity=98);opacity:0.98;zoom:1;
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      height:auto;
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      width: 864px;                     
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}
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#Box { color: black;
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      font-family: Calibri, Verdana, helvetica, sans-serif;
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      font-size: 18px;
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      font-variant: normal;
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      direction:ltr;
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      padding: 10px 40px 20px 40px;
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      text-align: justify;
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}
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#linkBox {  position:absolute;
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            margin-left: -40px;
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            margin-bottom: -10px;
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            margin-top:-10px;
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            background-color: #000066;
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            filter:alpha(opacity=80);opacity:0.8;zoom:1;
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            height:auto;
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            width: 924px;
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}
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#linkBox {  color: white;
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            font-family: Calibri, Verdana, helvetica, sans-serif;
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            font-weight: bold;
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            font-size: 18px;
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            font-variant: normal;
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            direction:ltr;
 +
            padding-left: 20px;
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}
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#sideBox { position: absolute;
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          top: 304px;
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          left: 720px;
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          height: 975px;
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          width: 240px;
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          background-color: #ede8e2;
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}
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h2{font-family: Calibri, Verdana, helvetica, sans-serif;
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  font-size: 20px;
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  font-weight: bold;
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}
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</style>
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<style>
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table {  
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  border: 0;  
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  font-family: Calibri, Verdana, helvetica, sans-serif;
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  font-size:18px;
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  background-color: #ede8e2;
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  margin: 5px 0 5px 0;
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  padding: 5px 0 5px 10px;
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  text-align:top;
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}
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td{vertical-align:top;}
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h3 {              font-size: 20px;
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                  text-align:right;
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                  text-shadow: #C0C0C0  0px 0px 2px;
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                  vertical-align: baseline;
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            }
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div.intro img {   
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                        border-color: white ;
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border-style:solid;
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                        float:left;
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                        width:expression(this.width>500?"250px":this.width+"px");
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}
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</style>
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</head>
</head>
<body>
<body>
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<div id="banner"><a href = "https://2011.igem.org/Team:NCTU_Formosa"><img src = "https://static.igem.org/mediawiki/2011/8/8c/Banner010.jpg" height="236" width="944"/></a></div>
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 +
<ul id="cm-nav">
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  <li><a href="https://2011.igem.org/Team:NCTU_Formosa">Home</a></li>
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  <li><a onClick="out('cm-nav')" class="arrow">Team </a>
 +
      <ul>
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        <li><a href="https://2011.igem.org/Team:NCTU_Formosa/members">Members</a></li>
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        <li><a href="http://www.nctu.edu.tw/english/index.php">School</a></li>
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        <li><a href="https://2011.igem.org/Team:NCTU_Formosa/gallery">Gallery</a></li>
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      </ul>
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  </li>
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  <li><a onClick="out('cm-nav')" class="arrow">Project</a>
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      <ul class="arrow-pad">
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        <li><a href="https://2011.igem.org/Team:NCTU_Formosa/introduction">Introduction</a></li>
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        <li><a onClick="out('cm-nav')" class="arrow">RNA Thermometer</a>
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            <ul>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/RNA_design">Design</a></li>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/RNA_data">Data</a></li>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/RNA_discussion">Modeling</a></li>
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            </ul>
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        </li>
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        <li><a onClick="out('cm-nav')" class="arrow">CI promoter </a>
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            <ul>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/CI_design">Design</a></li>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/CI_data">Data</a></li>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/CI_discussion">Modeling</a></li>
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            </ul>
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        </li>
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        <li><a onClick="out('cm-nav')" class="arrow">Carotenoid synthesis pathway</a>
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            <ul>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/CSP_design">Design</a></li>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/CSP_data">Data</a></li>
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            </ul>
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        </li>
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        <li><a onClick="out('cm-nav')" class="arrow">Butanol pathway</a>
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            <ul>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/BP_design">Design</a></li>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/BP_data">Data</a></li>
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            </ul>
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        </li>
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        <li><a onClick="out('cm-nav')" class="arrow">Violacein pathway</a>
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            <ul>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/VP_design">Design</a></li>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/VP_data">Data</a></li>
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            </ul>
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        </li>       
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      </ul>
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  </li>
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  <li><a href="https://2011.igem.org/Team:NCTU_Formosa/modeling">Measurements</a></li>
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  <li><a href="https://2011.igem.org/Team:NCTU_Formosa/parts">Parts</a></li>
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  <li><a href="https://2011.igem.org/Team:NCTU_Formosa/safty">Safety</a></li>
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  <li><a href="https://2011.igem.org/Team:NCTU_Formosa/humanpractice">Human Practice</a></li>
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  <li><a href="https://2011.igem.org/Team:NCTU_Formosa/contributions">Attribution</a></li>
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  <li><a onClick="out('cm-nav')" class="arrow">Notebook </a>
 +
      <ul>
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          <li><a onClick="out('cm-nav')" class="arrow">Protocols</a>
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            <ul>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/protocol">Mutation</a></li>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/protocol_F">Flow Cytometry</a></li>
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              <li><a href="https://2011.igem.org/Team:NCTU_Formosa/protocol_G">GC</a></li>
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            </ul>
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          <li><a href="https://2011.igem.org/Team:NCTU_Formosa/calendar">Calendar</a></li>
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      </ul>
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  </li>
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</ul>
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 +
 +
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<br><br>
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<div id="blueBox"><p> High Temperature Induced System – cI Promoter & cI repressor
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</p></div>
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<div id="Box"><h2> Design
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</h2>
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<p>In order to make our target protein has a large amount of performance at high temperature, we use the circuit <a href=""><a href=" http://partsregistry.org/Part:BBa_K098995">BBa_K098995</a>, which is designed by Team Harvard 2008. (Figure1) </p>
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<br>
<br>
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<ul id = "pmenu">
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<center><div><img src = "http://partsregistry.org/wiki/images/9/93/CI-1.png" width="450"></div></center>
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<li><a>Team &#187;</a>
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<br><b>Figure 1.</b><br>
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<ul>
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Part <a href=" http://partsregistry.org/Part:BBa_K098995">BBa_K098995 </a>Design, with constitutive strong promoter <a href=" http://partsregistry.org/Part:BBa_J23114">BBa_J23114 </a>, followed by RBS(Ribosome Binding Site) <a href=" <a href=" http://partsregistry.org/Part:BBa_B0034">BBa_B0034</a>, cI repressor <a href=" http://partsregistry.org/Part:BBa_K098997">BBa_K098997</a>, terminator <a href=" http://partsregistry.org/Part:BBa_B001">BBa_B0014</a>, and high temperature-sensitive cI promoter <a href=" http://partsregistry.org/Part:BBa_R0051">BBa_R0051</a>. Among this circuit, cI repressor(<a href=" http://partsregistry.org/Part:BBa_K098997">BBa_K098997</a>) will be produced constantly to inhibit the cI promoter(<a href=" http://partsregistry.org/Part:BBa_R0051">BBa_R0051</a>) at low temperature. At high temperature, cI repressor(<a href=" http://partsregistry.org/Part:BBa_K098997">BBa_K098997</a>) is still produced, however; cI repressor(<a href=" http://partsregistry.org/Part:BBa_K098997">BBa_K098997</a>) will degrade soon. In this way, cI promoter(<a href=" http://partsregistry.org/Part:BBa_R0051">BBa_R0051</a>) will be activated. This part is designed by 2008 Harvard on backbone pSB1A2, and we transform this part to host strain DH5α for further use.
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<li><a href="https://2011.igem.org/Team:NCTU_Formosa/members">Members</a></li>
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  <br><br>
-
<li><a href="https://2011.igem.org/Team:NCTU_Formosa/school">School</a></li>
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-
<li><a href="https://2011.igem.org/Team:NCTU_Formosa/gallery">Gallery</a></li>
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</ul>
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</li>
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<p>
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<li><a>Project &#187;</a>
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This circuit uses gene <a href="http://partsregistry.org/Part:BBa_K098997">BBa_K098997</a>coding for cI repressor to inhibit the cI promoter <a href=" http://partsregistry.org/Part:BBa_R0051">BBa_R0051</a>. The activity of cI repressor is gradually decreased by elevating temperature from 30 ℃ to 42 ℃. We just need to place our target gene after this circuit  <a href=" http://partsregistry.org/Part:BBa_K098995">BBa_K098995</a>, and then regulate the performance of our target protein by incubating E.coli at different temperatures between 30℃ to 42℃. At higher temperature, our target protein will produce abundantly. On the contrary, at lower temperature, it will not express well. The function of this circuit is illustrated as followed. (Figure 2.) At lower temperature, cI dimer (cI repressor, <a href=" http://partsregistry.org/Part:BBa_K098997">BBa_K098997</a>) will block the RNA polymerase from further transcription, which leads to the gene after this circuit <a href=" http://partsregistry.org/Part:BBa_K098995">BBa_K098995</a> can’t be expressed. Although, at higher temperature, cI dimer (cI repressor, <a href=" http://partsregistry.org/Part:BBa_K098997">BBa_K098997</a>) will degrade and RNA polymerase can transcribe the DNA sequence again, which means that the expression of the gene after this circuit can be recovered.</p>
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<li><a href="https://2011.igem.org/Team:NCTU_Formosa/introduction">Introduction</a></li>
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<li><a>RNA Thermometer &#187;</a>
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<center><div><img src = "http://partsregistry.org/wiki/images/b/bd/CI-2.png" width="700"></div></center>
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<br><b> Figure 2. </b><br>
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  <li><a href="https://2011.igem.org/Team:NCTU_Formosa/RNA_design">Design</a></li>
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First, <a href=" http://partsregistry.org/Part:BBa_K098995">BBa_K098995</a> on backbone PSB1A2 is transformed to host strain DH5α. Then we can find out that, at lower temperature, cI dimer (cI repressor, <a href=" http://partsregistry.org/Part:BBa_K098997">BBa_K098997</a>) will block the RNA polymerase from further transcription, which leads to the gene after this circuit <a href=" http://partsregistry.org/Part:BBa_K098995">BBa_K098995</a> can’t be expressed. Although, at higher temperature, cI dimer (cI repressor, <a href=" http://partsregistry.org/Part:BBa_K098997">BBa_K098997</a>) will degrade and RNA polymerase can transcribe the DNA sequence again, which means that the expression of the gene after this circuit can be recovered. <br><br>
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  <li><a href="https://2011.igem.org/Team:NCTU_Formosa/RNA_data">Data</a></li>
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<a href="https://2011.igem.org/Team:NCTU_Formosa/CI_data" ><font style="Calibri, Verdana, helvetica, sans-serif" color="white" padding-left="10">NEXT >> Data</font>
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<li><a>CI promoter &#187;</a>
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  <li><a href="https://2011.igem.org/Team:NCTU_Formosa/CI_design">Design</a></li>
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Latest revision as of 13:47, 5 October 2011



High Temperature Induced System – cI Promoter & cI repressor

Design

In order to make our target protein has a large amount of performance at high temperature, we use the circuit BBa_K098995, which is designed by Team Harvard 2008. (Figure1)



Figure 1.
Part BBa_K098995 Design, with constitutive strong promoter BBa_J23114 , followed by RBS(Ribosome Binding Site) BBa_B0034, cI repressor BBa_K098997, terminator BBa_B0014, and high temperature-sensitive cI promoter BBa_R0051. Among this circuit, cI repressor(BBa_K098997) will be produced constantly to inhibit the cI promoter(BBa_R0051) at low temperature. At high temperature, cI repressor(BBa_K098997) is still produced, however; cI repressor(BBa_K098997) will degrade soon. In this way, cI promoter(BBa_R0051) will be activated. This part is designed by 2008 Harvard on backbone pSB1A2, and we transform this part to host strain DH5α for further use.

This circuit uses gene BBa_K098997coding for cI repressor to inhibit the cI promoter BBa_R0051. The activity of cI repressor is gradually decreased by elevating temperature from 30 ℃ to 42 ℃. We just need to place our target gene after this circuit BBa_K098995, and then regulate the performance of our target protein by incubating E.coli at different temperatures between 30℃ to 42℃. At higher temperature, our target protein will produce abundantly. On the contrary, at lower temperature, it will not express well. The function of this circuit is illustrated as followed. (Figure 2.) At lower temperature, cI dimer (cI repressor, BBa_K098997) will block the RNA polymerase from further transcription, which leads to the gene after this circuit BBa_K098995 can’t be expressed. Although, at higher temperature, cI dimer (cI repressor, BBa_K098997) will degrade and RNA polymerase can transcribe the DNA sequence again, which means that the expression of the gene after this circuit can be recovered.



Figure 2.
First, BBa_K098995 on backbone PSB1A2 is transformed to host strain DH5α. Then we can find out that, at lower temperature, cI dimer (cI repressor, BBa_K098997) will block the RNA polymerase from further transcription, which leads to the gene after this circuit BBa_K098995 can’t be expressed. Although, at higher temperature, cI dimer (cI repressor, BBa_K098997) will degrade and RNA polymerase can transcribe the DNA sequence again, which means that the expression of the gene after this circuit can be recovered.



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