Difference between revisions of "Team:NTU-LIHPAO-Taiwan/Parts"

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<head>
  
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<script src="https://ajax.googleapis.com/ajax/libs/jquery/1.6.2/jquery.min.js"></script>
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<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
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<title>NTU-LIHPAO-Taiwan</title>
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<style type="text/css">
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/*-------------------------------------------*/
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/*--------------Wiki Page Reset--------------*/
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/*-------------------------------------------*/
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/*-----------------Top icons-----------------*/
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right: 1%;
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#NTU_icon_pic {
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height: 30px;
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width: 69px;
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position: absolute;
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left: auto;
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top: 23px;
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right: 9%;
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bottom: auto;
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z-index: 9999;
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#NTUXLIHPAO_icon {
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height: 40px;
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width: 200px;
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margin:0;
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padding:0;
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position: absolute;
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margin-left: -350px;
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/*background:rgba(100%,100%,100%,0); 主選單色系選擇*/
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left:0;
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top:26px;
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-webkit-transition:background 300ms ease-in-out;
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-moz-transition:background 300ms ease-in-out;
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-o-transition:background 300ms ease-in-out;
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transition:background 300ms ease-in-out;
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line-height:1.1em;
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background-color: rgba(240,240,240,1);
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}
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.Slidemenu ul > li {
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text-decoration:none;
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float: left;
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position: relative;
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height: 27px; /*--改主要選單高度--*/
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}
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.width_small {
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width: 100px; /*--改主要選單寬度--*/
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}
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.Slidemenu ul > li:hover ul.subs {
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width: 150px; /*--改拉出的選單寬度--*/
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border-left-width: 3px;
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border-left-style: solid;
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border-left-color: #3300FF;
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background:rgba(255,255,255,1); /*--改拉出選單顏色--*/
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.Slidemenu ul > li:hover ul li {
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text-decoration:none;
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height: 26px; /*--改拉出的選單高度--*/
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width: 150px; /*--改拉出的選單寬度--*/
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vertical-align: middle;
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-webkit-transition:height 250ms ease-in;
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-moz-transition:height 250ms ease-in;
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-o-transition:height 250ms ease-in;
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transition:height 250ms ease-in;
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background:rgba(0,0,0,0.03);
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left: 5.2%;
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right: auto;
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background: #f7fbff;
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z-index: 5;
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#Content {
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position: absolute;
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top: 60px;
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width: 100%;
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height: auto;
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padding: 0;
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margin: 0;
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transition: 0.5s;
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font-size: 0.8rem;
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font-family: Calibri;
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position: absolute;
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width: 100%;
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top: 0.5rem;
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font-size: 1.5rem;
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text-align: center;
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#Content ul {
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margin: 0.5rem 0rem 0.5rem;
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position: absolute;
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top: 5rem;
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left: 10%;
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width: 90%;
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}
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.main-Content #title {
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color: #ABCDEF;
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cursor: pointer;
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color: #369CF5;
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font-size: 25px;
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padding-top: 5px;
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padding-bottom: 5px;
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line-height: 30px;
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.sub-Content {
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font-size: 18px;
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overflow-y: auto;
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transition: 0.5s;
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width: 85%;
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left: 15%;
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position: relative;
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}
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.sub-Content > li {
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padding-top: 10px;
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padding-bottom: 10px;
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}
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.sub-Content > li a{
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display: block;
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color: #0791DB;
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text-decoration: none;
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transition: 0.5s;
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.sub-Content > li a:hover {
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opacity: 0.5;
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#Healthin_Logo_Content {
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width: 100%;
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height: auto;
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}
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#Healthin_Logo {
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top: 55px;
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padding-left: 3%;
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padding-right: 3%;
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position: absolute;
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width: 94%;
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height: 94%;
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}
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/*-------------------------------------------*/
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/*---------------Introduction----------------*/
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/*-------------------------------------------*/
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.Container_Bottom1 {
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height: 70px;
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width: 100%;
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}
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.Container_Bottom2 {
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height: 70px;
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width: 100%;
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padding-top: 10px;
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.Intro_Picture {
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width: 100%;
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height: auto;
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background-color: #FFF;
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#NTUschool_logo {
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background-color: #FFF;
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padding-left: 9.6%;
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padding-right: 60.4%;
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position: absolute;
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width: 30%;
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height: 100%;
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#LIHPAO_logo {
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background-color: #FFF;
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padding-left: 66.4%;
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padding-right: 9.6%;
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position: absolute;
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width: 24%;
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height: 100%;
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#NTUBST_logo {
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background-color: #FFF;
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padding-left: 49.2%;
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position: absolute;
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width: 7.6%;
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.Text_Sponsor {
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color: #767676;
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font-size: 15px;
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font-family: Calibri;
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text-align: center;
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line-height: 20px;
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}
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/*-------------------------------------------*/
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/*---------------Article Picture-------------*/
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/*-------------------------------------------*/
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.Container_Article_Picture {
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height: 280px;
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width: 100%;
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padding-top: 25px;
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padding-bottom: 25px;
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}
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.Article_Picture {
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width: 480px;
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height: auto;
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left: 50%;
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right: auto;
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background-color: #FFF;
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margin: 0 0 0 -240px;
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position: absolute;
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}
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.Article_PictureText {
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width: 430px;
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height: auto;
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padding-left: 25px;
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padding-right: 25px;
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border-top-width: 2px;
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border-top-style: dashed;
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border-top-color: #fe5838;
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.Container_Article_Picture1 {
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height: 400px;
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width: 100%;
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padding-top: 25px;
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padding-bottom: 25px;
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.Article_Picture1 {
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width: 368px;
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height: auto;
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left: 50%;
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right: auto;
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background-color: #FFF;
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margin: 0 0 0 -184px;
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position: absolute;
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.Article_PictureText1 {
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width: 318px;
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height: auto;
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padding-left: 25px;
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padding-right: 25px;
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border-top-width: 2px;
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border-top-style: dashed;
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border-top-color: #fe5838;
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.Text_Picture {
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font-size: 18px;
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font-family: Calibri;
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text-align: justify;
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line-height: 26px;
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color: #08923a;
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}
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/*-------------------------------------------*/
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/*-----------------Articles------------------*/
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/*-------------------------------------------*/
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#Articles {
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width: 100%;
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height: auto;
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top: -10px;
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position: absolute;
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background-color: #FFF;
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z-index: 1;
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color: #000;
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}
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.ContentBox {
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width: 76%;
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height: auto;
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left: 22%;
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right: 2%;
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position: absolute;
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background-color: #FFF;
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}
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.Text1 {
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font-size: 28px;
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font-family: Calibri;
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left: 20px;
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right: 20px;
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padding-top: 80px;
 +
padding-right: 20px;
 +
padding-bottom: 10px;
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padding-left: 20px;
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border-bottom-width: 2px;
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border-bottom-style: dashed;
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border-bottom-color: #fe5838;
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color: #c00000;
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}
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.Text2 {
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font-size: 24px;
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font-family: Calibri;
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padding-top: 30px;
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padding-right: 50px;
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padding-bottom: 10px;
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padding-left: 50px;
 +
color: #00b050;
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}
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.Text3 {
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font-size: 20px;
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font-family: Calibri;
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text-align: justify;
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padding-top: 10px;
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padding-right: 80px;
 +
padding-bottom: 10px;
 +
padding-left: 80px;
 +
line-height: 26px;
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}
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.Text_TitleUnderline {
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font-size: 22px;
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font-family: Calibri;
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text-align: justify;
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padding-top: 10px;
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padding-bottom: 0px;
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line-height: 26px;
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color: #0070c0;
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text-decoration: underline;
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}
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.Text_italic {
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font-style: italic;
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}
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ol.part1 {
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list-style-type: none;
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margin-left: 0px;
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}
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ol.part2 {
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list-style-type: decimal;
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margin-left: 0px;
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}
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ol.part2 li {
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padding-top: 10px;
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padding-bottom: 10px;
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}
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ol.part3 {
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list-style-type: lower-alpha;
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}
 +
 +
 +
/*-------------------------------------------*/
 +
/*-------------------Media-------------------*/
 +
/*-------------------------------------------*/
 +
 +
@media screen and (min-width: 0px) and (max-width: 1200px) {
 +
 +
#NTUXLIHPAO_icon {
 +
height: 40px;
 +
width: 200px;
 +
position: absolute;
 +
left: 1%;
 +
top: 18px;
 +
right: auto;
 +
bottom: auto;
 +
z-index: 9999;
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}
 +
 +
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#width_small {
 +
width: 95px; /*--改主要選單寬度--*/
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}
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 +
.Slidemenu {
 +
z-index: 9999;
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position: absolute;
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top: 32px;
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bottom: 0px;
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left: 49%;
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right: auto;
 +
list-style:none;
 +
margin:0;
 +
padding:0;
 +
position: absolute;
 +
margin-left: -320px;
 +
/*background:rgba(100%,100%,100%,0); 主選單色系選擇*/
 +
}
 +
}
 +
 +
@media screen and (min-width: 901px) and (max-width: 1050px) {
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.Container_Bottom2 {
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height: 60px;
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width: 100%;
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padding-top: 10px;
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border-top-style: solid;
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border-top-color: #079235;
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border-top-width: 3px;
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}
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}
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@media screen and (min-width: 0px) and (max-width: 900px) {
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 +
.Container_Bottom2 {
 +
height: 50px;
 +
width: 100%;
 +
padding-top: 10px;
 +
border-top-style: solid;
 +
border-top-color: #079235;
 +
border-top-width: 3px;
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}
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}
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</style>
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</head>
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 +
 +
<!---------------------------------------->
 +
<!---------------------------------------->
 +
<!------------------BODY------------------>
 +
<!---------------------------------------->
 +
<!---------------------------------------->
 +
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<body>
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<span class="title">Main Part</span>
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<li><a href="#Second1">Part:BBa_K1841001</a></li>
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<li><a href="#Second2">Part:BBa_K1841004</a></li>
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<li><a href="#Second3">Part:BBa_K1841007</a></li>
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<div class="Text1">Prototype</div>
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<div class="Text2" id="First">Prototype</div>
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<div class="Text3">
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The iGEM Team NTU-LIHPAO-Taiwan 2015 had built a new biological system for the iGEM community. The original design of the system making up the (Product) contains the three elements listed below, in a probiotic – <i>Lactobacillus casei</i> ATCC 393.
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</div>
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<div class="Container_Article_Picture1">
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<div class="Article_Picture1">
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<img src="https://static.igem.org/mediawiki/2015/a/a8/NTU_LASMID.jpg" width="368px"/>
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<div class="Article_PictureText1"><div class="Text_Picture">[Fig.1-1] Designed Plasmid</div></div>
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</div>
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</div>
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<div class="Text1">Main Part</div>
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<div class="Text2" id="Second1">Part: BBa_K1841001</div>
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<div class="Text3"><div class="Text_TitleUnderline">nisI</div></div>
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<div class="Text3">
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Research from Torsten <i>et al.</i> revealed that the highest level of acquired nisin tolerance was achieved after coordinated expression of all four nisin immunity genes, containing <i>nisI</i>, <i>nisF</i>, <i>nisE</i>, and <i>nisG</i>.<a href="#Reference1">[1]</a>  Functional analyses provided evidence that NisI acts as a nisin-sequestering protein and that NisFEG acts as a nisin exporter that expels nisin molecules from the cytoplasmic membrane into the environment.
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</div>
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<div class="Container_Article_Picture">
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<div class="Article_Picture">
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<img src="https://static.igem.org/mediawiki/2015/3/34/NTU-Team-nisI.jpg" width="480px"/>
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<div class="Article_PictureText"><div class="Text_Picture">[Fig.2-1] Nisin Selection</div></div>
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</div>
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</div>
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<div class="Text2" id="Second2">Part: BBa_K1841004</div>
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<div class="Text3"><div class="Text_TitleUnderline">plac promoter</div></div>
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<div class="Text3">
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Lactose operon of <i>Lactobacillus casei</i> In <i>Lactobacillus casei</i> ATCC393 [pLZ15–], the lactose genes are grouped in a cluster transcribed as single operon. The cluster lacTEGF encodes an antiterminator protein (LacT), lactose-specific elements (LacE and LacF) of the phosphotransferase system (PTS) and a phospho-β-galactosidase (LacG).
 +
The promoter region contains a cre element (catabolite responsive element) overlapping the –35 region, which is followed by a highly conserved sequence, the ribonucleic antiterminator (RAT) sequence, and a terminator structure.
 +
The antiterminator activity of LacT is also negatively controlled by glucose, possibly by PTS-mediated phosphorylation as explained below.<a href="#Reference2">[2]</a>
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<div class="Container_Article_Picture">
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<div class="Article_Picture">
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<img src="https://static.igem.org/mediawiki/2015/a/a4/NTU-Team-plac-cl.jpg" width="480px"/>
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<div class="Article_PictureText"><div class="Text_Picture">[Fig.2-2-1] Suicide Sense Part</div></div>
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</div>
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</div>
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<div class="Container_Article_Picture">
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<div class="Article_Picture">
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<img src="https://static.igem.org/mediawiki/2015/9/9a/NTU-Team-nuca.jpg" width="480px"/>
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<div class="Article_PictureText"><div class="Text_Picture">[Fig.2-2-2] Suicide Kill Part</div></div>
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</div>
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</div>
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<div class="Text2" id="Second3">Part: BBa_K1841007</div>
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<div class="Text3"><div class="Text_TitleUnderline">TAT-PYY</div></div>
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<div class="Text3">
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TAT can bring the big molecule substance like DNA, protein, peptide to penetrate the cell. Usually, named as protein transduction domain or membrane transduction domain, it is less than 40 amino acid.<a href="#Reference3">[3]</a> PYY belongs to the gastrointestinal hormones. Once the intestine detects the nutrition, the epidermal cell, L cell of ileum and colon will secrete the PYY. While the blood pass the hypothalamus, PYY will bind to the neuropeptide Y receptor in ventromedial nuclei causing the sense of satiation.<a href="#Reference4">[4]</a>
 +
</div>
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<div class="Container_Article_Picture">
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<div class="Article_Picture">
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<img src="https://static.igem.org/mediawiki/2015/6/6a/NTU-Team-tat-pyy-his.jpg" width="480px"/>
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<div class="Article_PictureText"><div class="Text_Picture">[Fig.2-3] Cell penetrating peptide + PYY</div></div>
 +
</div>
 +
</div>
 +
<div class="Text3">
 +
In this part, we use the constitutive promoter BBa_J23100 to produce signal peptide-PYY-histidine fusion protein. With the His-tag, we can easily purify the signal peptide-PYY from other non-His-tag protein. This part is constructed to test the amount of the PYY that can secret out of the <i>L. casei</i>.
 +
</div>
 +
<div class="Text1">Test Part</div>
 +
<div class="Text2" id="Third1">RFP</div>
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<div class="Container_Article_Picture">
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<div class="Article_Picture">
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<img src="https://static.igem.org/mediawiki/2015/4/4d/NTU-Team-plac-gfp.jpg" width="480px"/>
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<div class="Article_PictureText"><div class="Text_Picture">[Fig.3-1] Suicide sense test</div></div>
 +
</div>
 +
</div>
 +
<div class="Text3">
 +
In this part, we use the RFP protein to testify the function of the lactose operon.
 +
</div>
 +
<div class="Text2" id="Third2">His-PYY</div>
 +
<div class="Container_Article_Picture">
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<div class="Article_Picture">
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<img src="https://static.igem.org/mediawiki/2015/1/12/NTU-Team-his-pyy.jpg" width="480px"/>
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<div class="Article_PictureText"><div class="Text_Picture">[Fig.3-2] PYY</div></div>
 +
</div>
 +
</div>
 +
<div class="Text3">
 +
In this part, we use the constitutive promoter BBa_J23100 to produce histidine-PYY fusion protein. With the His-tag, we can easily purify the PYY from other non-His-tag protein. This part is constructed to test the amount of the PYY under the constitutive promoter in the <i>L. casei</i>.
 +
</div>
 +
<div class="Text2" id="Third3">Signal peptide-PYY-His</div>
 +
<div class="Container_Article_Picture">
 +
<div class="Article_Picture">
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<img src="https://static.igem.org/mediawiki/2015/3/39/NTU-Team-sp-pyy-his.jpg" width="480px"/>
 +
<div class="Article_PictureText"><div class="Text_Picture">[Fig.3-3] Signal peptide + PYY</div></div>
 +
</div>
 +
</div>
 +
<div class="Text3">
 +
In this part, we use the constitutive promoter BBa_J23100 to produce signal peptide-PYY-histidine fusion protein. With the His-tag, we can easily purify the signal peptide-PYY from other non-His-tag protein. This part is constructed to test the amount of the PYY that can secret out of the <i>L. casei</i>.
 +
</div>
 +
<div class="Text2" id="Third4">Y2R</div>
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<div class="Container_Article_Picture">
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<div class="Article_Picture">
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<img src="https://static.igem.org/mediawiki/2015/e/ea/NTU-Team-y2r.jpg" width="480px"/>
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<div class="Article_PictureText"><div class="Text_Picture">[Fig.3-4] Y2R</div></div>
 +
</div>
 +
</div>
 +
<div class="Text3">
 +
In this part, we use the constitutive promoter BBa_J23100 to produce Y2R. This part is constructed to verify the function of PYY.
 +
</div>
 +
<div class="Text1">References</div>
 +
<div class="Text2" id="Fourth">References</div>
 +
<div class="Text3" id="Reference1">
 +
[1] Torsten, S., Stefan, H., Irina, S., and K. D. Entian. Function of Lactococcus lactis Nisin Immunity Genes nisI and nisFEG after Coordinated Expression in the Surrogate Host Bacillus subtilis. The Jurnal of Biological Chemistry, USA. , Vol. 278, pp. 89 -94 (2003)
 +
</div>
 +
<div class="Text3" id="Reference2">
 +
[2] Yu-Kuo Tsai, Hung- Wen Chen, Ta-Chun Lo and Thy-Hou Lin. Specific point mutation in <i>Lactobacillus casei</i> ATCC 27139 cause the phenotype switch from Lac- to Lac+. Microbiology, pp. 751-760(2009)
 +
</div>
 +
<div class="Text3" id="Reference3">
 +
[3] Ju-Chen Cheng, 2009, Investigation on the Characteristic of Heparin-Binding Haemagglutinin 3 Adhesin (HBHA)-Related Peptides and their Transcytosis
 +
</div>
 +
<div class="Text3" id="Reference4">
 +
[4] C. W. le Roux and S. R. Bloom. Peptide YY, appetite and food intake.(2005)
 +
</div>
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Maintained by the iGEM team NTU-LIHPAO-Taiwan&nbsp;&nbsp;&nbsp;&nbsp;©2015 NTU-LIHPAO-Taiwan
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Latest revision as of 13:19, 18 September 2015

NTU-LIHPAO-Taiwan

Prototype
Prototype
The iGEM Team NTU-LIHPAO-Taiwan 2015 had built a new biological system for the iGEM community. The original design of the system making up the (Product) contains the three elements listed below, in a probiotic – Lactobacillus casei ATCC 393.
[Fig.1-1] Designed Plasmid
Main Part
Part: BBa_K1841001
nisI
Research from Torsten et al. revealed that the highest level of acquired nisin tolerance was achieved after coordinated expression of all four nisin immunity genes, containing nisI, nisF, nisE, and nisG.[1] Functional analyses provided evidence that NisI acts as a nisin-sequestering protein and that NisFEG acts as a nisin exporter that expels nisin molecules from the cytoplasmic membrane into the environment.
[Fig.2-1] Nisin Selection
Part: BBa_K1841004
plac promoter
Lactose operon of Lactobacillus casei In Lactobacillus casei ATCC393 [pLZ15–], the lactose genes are grouped in a cluster transcribed as single operon. The cluster lacTEGF encodes an antiterminator protein (LacT), lactose-specific elements (LacE and LacF) of the phosphotransferase system (PTS) and a phospho-β-galactosidase (LacG). The promoter region contains a cre element (catabolite responsive element) overlapping the –35 region, which is followed by a highly conserved sequence, the ribonucleic antiterminator (RAT) sequence, and a terminator structure. The antiterminator activity of LacT is also negatively controlled by glucose, possibly by PTS-mediated phosphorylation as explained below.[2]
[Fig.2-2-1] Suicide Sense Part
[Fig.2-2-2] Suicide Kill Part
Part: BBa_K1841007
TAT-PYY
TAT can bring the big molecule substance like DNA, protein, peptide to penetrate the cell. Usually, named as protein transduction domain or membrane transduction domain, it is less than 40 amino acid.[3] PYY belongs to the gastrointestinal hormones. Once the intestine detects the nutrition, the epidermal cell, L cell of ileum and colon will secrete the PYY. While the blood pass the hypothalamus, PYY will bind to the neuropeptide Y receptor in ventromedial nuclei causing the sense of satiation.[4]
[Fig.2-3] Cell penetrating peptide + PYY
In this part, we use the constitutive promoter BBa_J23100 to produce signal peptide-PYY-histidine fusion protein. With the His-tag, we can easily purify the signal peptide-PYY from other non-His-tag protein. This part is constructed to test the amount of the PYY that can secret out of the L. casei.
Test Part
RFP
[Fig.3-1] Suicide sense test
In this part, we use the RFP protein to testify the function of the lactose operon.
His-PYY
[Fig.3-2] PYY
In this part, we use the constitutive promoter BBa_J23100 to produce histidine-PYY fusion protein. With the His-tag, we can easily purify the PYY from other non-His-tag protein. This part is constructed to test the amount of the PYY under the constitutive promoter in the L. casei.
Signal peptide-PYY-His
[Fig.3-3] Signal peptide + PYY
In this part, we use the constitutive promoter BBa_J23100 to produce signal peptide-PYY-histidine fusion protein. With the His-tag, we can easily purify the signal peptide-PYY from other non-His-tag protein. This part is constructed to test the amount of the PYY that can secret out of the L. casei.
Y2R
[Fig.3-4] Y2R
In this part, we use the constitutive promoter BBa_J23100 to produce Y2R. This part is constructed to verify the function of PYY.
References
References
[1] Torsten, S., Stefan, H., Irina, S., and K. D. Entian. Function of Lactococcus lactis Nisin Immunity Genes nisI and nisFEG after Coordinated Expression in the Surrogate Host Bacillus subtilis. The Jurnal of Biological Chemistry, USA. , Vol. 278, pp. 89 -94 (2003)
[2] Yu-Kuo Tsai, Hung- Wen Chen, Ta-Chun Lo and Thy-Hou Lin. Specific point mutation in Lactobacillus casei ATCC 27139 cause the phenotype switch from Lac- to Lac+. Microbiology, pp. 751-760(2009)
[3] Ju-Chen Cheng, 2009, Investigation on the Characteristic of Heparin-Binding Haemagglutinin 3 Adhesin (HBHA)-Related Peptides and their Transcytosis
[4] C. W. le Roux and S. R. Bloom. Peptide YY, appetite and food intake.(2005)
Maintained by the iGEM team NTU-LIHPAO-Taiwan    ©2015 NTU-LIHPAO-Taiwan