Difference between revisions of "Team:ZJU-China/Project/Downstream"

Line 1: Line 1:
 
 
<html lang="zh-CN">
 
<html lang="zh-CN">
  <head>
+
<head>
 
+
 
+
<link href="https://2017.igem.org/Zju/css/bootstrap?action=raw&ctype=text/css" rel="stylesheet" media="screen" />
+
<link href="https://2017.igem.org/Zju/css/bootstraptheme?action=raw&ctype=text/css" rel="stylesheet" id="bs-theme-stylesheet">
+
<link href='https://2017.igem.org/Team:ZJU-China/cs/font?action=raw&ctype=text/css' rel='stylesheet' type='text/css'>
+
<link href="https://2017.igem.org/Zju/css/docs/min?action=raw&ctype=text/css" rel="stylesheet" type="text/css"/>
+
<link href="https://2017.igem.org/Zju/css/doc-style?action=raw&ctype=text/css" rel="stylesheet">
+
<link href="https://2017.igem.org/Zju/css/responsive/min?action=raw&ctype=text/css" rel="stylesheet">
+
  
<script>
+
    <link href="https://2017.igem.org/Zju/css/bootstrap?action=raw&ctype=text/css" rel="stylesheet" media="screen" />
  var _hmt = _hmt || [];
+
    <link href="https://2017.igem.org/Zju/css/bootstraptheme?action=raw&ctype=text/css" rel="stylesheet" id="bs-theme-stylesheet">
</script>
+
    <link href="https://2017.igem.org/Zju/css/docs/min?action=raw&ctype=text/css" rel="stylesheet" type="text/css"/>
  </head>
+
    <!-- <link href="css/doc-style.css" rel="stylesheet" type="text/css"/> -->
 +
    <link href="https://2017.igem.org/Zju/css/doc-style?action=raw&ctype=text/css" rel="stylesheet">
 +
    <link href="https://2017.igem.org/Zju/css/responsive/min?action=raw&ctype=text/css" rel="stylesheet">
 +
    <!-- 按钮的字体 -->
 +
    <link href='https://2017.igem.org/Team:ZJU-China/cs/font?action=raw&ctype=text/css' rel='stylesheet' type='text/css'>
 +
    <!-- 按钮的字体end -->
 +
    <script>
 +
        var _hmt = _hmt || [];
 +
    </script>
 +
    <script src="https://static.igem.org/mediawiki/2017/8/85/ZJUChina_js_docsmin.txt?action=raw&ctype=text/javascript"></script>
 +
</head>
  
  
 
<style>
 
<style>
/*Override default CSS*/
+
    /*Override default CSS*/
#sideMenu, #top_title {display:none;}
+
    #sideMenu, #top_title {display:none;}
#content { padding:0px; width:100%; margin-top:-7px; margin-left:0px;background-color: transparent;}
+
    #content { padding:0px; width:100%; margin-top:-7px; margin-left:0px;background-color: transparent;}
  
  
/* set all basic objects to neutral formatting*/
+
    /* set all basic objects to neutral formatting*/
html, body, div, span, applet, object, iframe, h1, h2, h3, h4, h5, h6, p, blockquote, pre, a, abbr, acronym, address, big, cite, code, del, dfn, em, font, img, ins, kbd, q, s, samp, small, strike, strong, sub, sup, tt, var, b, u, i, center, dl, dt, dd, ol, ul, li, fieldset, form, label, legend, caption {
+
    html, body, div, span, applet, object, iframe, h1, h2, h3, h4, h5, h6, p, blockquote, pre, a, abbr, acronym, address, big, cite, code, del, dfn, em, font, img, ins, kbd, q, s, samp, small, strike, strong, sub, sup, tt, var, b, u, i, center, dl, dt, dd, ol, ul, li, fieldset, form, label, legend, caption {
    margin: 0;
+
        margin: 0;
    padding: 0;
+
        padding: 0;
    border: 0;
+
        border: 0;
    outline: 0;
+
        outline: 0;
    font-size: 100%;
+
        font-size: 100%;
    vertical-align: baseline;
+
        vertical-align: baseline;
    background: transparent;
+
        background: transparent;
}
+
    }
  
#globalWrapper {
+
    #globalWrapper {
    margin: 0;
+
        margin: 0;
    padding: 0;
+
        padding: 0;
    border: 0;
+
        border: 0;
    outline: 0;
+
        outline: 0;
    font-size: 100%;
+
        font-size: 100%;
    vertical-align: baseline;
+
        vertical-align: baseline;
    background: transparent;
+
        background: transparent;
}
+
    }
  
  
/* set page to fill window*/
+
    /* set page to fill window*/
html{
+
    html{
  height: 100%;
+
        height: 100%;
}
+
    }
body {
+
    body {
  min-height: 100%;
+
        min-height: 100%;
  background-color: #FFFFFF;  
+
        background-color: #FFFFFF;
}
+
    }
  
  
#content_wrapper {
+
    #content_wrapper {
    position: relative;
+
        position: relative;
  top: 0px;
+
        top: 0px;
  left: 0px;
+
        left: 0px;
  margin: 0px;
+
        margin: 0px;
    width: 100%;  
+
        width: 100%;
    height: 100%;  
+
        height: 100%;
  text-align:center;
+
        text-align:center;
    background-color: #FFFFFF;
+
        background-color: #FFFFFF;
}
+
    }
  
#content_wrapper p {
+
    #content_wrapper p {
    font-family: 'Tempus Sans ITC',Tahoma, Geneva, sans-serif;
+
        font-family: 'Yanone Kaffeesatz',Tahoma, Geneva, sans-serif;
}
+
    }
  
#content_wrapper a {  
+
    #content_wrapper a {
    color:#004789;
+
        color:#004789;
}
+
    }
  
  
#footer {  
+
    #footer {
    position:absolute;  
+
        position:absolute;
    bottom: 0px;
+
        bottom: 0px;
    left: 0px;
+
        left: 0px;
    width: 100%;
+
        width: 100%;
    margin-top: 0px;
+
        margin-top: 0px;
    margin-bottom: 20px;
+
        margin-bottom: 20px;
    background-color: #FFFFFF;
+
        background-color: #FFFFFF;
    text-align: center;
+
        text-align: center;
}
+
    }
  
.center {  
+
    .center {
  width: 60%;  
+
        width: 60%;
    margin-top: 16px;
+
        margin-top: 16px;
  min-width: 500px;
+
        min-width: 500px;
    padding-bottom:32px;
+
        padding-bottom:32px;
  display: inline-block;      
+
        display: inline-block;
}
+
    }
  
.TextContent {
+
    .TextContent {
  text-align: left;
+
        text-align: left;
  vertical-align: baseline;
+
        vertical-align: baseline;
  font-size: 0.875em;
+
        font-size: 0.875em;
  font-family:'Tempus Sans ITC', verdana, sans-serif;
+
        font-family:'Yanone Kaffeesatz', verdana, sans-serif;
}
+
    }
  
.TextContent h1{  
+
    .TextContent h1{
  text-align: center;
+
        text-align: center;
  font-size: 1.5rem;
+
        font-size: 1.5rem;
  margin-bottom: 1em;
+
        margin-bottom: 1em;
  margin-top: 0.2em;
+
        margin-top: 0.2em;
  color: #3399ff;
+
        color: #3399ff;
  font-family:'Tempus Sans ITC', verdana, sans-serif;
+
        font-family:'Yanone Kaffeesatz', verdana, sans-serif;
}
+
    }
  
.TextContent h2{
+
    .TextContent h2{
  line-height: 1.6;
+
        line-height: 1.6;
  margin-bottom: 0em;
+
        margin-bottom: 0em;
  margin-top: 1em;
+
        margin-top: 1em;
  text-align: left;
+
        text-align: left;
  font-family:'Tempus Sans ITC', verdana, sans-serif;
+
        font-family:'Yanone Kaffeesatz', verdana, sans-serif;
}
+
    }
  
.TextContent p{
+
    .TextContent p{
  margin-bottom: 1.2em;
+
        margin-bottom: 1.2em;
  font-family:'Tempus Sans ITC', verdana, sans-serif;
+
        font-family:'Yanone Kaffeesatz', verdana, sans-serif;
}
+
    }
  
.Page_Wide {
+
    .Page_Wide {
  width:100%;  
+
        width:100%;
}
+
    }
  
.Quarter_Image {
+
    .Quarter_Image {
  width:25%;
+
        width:25%;
  max-height:250px;
+
        max-height:250px;
  float:left;
+
        float:left;
}
+
    }
  
.Quarter_Image img {
+
    .Quarter_Image img {
    display:block;
+
        display:block;
    margin:auto;
+
        margin:auto;
    max-height: 250px;
+
        max-height: 250px;
    width: auto;
+
        width: auto;
}
+
    }
  
.SocialMediaIconWrapper {
+
    .SocialMediaIconWrapper {
    width:100%;
+
        width:100%;
}
+
    }
  
.SocialMediaIcon {
+
    .SocialMediaIcon {
    width:4%;
+
        width:4%;
    height:auto;
+
        height:auto;
    margin-left:20px;
+
        margin-left:20px;
    display: inline-block;
+
        display: inline-block;
    float:left;
+
        float:left;
  
}
+
    }
  
.SocialMediaIcon img{
+
    .SocialMediaIcon img{
    max-width:100%;
+
        max-width:100%;
  height:auto
+
        height:auto
}
+
    }
  
.TeamImage {
+
    .TeamImage {
  width:100%;  
+
        width:100%;
}
+
    }
  
.TeamImage img {
+
    .TeamImage img {
    max-width:100%;
+
        max-width:100%;
  height:auto
+
        height:auto
}
+
    }
  
.sponsors {
+
    .sponsors {
  width: 100%;
+
        width: 100%;
}
+
    }
  
.sponsor_row {  
+
    .sponsor_row {
  width: 100%;
+
        width: 100%;
}
+
    }
  
.sponsor_img {  
+
    .sponsor_img {
  width: 16.6666667%;  
+
        width: 16.6666667%;
  float:left;  
+
        float:left;
}
+
    }
  
.sponsor_img img{  
+
    .sponsor_img img{
  max-width:100%;
+
        max-width:100%;
  height:auto
+
        height:auto
}
+
    }
  
.clear {
+
    .clear {
    clear:both;
+
        clear:both;
}
+
    }
  
.clear.extra_space {
+
    .clear.extra_space {
    height: 20px;
+
        height: 20px;
}
+
    }
  
.line_divider {
+
    .line_divider {
    border-top: 1px solid #d3d3d3;
+
        border-top: 1px solid #d3d3d3;
    width: 98%;
+
        width: 98%;
    margin: auto;
+
        margin: auto;
}
+
    }
  
.copyright{
+
    .copyright{
  color: #888888;
+
        color: #888888;
  font-size: 13px;
+
        font-size: 13px;
  text-align: center !important;
+
        text-align: center !important;
}
+
    }
  
  
.copyright a{
+
    .copyright a{
  color: #d2ebe3;
+
        color: #d2ebe3;
  border-bottom: dotted 1px #a5d8c7;
+
        border-bottom: dotted 1px #a5d8c7;
}
+
    }
  
.PP {
+
    .PP {
    font-family: georgia;
+
        font-family: georgia;
    font-size: 20px;
+
        font-size: 20px;
    font-weight: 100;
+
        font-weight: 100;
    text-align: left;
+
        text-align: left;
}
+
        line-height: 35px;
 +
    }
  
/*文中自定义*/
+
    /*文中自定义*/
.CuteButton:active {
+
    .CuteButton:active {
    -webkit-box-shadow: 0px 3px 0px rgba(0,52,63,1), 0px 3px 6px rgba(0,0,0,.9);
+
        color:white !important;
    -moz-box-shadow: 0px 3px 0px rgba(0,52,63,1), 0px 3px 6px rgba(0,0,0,.9);
+
        -webkit-box-shadow: 0px 3px 0px rgba(0,52,63,1), 0px 3px 6px rgba(0,0,0,.9);
    box-shadow: 0px 3px 0px rgba(0,52,63,1), 0px 3px 6px rgba(0,0,0,.9);
+
        -moz-box-shadow: 0px 3px 0px rgba(0,52,63,1), 0px 3px 6px rgba(0,0,0,.9);
    position: relative;
+
        box-shadow: 0px 3px 0px rgba(0,52,63,1), 0px 3px 6px rgba(0,0,0,.9);
    top: 6px;
+
        position: relative;
}  
+
        top: 6px;
 +
    }
  
.YellowCB {
+
    .CuteButton:visited {
    background-color: rgba(254,196,62,1);
+
        color:white !important;
    -webkit-box-shadow: 0px 9px 0px rgba(245,253,202,1), 0px 9px 25px rgba(0,0,0,.7);
+
        -webkit-box-shadow: 0px 3px 0px rgba(0,52,63,1), 0px 3px 6px rgba(0,0,0,.9);
    -moz-box-shadow: 0px 9px 0px rgba(245,253,202,1), 0px 9px 25px rgba(0,0,0,.7);
+
        -moz-box-shadow: 0px 3px 0px rgba(0,52,63,1), 0px 3px 6px rgba(0,0,0,.9);
    box-shadow: 0px 9px 0px rgba(245,253,202,1), 0px 9px 25px rgba(0,0,0,.7);
+
        box-shadow: 0px 3px 0px rgba(0,52,63,1), 0px 3px 6px rgba(0,0,0,.9);
}
+
        position: relative;
 +
        top: 6px;
 +
    }
  
.GreenCB {
+
    .YellowCB {
    background-color: rgba(25,148,117,1);
+
        background-color: rgba(254,196,62,1);
    -webkit-box-shadow: 0px 9px 0px rgba(4,77,34,1), 0px 9px 25px rgba(0,0,0,.7);
+
        -webkit-box-shadow: 0px 9px 0px rgba(245,253,202,1), 0px 9px 25px rgba(0,0,0,.7);
    -moz-box-shadow: 0px 9px 0px rgba(4,77,34,1), 0px 9px 25px rgba(0,0,0,.7);
+
        -moz-box-shadow: 0px 9px 0px rgba(245,253,202,1), 0px 9px 25px rgba(0,0,0,.7);
    box-shadow: 0px 9px 0px rgba(4,77,34,1), 0px 9px 25px rgba(0,0,0,.7);
+
        box-shadow: 0px 9px 0px rgba(245,253,202,1), 0px 9px 25px rgba(0,0,0,.7);
}
+
    }
  
.SmallCB {
+
    .GreenCB {
    font-size: 1.5em;
+
        background-color: rgba(25,148,117,1);
    margin-left: 2px;
+
        -webkit-box-shadow: 0px 9px 0px rgba(4,77,34,1), 0px 9px 25px rgba(0,0,0,.7);
    padding: 2px 10px ;
+
        -moz-box-shadow: 0px 9px 0px rgba(4,77,34,1), 0px 9px 25px rgba(0,0,0,.7);
    width: 100px;
+
        box-shadow: 0px 9px 0px rgba(4,77,34,1), 0px 9px 25px rgba(0,0,0,.7);
}
+
    }
  
 +
    .SmallCB {
 +
        font-size: 1.5em;
 +
        margin-left: 2px;
 +
        padding: 2px 10px ;
 +
        width: 100px;
 +
    }
  
.ArticleHead{
 
  font-family: 'Tempus Sans ITC',georgia !important;
 
  font-size: 40px !important;
 
  font-weight: 800 !important;
 
  color: #56A36C !important;
 
  text-align: center !important;
 
    margin-bottom: 0 !important;
 
    /*margin-top: .6em !important;*/
 
}
 
  
.H3Head{
 
  font-family: 'Tempus Sans ITC',georgia !important;
 
  color: #FEC43E !important;
 
  font-size: 25px !important;
 
  font-weight: 400 !important;
 
  text-align: left !important;
 
    margin-bottom: 0 !important;
 
    margin-top:.6em !important;
 
}
 
  
.PP{
+
    .ArticleHead{
  font-family:Tahoma, Geneva, sans-serif !important;
+
        font-family: 'Tempus Sans ITC',georgia !important;
  font-size: 20px !important;
+
        font-size: 40px !important;
  font-weight: 100 !important;
+
        font-weight: 800 !important;
  text-align: left !important;
+
        color: #56A36C !important;
}
+
        text-align: center !important;
 +
        margin-bottom: 0 !important;
 +
        margin-top: .6em !important;
 +
    }
  
 +
    .H3Head{
 +
        font-family: 'Tempus Sans ITC',georgia !important;
 +
        color: #FEC43E !important;
 +
        font-size: 25px !important;
 +
        font-weight: 400 !important;
 +
        text-align: left !important;
 +
        margin-bottom: 0 !important;
 +
        margin-top:.6em !important;
 +
    }
  
.Retract{
+
    .PP{
  padding-left: 50px;
+
        font-family:Tahoma, Geneva, sans-serif !important;
}
+
        font-size: 20px !important;
 +
        font-weight: 100 !important;
 +
        text-align: left !important;
 +
    }
  
.GreenAH{
 
  color: #56A36C;
 
}
 
  
.YellowAH{
+
    .Retract{
  color: #FEC43E;
+
        padding-left: 50px;
}
+
    }
  
.contentLi{
+
    .GreenAH{
    margin:0;
+
        color: #56A36C;
    font-size:17px;
+
    }
    line-height: 27px;
+
    padding-bottom: 20px;
+
    color: #111111;
+
}
+
  
.greyTable
+
    .YellowAH{
{
+
        color: #FEC43E;
  color: #7f7f7f;
+
    }
}
+
  
.yellowTable
+
    .contentLi{
{
+
        margin:0;
  color: #CD9B1D;
+
        font-size:17px;
}
+
        line-height: 27px;
 +
        padding-bottom: 20px;
 +
        color: #111111;
 +
    }
  
h2{
+
    .grayTable
  font-family: 'Tempus Sans ITC';
+
    {
  font-weight: 500;
+
        background-color: #979797;
  font-size: 35px;
+
    }
  text-align: left;
+
}
+
  
h3{
+
    .yellowTable
  font-family: 'Tempus Sans ITC';
+
    {
  font-weight: 500;
+
        background-color: #CD9B1D;
  font-size: 32px;
+
        color: #000;
  text-align: left;
+
    }
}
+
  
h4{
+
    h2{
  font-family: 'Tempus Sans ITC';
+
        font-family: 'Tempus Sans ITC';
  font-weight: 500;
+
        font-weight: 500;
  font-size: 28px;
+
        font-size: 35px;
  text-align: left;
+
        text-align: left;
}
+
    }
  
h5{
+
    h3{
  font-family: 'Tempus Sans ITC';
+
        font-family: 'Tempus Sans ITC';
  font-weight: 500;
+
        font-weight: 500;
  font-size: 25px;
+
        font-size: 32px;
  text-align: left;
+
        text-align: left;
}
+
    }
  
p{
+
    h4{
  margin:0;
+
        font-family: 'Tempus Sans ITC';
  font-size:17px;
+
        font-weight: 500;
  line-height: 30px;
+
        font-size: 28px;
  /*padding-bottom: 20px;*/
+
        text-align: left;
  color: #111111;
+
    }
}
+
  
.bs-docs-sidenav
+
    h5{
{
+
        font-family: 'Tempus Sans ITC';
    font-family:Tahoma, Geneva, sans-serif !important;
+
        font-weight: 500;
    font-size: 20px !important;
+
        font-size: 25px;
    font-weight: 100 !important;
+
        text-align: left;
    text-align: left !important;
+
    }
}
+
  
 +
    p{
 +
        margin:0;
 +
        font-size:17px;
 +
        line-height: 30px;
 +
        /*padding-bottom: 20px;*/
 +
        color: #111111;
 +
    }
  
.textli li{
+
    .bs-docs-sidenav
     font-family:Tahoma, Geneva, sans-serif !important;
+
     {
    font-size: 20px !important;
+
        font-family:Tahoma, Geneva, sans-serif !important;
    font-weight: 100 !important;
+
        font-size: 20px !important;
    text-align: left !important;
+
        font-weight: 100 !important;
}
+
        text-align: left !important;
 +
    }
  
.nonStyleTable
 
{
 
    background: transparent !important;
 
    border:none !important;
 
}
 
  
.textimg{
+
    .textli li{
    /*width:50%;*/
+
        font-family:Tahoma, Geneva, sans-serif !important;
    /*margin-top: 20px;*/
+
        font-size: 20px !important;
    /*padding-bottom: 20px;*/
+
        font-weight: 100 !important;
    text-align: center;
+
        text-align: left !important;
}
+
    }
  
 +
    .textimg
 +
    {
 +
        width:60% !important;
 +
    }
  
 +
    .ref
 +
    {
 +
        font-size: 16px !important;
 +
        line-height:22px !important;
 +
        margin-bottom: 5px !important;
 +
        color: #353535;
 +
    }
 +
    /*文中自定义*/
 +
</style>
  
/*老姜要好看一点的表格*/
+
<body id="page-top" data-spy="scroll" data-target=".navbar-fixed-top">
.tableNice thead, .tableNice tr {
+
<div class="page-loader"></div>
    text-align: center;
+
<!-- Docs master nav -->
    border-top-width: 1px;
+
<!-- <h1><a class="navbar-brand" href="index.html">MuMei Lab</a></h1> -->
    border-top-style: solid;
+
<div class="container">
    border-top-color: #b1c6d9;
+
     <!-- header -->
}
+
     <div class="header-w3layouts">
.tableNice {
+
        <!-- Navigation -->
    width: auto;
+
        <nav class="navbar navbar-default navbar-fixed-top">
     margin-left: 30%;
+
            <div class="navbar-header page-scroll">
     border-bottom-width: 1px;
+
                <button type="button" class="navbar-toggle" data-toggle="collapse" data-target=".navbar-ex1-collapse">
    border-bottom-style: solid;
+
                </button>
    border-bottom-color: #D0E9FF;
+
}
+
  
/* Padding and font style */
+
                <a href="https://2017.igem.org/Team:ZJU-China">
.tableNice td, .tableNice th {
+
                    <img style="margin-top:11px" class="navbar-brand"  src="https://static.igem.org/mediawiki/2017/d/d5/ZJUChina_logo.png">
    padding: 5px 10px;
+
                </a>
    font-size: 12px;
+
  
    color: #00343F;
+
                <!-- <h1><a class="navbar-brand" href="index.html">My Design</a></h1> -->
}
+
  
/* Alternating background colors */
+
            </div>
.tableNice tr:nth-child(even) {
+
            <!-- Collect the nav links, forms, and other content for toggling -->
    background: transparent;
+
            <div class="collapse navbar-collapse navbar-ex1-collapse">
 +
                <ul class="nav navbar-nav navbar-right cl-effect-15">
 +
                    <!-- Hidden li included to remove active class from about link when scrolled up past about section -->
 +
                    <li class="hidden"><a class="page-scroll" href="#page-top"></a> </li>
  
}
+
                    <li class="m_nav_item dropdown">
.tableNice tr:nth-child(odd) {
+
                        <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Overview<b class="caret"></b></a>
    background: #EBFFFF;
+
                        <ul class="dropdown-menu ">
}
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Overview">Project Description</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Achievements">Achievements</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:ZJU-China/InterLab">InterLab</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Improve">Improve Parts</a></li>
 +
                        </ul>
 +
                    </li>
  
.tableOuter
+
                    <li class="m_nav_item dropdown">
{
+
                        <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Project<b class="caret"></b></a>
    align: center;
+
                        <ul class="dropdown-menu ">
    text-align: left;
+
                            <!--<li><a href="https://2017.igem.org/Team:ZJU-China/Project">Project Home</a></li>-->
}
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Project/tp">Trichoderma Proof</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Project/voc">VOC sensors</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Project/st">Signal Transduction</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Project/Downstream">Downstream</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Project/conclusion">Conclusion</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Notebook">Notebook</a></li>
 +
                        </ul>
 +
                    </li>
  
.ref
+
                    <li class="m_nav_item dropdown" >
{
+
                        <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Model<b class="caret"></b></a>
    font-size: 16px !important;
+
                        <ul class="dropdown-menu ">
    line-height:22px !important;
+
                            <!--<li><a href="https://2017.igem.org/Team:ZJU-China/Model">Summery</a></li>-->
    margin-bottom: 5px !important;
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Model">VOC analysis</a></li>
    color: #353535;
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Model/Coculture">Coculture</a></li>
}
+
/*老姜要好看一点的表格止*/
+
/*文中自定义*/
+
</style>
+
  
  <body id="page-top" data-spy="scroll" data-target=".navbar-fixed-top">
+
                        </ul>
  <div class="page-loader"></div>
+
                    </li>
      <!-- Docs master nav -->
+
                  <!-- <h1><a class="navbar-brand" href="index.html">MuMei Lab</a></h1> -->
+
  
  <div class="container">
+
                    <li class="m_nav_item dropdown">
      <!-- header -->
+
                        <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Parts<b class="caret"></b></a>
      <div class="header-w3layouts">
+
                        <ul class="dropdown-menu ">
          <!-- Navigation -->
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Parts">All Parts</a></li>
          <nav class="navbar navbar-default navbar-fixed-top">
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Basic_Part">Basic Parts</a></li>
              <div class="navbar-header page-scroll">
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Composite_Part">Composite Parts</a></li>
                  <button type="button" class="navbar-toggle" data-toggle="collapse" data-target=".navbar-ex1-collapse">
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Part_Collection">Part Collection</a></li>
                  </button>
+
                        </ul>
 +
                    </li>
  
                  <a href="https://2017.igem.org/Team:ZJU-China">
+
                    <li><a href="https://2017.igem.org/Team:ZJU-China/Hardware">Hardware</a></li>
                      <img style="margin-top:11px" class="navbar-brand"  src="https://static.igem.org/mediawiki/2017/d/d5/ZJUChina_logo.png">
+
                  </a>
+
  
                  <!-- <h1><a class="navbar-brand" href="index.html">My Design</a></h1> -->
+
                    <li class="m_nav_item dropdown" >
 +
                        <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Safety<b class="caret"></b></a>
 +
                        <ul class="dropdown-menu ">
 +
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Safety">Environment</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Safety/Lab">Laboratory</a></li>
 +
                        </ul>
 +
                    </li>
  
              </div>
+
                    <li class="m_nav_item dropdown">
              <!-- Collect the nav links, forms, and other content for toggling -->
+
                        <a href="#" class="dropdown-toggle link" data-toggle="dropdown">HP<b class="caret"></b></a>
              <div class="collapse navbar-collapse navbar-ex1-collapse">
+
                        <ul class="dropdown-menu ">
                  <ul class="nav navbar-nav navbar-right cl-effect-15">
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Human_Practices">Summary</a></li>
                      <!-- Hidden li included to remove active class from about link when scrolled up past about section -->
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/HP/Silver">Silver</a></li>
                      <li class="hidden"><a class="page-scroll" href="#page-top"></a> </li>
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/HP/Gold_Integrated">Gold</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Engagement">Engagement</a></li>
 +
                        </ul>
 +
                    </li>
  
                      <li class="m_nav_item dropdown">
+
                    <li class="m_nav_item dropdown" >
                          <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Overview<b class="caret"></b></a>
+
                        <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Team<b class="caret"></b></a>
                          <ul class="dropdown-menu ">
+
                        <ul class="dropdown-menu ">
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Overview">Project Description</a></li>
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Team">Teammates</a></li>
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Achievements">Achievements</a></li>
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Attributions">Attribution</a></li>
                              <li><a href="https://2017.igem.org/Team:ZJU-China/InterLab">InterLab</a></li>
+
                            <li><a href="https://2017.igem.org/Team:ZJU-China/Collaborations">Collaboration</a></li>
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Improve">Improve Parts</a></li>
+
                        </ul>
                          </ul>
+
                    </li>
                      </li>
+
  
                      <li class="m_nav_item dropdown">
+
                </ul>
                          <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Project<b class="caret"></b></a>
+
            </div>
                          <ul class="dropdown-menu ">
+
        </nav>
                              <!--<li><a href="https://2017.igem.org/Team:ZJU-China/Project">Project Home</a></li>-->
+
    </div>
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Project/tp">Trichoderma Proof</a></li>
+
</div>
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Project/voc">VOC sensors</a></li>
+
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Project/st">Signal Transduction</a></li>
+
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Project/Downstream">Downstream</a></li>
+
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Project/conclusion">Conclusion</a></li>
+
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Notebook">Notebook</a></li>
+
                          </ul>
+
                      </li>
+
  
                      <li class="m_nav_item dropdown" >
+
<p></p>
                          <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Model<b class="caret"></b></a>
+
<!--<div style="margin-top:0;padding-top:0">-->
                          <ul class="dropdown-menu ">
+
<!--<img id="partsPage" src="https://static.igem.org/mediawiki/2017/b/be/ZJUChina_interlab_banner.jpg" width="100%">-->
                              <!--<li><a href="https://2017.igem.org/Team:ZJU-China/Model">Summery</a></li>-->
+
<!--&lt;!&ndash; <img src="images/HP_temp/banner.jpg" width="100%"> &ndash;&gt;-->
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Model">VOC analysis</a></li>
+
<!--</div>-->
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Model/Coculture">Coculture</a></li>
+
  
                          </ul>
 
                      </li>
 
  
                      <li class="m_nav_item dropdown">
 
                          <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Parts<b class="caret"></b></a>
 
                          <ul class="dropdown-menu ">
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Parts">All Parts</a></li>
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Basic_Part">Basic Parts</a></li>
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Composite_Part">Composite Parts</a></li>
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Part_Collection">Part Collection</a></li>
 
                          </ul>
 
                      </li>
 
 
                      <li><a href="https://2017.igem.org/Team:ZJU-China/Hardware">Hardware</a></li>
 
 
                      <li class="m_nav_item dropdown" >
 
                          <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Safety<b class="caret"></b></a>
 
                          <ul class="dropdown-menu ">
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Safety">Environment</a></li>
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Safety/Lab">Laboratory</a></li>
 
                          </ul>
 
                      </li>
 
 
                      <li class="m_nav_item dropdown">
 
                          <a href="#" class="dropdown-toggle link" data-toggle="dropdown">HP<b class="caret"></b></a>
 
                          <ul class="dropdown-menu ">
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Human_Practices">Summary</a></li>
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/HP/Silver">Silver</a></li>
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/HP/Gold_Integrated">Gold</a></li>
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Engagement">Engagement</a></li>
 
                          </ul>
 
                      </li>
 
 
                      <li class="m_nav_item dropdown" >
 
                          <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Team<b class="caret"></b></a>
 
                          <ul class="dropdown-menu ">
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Team">Teammates</a></li>
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Attributions">Attribution</a></li>
 
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Collaborations">Collaboration</a></li>
 
                          </ul>
 
                      </li>
 
 
                  </ul>
 
              </div>
 
          </nav>
 
      </div>
 
  </div>
 
 
  <p></p>
 
<div style="margin-top:0;padding-top:0">
 
  <img src="https://static.igem.org/mediawiki/2017/0/0a/ZJU_China_home_banner.jpg" width="100%">
 
  <!-- <img src="images/HP_temp/banner.jpg" width="100%"> -->
 
</div>
 
  
  
 
<div style="width: 100%" class="container zjuContent">
 
<div style="width: 100%" class="container zjuContent">
  <div class="col-md-3"></div>
+
    <div class="col-md-3"></div>
 
     <div class="col-md-9" role="main">
 
     <div class="col-md-9" role="main">
      <div class="bs-docs-section">
+
        <p class="bs-docs-section">
  
          <h1 id="design" class="page-header ArticleHead GreenAH">Downstream</h1>
+
            <div style="height: 50px;"></div>
 +
        <h1 id="humanpractice" class="page-header ArticleHead GreenAH">Downstream</h1>
 +
        <h2 class="H2Head" id="introduction">Introduction</h2>
 +
        <p class="PP">When our <em>T.atroviride</em> is activated by the signals which have been described above, they will produce the corresponding effects to save our little plants. Those can be zwittermycin A, chitinase, serine protease and anything you need to protect your lovely plants. Zwittermycin A is an antibiotic which can inhibite the growth of P. nicotianae. Chitinase is a lytic enzyme that breaks down fungal cell walls. Serine protases plays an important role in hydrolyzing the eggshell of root-knot nematodes. With these fungal growth inhibitors our engineered <em>T.atroviride</em> will be able to better protect our plants. </p>
 +
        <div class="imgdiv"><img class="textimg" src="https://static.igem.org/mediawiki/2017/8/82/ZJU_China_Downstream_1.png"></div>
 +
        <h2 class="H2Head" id="zwa">Zwittermycin A</h2>
 +
        <p class="PP">Zwittermicin A is an antibiotic that  has the potential to suppress plant disease due to its broad spectrum activity against certain gram positive and gram negative prokaryotic micro-organisms. Since <em>T.atroviride</em> does not produce Zwittermycin A by itself, a gene cluster obtained from  Bacillus cereus UW85 was introduced into <em>T.atroviride</em>. The genes responsible for the production of zwittermicin A are located on a 16 kb cluster containing nine orfs, from orf1 to orf9, and a self resistant gene zmaR, a gene that encodes an acylation enzyme that deactivate zwittermicin A.<sup>[1]</sup></p>
 +
        <div class="imgdiv"><img class="textimg" src="https://static.igem.org/mediawiki/2017/4/4f/ZJU_China_Downstream_2.png"></div>
 +
        <br><br>
 +
        <div class="imgdiv"><img class="textimg" src="https://static.igem.org/mediawiki/2017/c/c5/ZJU_China_Downstream_3.png"></div>
  
          <h2 id="introduction" class="H2Head">Introduction</h2>
+
        <h2 class="H2Head" id="sp">Serine protease</h2>
                  <p class="PP">When our <em>T.atroviride</em> is activated by the signals which have been described above, they will produce the corresponding effects to save our little plants. Those can be zwittermycin A, chitinase, serine protease and anything you need to protect your lovely plants. Zwittermycin A is an antibiotic which can inhibite the growth of <em>P. nicotianae</em>. Chitinase is a lytic enzyme that breaks down fungal cell walls. Serine protases plays an important role in hydrolyzing the eggshell of root-knot nematodes. With these fungal growth inhibitors our engineered <em>T.atroviride</em> will be able to better protect our plants.</p>
+
        <p class="PP">Root-knot nematodes (Meloidogyne spp.), which are one of the most destructive nematodes, cause the loss of crop about 10%, serious as high as 75%. <sup>[2]</sup> In the present, the egg-parasitic fungus P.lilacinum is the main biocontrol material of root-knot nematodes.P.lilacinum. secretes protease and chitinase to hydrolyze the nematode eggshell, so that the root knot nematodes cannot grow normally.<sup>[3]</sup> However, because P.lilacinum can live in the cornea, the usage of P.lilacinum is still dangerous. Therefore, in this part, the purpose of our project is to give our harmless <em>T.atroviride</em> the ability to kill the root-knot nematodes by overexpressing serine protease which plays an important role in hydrolyzing the eggshell of root-knot nematodes.</p>
                  <div class="imgdiv"><img class="textimg" src="https://static.igem.org/mediawiki/2017/8/82/ZJU_China_Downstream_1.png"></div>
+
                  <p class="capture">Fig.1 Genetic circuit of the downstream</p>
+
  
              <h3 id="za" class="H3Head">Zwittermicin A</h3>
+
        <h2 class="H2Head" id="chi">Chitinase</h2>
                  <p class="PP">Zwittermicin A is an antibiotic that  has the potential to suppress plant disease due to its broad spectrum activity against certain gram positive and gram negative prokaryotic micro-organisms. Since <em>T.atroviride</em> does not produce Zwittermycin A by itself, a gene cluster obtained from  Bacillus cereus UW85 was introduced into <em>T.atroviride</em>. The genes responsible for the production of zwittermicin A are located on a 16 kb cluster containing nine orfs, from orf1 to orf9, and a self resistant gene zmaR, a gene that encodes an acylation enzyme that deactivate zwittermicin A.[1]</p>
+
        <p class="PP">Chitinase is a hydrolytic enzyme that breaks down hydrolytic bonds in chitin. As chitin is a component of the cell walls of fungi and exoskeletal elements of some animals (including worms and arthropods), chitinase has been shown to be useful in biological control against fungi.<sup>[4]</sup> Therefore, in order to inhibit fungl growth, our <em>T.atroviride</em> can produce chitinase when the plants are infested by fungi and the signal conversion systems work well.</p>
                  <div class="imgdiv"><img class="textimg" src="https://static.igem.org/mediawiki/2017/4/4f/ZJU_China_Downstream_2.png"></div>
+
                  <p class="capture">Fig.2A Gene organization of the Zwittermicin A biosynthetic cluster</p>
+
                  <p class="capture">  </p>
+
                  <div class="imgdiv"><img class="textimg" src="https://static.igem.org/mediawiki/2017/0/0b/ZJU_China_Design3.png"></div>
+
                  <p class="capture">Fig.2B Units used in Zwittermicin A Production.[2]</p>
+
  
 +
        <h2 class="H2Head" id="else">Anything else</h2>
 +
        <p class="PP">Your lovely plants will face many challenges in the complex and dangerous soil condition, so that, the little plants must be protected by the strong <em>T.atroviride</em> which can product corresponding effects to inhibite the fungi. The following table can help you choose the right inhibitor to help your plants.</p>
  
              <h3 id="sp" class="H3Head">Serine protease</h3>
+
        <table class="table">
                 <p class="PP">Root-knot nematodes (Meloidogyne spp.), which are one of the most destructive nematodes, cause the loss of crop about 10%, serious as high as 75%. [3] In the present, the egg-parasitic fungus P.lilacinum is the main biocontrol material of root-knot nematodes.P.lilacinum. secretes protease and chitinase to hydrolyze the nematode eggshell, so that the root knot nematodes cannot grow normally.[4] However, because P.lilacinum can live in the cornea, the usage of P.lilacinum is still dangerous. Therefore, in this part, the purpose of our project is to give our harmless <em>T.atroviride</em> the ability to kill the root-knot nematodes by overexpressing serine protease which plays an important role in hydrolyzing the eggshell of root-knot nematodes.</p>
+
            <tr>
              <h3 id="ch" class="H3Head">Chitinase</h3>
+
                 <td class="yellowTable">Inhibitor</td>
                 <p class="PP">Chitinase is a hydrolytic enzyme that breaks down hydrolytic bonds in chitin.  As chitin is a component of the cell walls of fungi and exoskeletal elements of some animals (including worms and arthropods), chitinase has been shown to be useful in biological control against fungi.[5] Therefore, in order to inhibit fungl growth, our <em>T.atroviride</em> can produce chitinase when the plants are infested by fungi and the signal conversion systems work well.</p>
+
                <td class="yellowTable">Part</td>
              <h3 id="anythingelse" class="H3Head">Anything else</h3>
+
                 <td class="yellowTable">Function</td>
                <p class="PP">Your lovely plants will face many challenges in the complex and dangerous soil condition, so that, the little plants must be protected by the strong <em>T.atroviride</em> which can product corresponding effects to inhibite the fungi. The following table can help you choose the right inhibitor to help your plants.</p>
+
            </tr>
  
 +
            <tr>
 +
                <td class="grayTable">dimethyldisulfide(DMDS) and dimethyltrisufide(DMTS)</td>
 +
                <td>BBa_K1493300</td>
 +
                <td>DMTS was shown to have an inhibitory effect on F. oxysporum. DMDS is used as plant growth promoter and at the same time also has been shown a slight inhibition to <em>F. oxysporum</em>.</td>
 +
            </tr>
  
          <h2 id="result" class="H2Head">Result</h2>
+
            <tr>
                    <p class="PP">Because of lacking of time, we only did the experience of serine protease. We structured two plasimads: one could work in the yeast and the other could work in <em>T.atroviride</em>.</p>
+
                <td class="grayTable">Omega-hexatoxin-hv1a</td>
 +
                <td>BBa_K1974001</td>
 +
                <td>Hv1a can bind on insect voltage-gated Calcium channels (CaV1) in the central nervous system, making it paralyze and die eventually</td>
 +
            </tr>
 +
            <tr>
 +
                <td class="grayTable">μ-segestritoxin-Sf1a</td>
 +
                <td>BBa_K1974003</td>
 +
                <td>OAIP can bind on the voltage-gated sodium channel in the insect’s nervous system, making it paralyze and die eventually.</td>
 +
            </tr>
 +
            <tr>
 +
                <td class="grayTable">Orally Active Insecticidal Peptide (OAIP)</td>
 +
                <td>BBa_K1974003</td>
 +
                <td>OAIP can bind on the voltage-gated sodium channel in the insect's nervous system, making it paralyze and die eventually.</td>
 +
            </tr>
 +
            <tr>
 +
                <td class="grayTable">Art-175</td>
 +
                <td>BBa_K1659000</td>
 +
                <td>Art-175 is a fusion protein that kills Gram-negative bacteria, such as Pseudomonas aeruginosa by means of bypassing their outer membranes and catalysing the hydrolysis their cell walls.</td>
 +
            </tr>
 +
            <tr>
 +
                <td class="grayTable">Microcin S</td>
 +
                <td>BBa_K1659100</td>
 +
                <td>tdMicrocin S (MccS) is a narrow-spectrum antibacterial protein that has been shown to exhibit high-potency killing of select strains of E. coli and P. aeruginosa.</td>
 +
            </tr>
  
              <h3 id="yeast" class="H3Head">Yeast</h3>
+
            <tr>
                  <p class="PP">The gene which can express serine protease in yeast was synthesized by Genscript. Before synthesizing this gene, we did codon optimization based on the codon preference of yeast and added a flag-tag to the N-terminal of the serine protease.After extracting the whole proteins of the yeast which transferred plasmid successfully, we performed western-blot and checked the serine protein was expressed in the yeast. (Result is as follow) The band was very shallow, in other words, the concentration of the serine protease was very low.</p>
+
                <td class="grayTable">Plu1537</td>
                  <div class="imgdiv"><img class="textimg" src="https://static.igem.org/mediawiki/2017/d/dd/ZJU_China_Design4.png"></div>
+
                <td>BBa_K1668007</td>
                  <p class="capture">Fig.3 Western-blot result</p>
+
                <td>Plu1537 is a 14kDa insecticidal toxic protein, which has strong toxicity against termites. </td>
                  <p class="PP">The band with red circle was the band of the serine protease and the marker was a protein marker of aidlab.</p>
+
            </tr>
                  <p class="PP">In order to test whether the serine protease could work normally in the yeast, we performed the enzyme activity detection using BAEE solution. We did two sets of experiments: one added PMSF, which was a inhibitor of serine protease, and the other did not. And then, reading the OD253 of these two solutions.(You can know more details about the detection from the protocol) Obviously, the OD253 of the former one is higher than the later one and the values of OD253 increased with time in a period of time ; therefore, we made the conclusion that the yeast produced the serine protase successfully and effectively.</p>
+
        </table>
                  <div class="imgdiv"><img class="textimg" src="https://static.igem.org/mediawiki/2017/f/fb/ZJU_China_Design5.png"></div>
+
                  <p class="capture">Fig.4 The values of OD253 increased with time</p>
+
                  <div class="imgdiv"><img class="textimg" src="https://static.igem.org/mediawiki/2017/7/71/ZJU_China_Design6.png"></div>
+
                  <p class="capture">Fig.5 The values of OD253 increased with time and this solution did not add the PMSF. </p>
+
                  <p class="capture">These were the OD253 of the solutions after a period of reaction.The red one was the solution that did not add the PMSF; the blue one was the solution that added. Obviously, the OD253 of the former one is higher than the later one, so that, we could say that the yeast produced the serine protase successfully and effectively.</p>
+
  
              <h3 id="ta" class="H3Head"><em>T.atroviride</em></h3>
+
        <h2 class="H2Head" id="result">Result</h2>
                  <p class="PP">By contrast to the yeast, the serine protease that worked in the <em>T.atroviride</em> was cloned from the genome of P.lilacinum for they have high homology. Because of lacking of time to extract protein from <em>T.atroviride</em>, we putted the EGFP gene behind the serine protease gene, so that, we could test the serine protease by detecting the fluorescent. From the picture below, mycelium was fluorescent and the <em>T.atroviride</em> expressed the protease successfully. In the future, we will extract the serine protease and detect the activity of it.</p>
+
        <p class="PP">Because of lacking of time, we only did the experience of serine protease. We structured two plasimads: one could work in yeast and the other could work in <em>T.atroviride</em>. Before constructing the plasmid expressed in yeast, we did codon optimization based on the codon preference of  yeast and the gene was synthesized by Genscipt. In the contrast, the serine protease that producted in the <em>T.atroviride</em> was cloned from the genome of P.lilacinum because they have high homology.</p>
                  <div class="imgdiv"><img class="textimg" src="https://static.igem.org/mediawiki/2017/b/ba/ZJU_China_Design7.png"></div>
+
        <hr>
                  <p class="capture">Fig.6 Fluorescence of mycelium</p>
+
        <h2 class="H2Head" id="refer">Reference</h2>
 +
        <p class="PP ref">[1] Stohl E A, Milner J L, Handelsman J. Zwittermicin A biosynthetic cluster[J]. Gene, 1999, 237(2):403-411.</p>
 +
        <p class="PP ref">[2] Wang J P, Wang J X, Liu F, et al. Enhancing the virulence of Paecilomyces lilacinus against Meloidogyne incognita eggs by overexpression of a serine protease. Biotechnology Letters, 32, 1159-1166[J]. Biotechnology Letters, 2010, 32(8):1159-1166.</p>
 +
        <p class="PP ref">[3] Brand D, Roussos S, Pandey A, et al. Development of a bionematicide with Paecilomyces lilacinus to control Meloidogyne incognita.[J]. Applied Biochemistry & Biotechnology, 2004, 118(1-3):81-88.</p>
 +
        <p class="PP ref">[4] Sámi L, Pusztahelyi T, Emri T, et al. Autolysis and aging of Penicillium chrysogenum cultures under carbon starvation: Chitinase production and antifungal effect of allosamidin.[J]. Journal of General & Applied Microbiology, 2001, 47(4):201.</p>
 +
        <br><br><br><br>
 +
    </div>
  
          <h2 id="ref" class="H2Head">Reference</h2>
+
        <!-- 右侧监听开始 -->
              <p class="ref">[1] Stohl E A, Milner J L, Handelsman J. Zwittermicin A biosynthetic cluster[J]. Gene, 1999, 237(2):403-411.</p>
+
        <div class="col-md-3 disappear-on-top" role="complementary">
              <p class="ref">[2] Zwittermicin A, https://en.wikipedia.org/wiki/Zwittermicin_A  ,7 June 2016(24/10/2017)</p>
+
            <nav  style="position: fixed; top: 100px ; left:50px;" class="bs-docs-sidebar hidden-print hidden-xs hidden-sm">
              <p class="ref">[3] Wang J P, Wang J X, Liu F, et al. Enhancing the virulence of Paecilomyces lilacinus against Meloidogyne incognita eggs by overexpression of a serine protease. Biotechnology Letters, 32, 1159-1166[J]. Biotechnology Letters, 2010, 32(8):1159-1166.</p>
+
                <ul class="nav bs-docs-sidenav">
              <p class="ref">[4] Brand D, Roussos S, Pandey A, et al. Development of a bionematicide with Paecilomyces lilacinus to control Meloidogyne incognita.[J]. Applied Biochemistry & Biotechnology, 2004, 118(1-3):81-88.</p>
+
                    <li><a href="#introduction">Introduction</a></li>
              <p class="ref">[5] Sámi L, Pusztahelyi T, Emri T, et al. Autolysis and aging of Penicillium chrysogenum cultures under carbon starvation: Chitinase production and antifungal effect of allosamidin.[J]. Journal of General & Applied Microbiology, 2001, 47(4):201.</p>
+
                    <li><a href="#zwa">Zwittermycin A</a></li>
 +
                    <li><a href="#sp">Serine protease</a></li>
 +
                    <li><a href="#chi">Chitinase</a></li>
 +
                    <li><a href="#else">Anything else</a></li>
 +
                    <li><a href="#result">Result</a></li>
 +
                    <li><a href="#refer">Reference</a></li>
 +
                </ul>
  
 +
                <a class="back-to-top" href="#top">
 +
                    <!--  <i class="fa fa-arrow-up nav_icon"></i> -->
 +
                    <img style="width: 50px;" src="https://static.igem.org/mediawiki/2017/d/da/ZJU_China_UP.png">
 +
                </a>
  
  
 +
            </nav>
 +
        </div>
  
      </div>
+
     </div>
 
+
 
+
 
+
<!-- 右侧监听开始 -->
+
<div class="col-md-3 disappear-on-top" role="complementary">
+
  <nav  style="position: fixed; top: 100px ; left:50px; " class="bs-docs-sidebar hidden-print hidden-xs hidden-sm">
+
     <ul class="nav bs-docs-sidenav">
+
      <li>
+
        <a href="#introduction">Introduction</a>
+
          <ul class="nav">
+
            <li><a href="#za">Zwittermicin A</a></li>
+
            <li><a href="#sp">Serine protease</a></li>
+
            <li><a href="#ch">Chitinase</a></li>
+
            <li><a href="#anythingelse">Anything else</a></li>
+
 
+
          </ul>
+
      </li>
+
 
+
        <li>
+
            <a href="#result">Result</a>
+
            <ul class="nav">
+
                <li><a href="#yeast">Yeast</a></li>
+
                <li><a href="#ta">T.atroviride</a></li>
+
            </ul>
+
        </li>
+
        <li><a href="#ref">Reference</a></li>
+
    </ul>
+
 
+
  <a class="back-to-top" href="#top">
+
  <!--  <i class="fa fa-arrow-up nav_icon"></i> -->
+
    <img style="width: 50px;" src="https://static.igem.org/mediawiki/2017/d/da/ZJU_China_UP.png">
+
  </a>
+
 
+
           
+
          </nav>
+
        </div>
+
       
+
      </div>
+
 
</div>
 
</div>
 
<!-- 右侧监听结束 -->
 
<!-- 右侧监听结束 -->
 
<!-- footer开始 -->
 
<!-- footer开始 -->
 
<div class="footer"  style="background-color: #000000">
 
<div class="footer"  style="background-color: #000000">
  <div class="copyright container text-center">
+
    <div class="copyright container text-center">
      <p><br></p>
+
        <p class="copyright"><br></p>
    <p class="copyright">Copyright &copy; 2017 ZJU-IGEM | <a href="http://www.zju.edu.cn/">Zhejiang University</a> |</p>
+
        <p class="copyright">Copyright &copy; 2017 ZJU-IGEM | <a href="http://www.zju.edu.cn/">Zhejiang University</a> |
    <p><br></p>
+
        </p>
  </div>
+
        <!--<p class="copyright">Contact us:<a class="copyright" href="">WeChat</a></p>-->
 +
        <p class="copyright"><br></p>
 +
    </div>
 
</div>
 
</div>
 
<!-- footer结束 -->
 
<!-- footer结束 -->
Line 647: Line 622:
  
 
<script src="https://code.jquery.com/jquery-2.1.4.min.js"></script>
 
<script src="https://code.jquery.com/jquery-2.1.4.min.js"></script>
  <!--script src="js/bootstrap.min.js"></script-->
+
<!--script src="js/bootstrap.min.js"></script-->
  <script src="https://static.igem.org/mediawiki/2017/8/85/ZJUChina_js_docsmin.txt"></script>
+
<script src="https://static.igem.org/mediawiki/2017/8/85/ZJUChina_js_docsmin.txt"></script>
  <!--script src="js/button.js"></script-->
+
<!--script src="js/button.js"></script-->
  <script src="https://static.igem.org/mediawiki/2017/2/22/ZJUChina_bootstrap.min.txt"></script>
+
<script src="https://static.igem.org/mediawiki/2017/2/22/ZJUChina_bootstrap.min.txt"></script>
  <script src="https://static.igem.org/mediawiki/2017/3/32/ZJUChina_docs.min.txt"></script>
+
<script src="https://static.igem.org/mediawiki/2017/3/32/ZJUChina_docs.min.txt"></script>
  <script src="https://static.igem.org/mediawiki/2017/e/e0/ZJUChina_button.txt"></script>
+
<script src="https://static.igem.org/mediawiki/2017/e/e0/ZJUChina_button.txt"></script>
  
  <!--<script src="js/scrolling-nav.js"></script>-->
+
<script src="https://2017.igem.org/Team:ZJU-China/js/scrollingnavnonPic?action=raw&ctype=text/javascript"></script>
  <script src="https://2017.igem.org/Zju/js/scrollingnav?action=raw&ctype=text/javascript"></script>
+
<!--<script src="https://static.igem.org/mediawiki/2017/9/92/ZJUChina_scrolling-nav.txt"></script>  -->
  <script src="https://static.igem.org/mediawiki/2017/c/c0/ZJUChina_txtSmoothScroll.min.txt"></script>
+
<script src="https://static.igem.org/mediawiki/2017/c/c0/ZJUChina_txtSmoothScroll.min.txt"></script>
  
  </body>
+
</body>
 
</html>
 
</html>

Revision as of 10:14, 25 October 2017

Downstream

Introduction

When our T.atroviride is activated by the signals which have been described above, they will produce the corresponding effects to save our little plants. Those can be zwittermycin A, chitinase, serine protease and anything you need to protect your lovely plants. Zwittermycin A is an antibiotic which can inhibite the growth of P. nicotianae. Chitinase is a lytic enzyme that breaks down fungal cell walls. Serine protases plays an important role in hydrolyzing the eggshell of root-knot nematodes. With these fungal growth inhibitors our engineered T.atroviride will be able to better protect our plants.

Zwittermycin A

Zwittermicin A is an antibiotic that has the potential to suppress plant disease due to its broad spectrum activity against certain gram positive and gram negative prokaryotic micro-organisms. Since T.atroviride does not produce Zwittermycin A by itself, a gene cluster obtained from Bacillus cereus UW85 was introduced into T.atroviride. The genes responsible for the production of zwittermicin A are located on a 16 kb cluster containing nine orfs, from orf1 to orf9, and a self resistant gene zmaR, a gene that encodes an acylation enzyme that deactivate zwittermicin A.[1]



Serine protease

Root-knot nematodes (Meloidogyne spp.), which are one of the most destructive nematodes, cause the loss of crop about 10%, serious as high as 75%. [2] In the present, the egg-parasitic fungus P.lilacinum is the main biocontrol material of root-knot nematodes.P.lilacinum. secretes protease and chitinase to hydrolyze the nematode eggshell, so that the root knot nematodes cannot grow normally.[3] However, because P.lilacinum can live in the cornea, the usage of P.lilacinum is still dangerous. Therefore, in this part, the purpose of our project is to give our harmless T.atroviride the ability to kill the root-knot nematodes by overexpressing serine protease which plays an important role in hydrolyzing the eggshell of root-knot nematodes.

Chitinase

Chitinase is a hydrolytic enzyme that breaks down hydrolytic bonds in chitin. As chitin is a component of the cell walls of fungi and exoskeletal elements of some animals (including worms and arthropods), chitinase has been shown to be useful in biological control against fungi.[4] Therefore, in order to inhibit fungl growth, our T.atroviride can produce chitinase when the plants are infested by fungi and the signal conversion systems work well.

Anything else

Your lovely plants will face many challenges in the complex and dangerous soil condition, so that, the little plants must be protected by the strong T.atroviride which can product corresponding effects to inhibite the fungi. The following table can help you choose the right inhibitor to help your plants.

Inhibitor Part Function
dimethyldisulfide(DMDS) and dimethyltrisufide(DMTS) BBa_K1493300 DMTS was shown to have an inhibitory effect on F. oxysporum. DMDS is used as plant growth promoter and at the same time also has been shown a slight inhibition to F. oxysporum.
Omega-hexatoxin-hv1a BBa_K1974001 Hv1a can bind on insect voltage-gated Calcium channels (CaV1) in the central nervous system, making it paralyze and die eventually
μ-segestritoxin-Sf1a BBa_K1974003 OAIP can bind on the voltage-gated sodium channel in the insect’s nervous system, making it paralyze and die eventually.
Orally Active Insecticidal Peptide (OAIP) BBa_K1974003 OAIP can bind on the voltage-gated sodium channel in the insect's nervous system, making it paralyze and die eventually.
Art-175 BBa_K1659000 Art-175 is a fusion protein that kills Gram-negative bacteria, such as Pseudomonas aeruginosa by means of bypassing their outer membranes and catalysing the hydrolysis their cell walls.
Microcin S BBa_K1659100 tdMicrocin S (MccS) is a narrow-spectrum antibacterial protein that has been shown to exhibit high-potency killing of select strains of E. coli and P. aeruginosa.
Plu1537 BBa_K1668007 Plu1537 is a 14kDa insecticidal toxic protein, which has strong toxicity against termites.

Result

Because of lacking of time, we only did the experience of serine protease. We structured two plasimads: one could work in yeast and the other could work in T.atroviride. Before constructing the plasmid expressed in yeast, we did codon optimization based on the codon preference of yeast and the gene was synthesized by Genscipt. In the contrast, the serine protease that producted in the T.atroviride was cloned from the genome of P.lilacinum because they have high homology.


Reference

[1] Stohl E A, Milner J L, Handelsman J. Zwittermicin A biosynthetic cluster[J]. Gene, 1999, 237(2):403-411.

[2] Wang J P, Wang J X, Liu F, et al. Enhancing the virulence of Paecilomyces lilacinus against Meloidogyne incognita eggs by overexpression of a serine protease. Biotechnology Letters, 32, 1159-1166[J]. Biotechnology Letters, 2010, 32(8):1159-1166.

[3] Brand D, Roussos S, Pandey A, et al. Development of a bionematicide with Paecilomyces lilacinus to control Meloidogyne incognita.[J]. Applied Biochemistry & Biotechnology, 2004, 118(1-3):81-88.

[4] Sámi L, Pusztahelyi T, Emri T, et al. Autolysis and aging of Penicillium chrysogenum cultures under carbon starvation: Chitinase production and antifungal effect of allosamidin.[J]. Journal of General & Applied Microbiology, 2001, 47(4):201.