Difference between revisions of "Team:TJUSLS China/Proof"

Line 1: Line 1:
 +
 
<html>
 
<html>
 
<head>
 
    <!-- head -->
 
    <link href="https://2016.igem.org/Team:TJUSLS_China/home/home_style?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css" media="all" />
 
    <!--start slider -->
 
    <link href="https://2016.igem.org/Team:TJUSLS_China/home/home_lrtk?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css" media="all" />
 
    <link rel="stylesheet" href="https://2016.igem.org/Team:TJUSLS_China/home/home_fwslider?action=raw&amp;ctype=text/css" media="all">
 
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_lrtk_js?action=raw&ctype=text/javascript"></script>
 
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_jquery_min?action=raw&ctype=text/javascript"></script>
 
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_jquery_min?action=raw&ctype=text/javascript"></script>
 
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_jquery-ui_min?action=raw&ctype=text/javascript"></script>
 
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_fwslider1?action=raw&ctype=text/javascript"></script>
 
    <!--end slider -->
 
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_jquery-hover-effect?action=raw&ctype=text/javascript"></script>
 
    <script type="text/javascript">
 
        //Image Hover
 
        jQuery(document).ready(function(){
 
            jQuery(function() {
 
                jQuery('ul.da-thumbs > li').hoverdir();
 
            });
 
        });
 
    </script>
 
    <!-- Add fancyBox main JS and CSS files -->
 
    <script src="https://2016.igem.org/Team:TJUSLS_China/home/home_jquery.magnific-popup?action=raw&ctype=text/javascript" type="text/javascript"></script>
 
    <link href="https://2016.igem.org/Team:TJUSLS_China/home/home_magnific-popup?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css">
 
    <script>
 
        $(document).ready(function() {
 
            $('.popup-with-zoom-anim').magnificPopup({
 
                type: 'inline',
 
                fixedContentPos: false,
 
                fixedBgPos: true,
 
                overflowY: 'auto',
 
                closeBtnInside: true,
 
                preloader: false,
 
                midClick: true,
 
                removalDelay: 300,
 
                mainClass: 'my-mfp-zoom-in'
 
            });
 
        });
 
    </script>
 
    <!--nav-->
 
    <script>
 
        $(function() {
 
            var pull = $('#pull');
 
            menu = $('nav ul');
 
            menuHeight = menu.height();
 
 
            $(pull).on('click', function(e) {
 
                e.preventDefault();
 
                menu.slideToggle();
 
            });
 
 
            $(window).resize(function(){
 
                var w = $(window).width();
 
                if(w > 320 && menu.is(':hidden')) {
 
                    menu.removeAttr('style');
 
                }
 
            });
 
        });
 
    </script>
 
 
    <!-- origin -->
 
    <script>
 
        buttonClick = {};
 
    </script>
 
    <title>Home</title>
 
    <!--<script src="js/jquery.min.js"></script>-->
 
    <link href="https://2016.igem.org/Team:TJUSLS_China/CSS_Home_bootstr?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css" media="all">
 
    <link href="https://2016.igem.org/Team:TJUSLS_China/CSS_Home_style?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css" media="all" />
 
    <meta name="viewport" content="width=device-width, initial-scale=1">
 
    <meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
 
    <meta name="keywords" content="Solutem  Responsive web template, Bootstrap Web Templates, Flat Web Templates, Andriod Compatible web template,
 
Smartphone Compatible web template, free webdesigns for Nokia, Samsung, LG, SonyErricsson, Motorola web design" />
 
    <script type="application/x-javascript"> addEventListener("load", function() { setTimeout(hideURLbar, 0); }, false); function hideURLbar(){ window.scrollTo(0,1); } </script>
 
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/js/jquery-min?
 
action=raw&ctype=text/javascript"></script>
 
    <!---- start-smoth-scrolling---->
 
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/js/move-top?
 
action=raw&ctype=text/javascript"></script>
 
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/js/easing?
 
action=raw&ctype=text/javascript"></script>
 
    <script type="text/javascript">
 
        jQuery(document).ready(function($) {
 
            $(".scroll").click(function(event){
 
                event.preventDefault();
 
                $('html,body').animate({scrollTop:$(this.hash).offset().top},1200);
 
            });
 
        });
 
    </script>
 
    <!---End-smoth-scrolling---->
 
    <link href="https://2016.igem.org/Team:TJUSLS_China/CSS_Home_component?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css"  />
 
    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/js/modernizr.custom?
 
action=raw&ctype=text/javascript"></script>
 
</head>
 
 
<style>
 
<style>
    #bodyContent h2 {
+
     #content{width:100%;padding:0px;margin:0px;background-color: #ededed;}
        font-size: 2em;
+
    }
+
 
+
    .cbp-spmenu-push-toleft {
+
        left: -115px;
+
    }
+
 
+
     #content {
+
        width: 100%;
+
        padding: 15px;
+
        border: none;
+
        color: black;
+
        margin-left: 20px;
+
        margin-right: auto;
+
        background-color: #ffffff;
+
        position: relative;
+
    }
+
 
     #sideMenu, #top_title
 
     #sideMenu, #top_title
 
     {
 
     {
 
         display: none;
 
         display: none;
 
     }
 
     }
 
+
    body{font: 16px sans-serif;}
     .right10{
+
     .footer_bg .footer {
         right: 10px;
+
         padding: 0;
 +
        border-top: 6px solid #222222;
 
     }
 
     }
     .right190{
+
     .footer_bg .footer_btm {
         right: 190px;
+
         margin-top: 4%;
 +
        border-top: none;
 +
        padding: 2% 4%;
 +
        padding-top: 2%;
 +
        max-width: 1458px;
 +
        margin: 0 auto;
 
     }
 
     }
 
 
</style>
 
</style>
 +
<head>
 +
    <meta charset="UTF-8">
 +
    <title></title>
 +
    <link rel="stylesheet" href="https://2016.igem.org/Team:TJUSLS_China/home/index_style?action=raw&amp;ctype=text/css">
 +
    <script type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/index_js_two?action=raw&ctype=text/javascript"></script>
 +
    <script type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/index_js_one?action=raw&ctype=text/javascript"></script>
 +
</head>
 
<body>
 
<body>
<div class="cbp-spmenu-push" id="body">
+
<!--menu-->
    <div class="header_bg">
+
<div class="header-box">
        <div class="wrap">
+
    <div class="header">
            <div class="header">
+
        <!-- start h_menu4 -->
                <div class="logo">
+
        <div class="h_menu4">
                    <a href="index.html"><img src="images/logo.png" alt="" /></a>
+
            <a class="toggleMenu" href="">Menu</a>
                </div>
+
            <ul id="top_ul_my" class="nav">
                <!-- start h_menu4 -->
+
                <script>
                <div class="h_menu4">
+
                    function getHover(number){
                    <a class="toggleMenu" href="">Menu</a>
+
                        var top_ul_my_lis = $("#top_ul_my").find(">li");
                    <ul id="top_ul_my" class="nav">
+
                        console.log(top_ul_my_lis);
                        <script>
+
                        if(number == 0) top_ul_my_lis[0].setAttribute("class", "active hover");
                            function getHover(number){
+
                        else top_ul_my_lis[0].setAttribute("class", "active");
                                var top_ul_my_lis = $("#top_ul_my").find(">li");
+
                                if(number == 0) top_ul_my_lis[0].setAttribute("class", "active hover");
+
                                else top_ul_my_lis[0].setAttribute("class", "active");
+
  
                                for(var i = 1; i < top_ul_my_lis.length; i++){
+
                        for(var i = 1; i < top_ul_my_lis.length; i++){
                                    if(i == number) {
+
                            if(i == number) {
                                        top_ul_my_lis[i].setAttribute("class", "hover");
+
                                top_ul_my_lis[i].setAttribute("class", "hover");
                                    }else{
+
                            }else{
                                        top_ul_my_lis[i].setAttribute("class", "");
+
                                top_ul_my_lis[i].setAttribute("class", "");
                                    }
+
                                }
+
 
                             }
 
                             }
 +
                        }
 +
                    }
  
                            $(document).ready(function() {
+
                    $(".header").hover(function(event){
                                //console.log("ready");
+
                        event.stopPropagation();
                                var top_ul_my_lis = $("#top_ul_my").find(">li");
+
                        $('#top_ul_my li').attr("class","");
                                for(var i = 1; i < top_ul_my_lis.length; i++) {
+
                    });
                                    $(top_ul_my_lis[i]).mouseover(getHover(i));
+
                                }
+
                            });
+
  
                        </script>
+
                </script>
                      <li class="active"><a class="active" href="https://2016.igem.org/Team:TJUSLS_China/home">TJUSLS</a></li>
+
                <li onmouseover="getHover(0)"><a href="https://2016.igem.org/Team:TJUSLS_China/home">TJUSLS</a></li>
                        <li ><a href="https://2016.igem.org/Team:TJUSLS_China/Project" class="root">project</a>
+
                <li onmouseover="getHover(1)" class=""><a href="" class="root">project</a>
                            <ul>
+
                    <ul>
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Description">Description</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Description">Description</a></li>
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Design">Design</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Design">Design</a></li>
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Experiments">Experiments</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Experiments">Experiments</a></li>
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Proof">Proof of Concept</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Proof">Proof of Concept</a></li>
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Demonstrate">Demonstrate</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Demonstrate">Demonstrate</a></li>
                               
+
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/PROJECT/PROTOCOLS">protocols</a></li>
+
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Notebook">Notebook</a></li>
+
                            </ul>
+
                        </li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Parts">PARTS</a>
+
                            <ul>
+
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Basic_Part">Basic Parts</a></li>
+
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Composite_Part">Composite Parts</a></li>
+
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Part_Collection">Part Collection</a></li>
+
  
                            </ul>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Notebook">Notebook</a></li>
                         </li>
+
                         <li><a href="https://2016.igem.org/Team:TJUSLS_China/Results">Results</a></li>
<li><a href="https://2016.igem.org/Team:TJUSLS_China/Modeling">MODELING</a>
+
                    </ul>
                            <ul>
+
                </li>
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Modeling">Modeling</a></li>
+
                <li onmouseover="getHover(2)" class=""><a href="https://2016.igem.org/Team:TJUSLS_China/Parts">PARTS</a>
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Software">Software</a></li>
+
                    <ul>
                             
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Basic_Part">Basic Parts</a></li>
 +
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Composite_Part">Composite Parts</a></li>
 +
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Part_Collection">Part Collection</a></li>
  
                            </ul>
+
                    </ul>
                        </li>
+
                </li>
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Safety">SAFETY</a></li>
+
                <li class="active" onmouseover="getHover(3)"><a class="active" href="https://2016.igem.org/Team:TJUSLS_China/Safety">SAFETY</a></li>
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Attributions">ATTRIBUTIONS</a>
+
                <li onmouseover="getHover(4)" class=""><a href="https://2016.igem.org/Team:TJUSLS_China/Attributions">ATTRIBUTIONS</a>
                        </li>
+
  
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Human_Practices">HUMAN PRACTICES</a>
+
                </li>
                            <ul>
+
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/HP/Silver">Silver</a></li>
+
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/HP/Gold">Gold</a></li>
+
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Integrated_Practices">Integrated Practices</a></li>
+
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Engagement">Engagement</a></li>
+
                             
+
  
                            </ul>
+
                <li onmouseover="getHover(5)" class=""><a href="https://2016.igem.org/Team:TJUSLS_China/Human_Practices">HUMAN PRACTICES</a>
</li>
+
                    <ul>
                          
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/HP/Silver">Silver</a></li>
                         <li><a href="https://2016.igem.org/Team:TJUSLS_China/Team">TEAM</a>
+
                         <li><a href="https://2016.igem.org/Team:TJUSLS_China/HP/Gold">Gold</a></li>
 +
                         <li><a href="https://2016.igem.org/Team:TJUSLS_China/Integrated_Practices">Integrated Practices</a></li>
 +
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Engagement">Engagement</a></li>
 +
                    </ul>
 +
                </li>
 +
                <li onmouseover="getHover(6)" class=""><a href="https://2016.igem.org/Team:TJUSLS_China/Modeling">MODELING</a>
 +
                    <ul>
 +
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Modeling">Modeling</a></li>
 +
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Software">Software</a></li>
 +
                    </ul>
 +
                </li>
 +
                <li onmouseover="getHover(7)" class=""><a href="https://2016.igem.org/Team:TJUSLS_China/Team">TEAM</a>
  
                            <ul>
+
                    <ul>
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Team">Team</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Team">Team</a></li>
                                <li><a href="https://2016.igem.org/Team:TJUSLS_China/Collaborations">Collaborations</a></li>
+
                        <li><a href="https://2016.igem.org/Team:TJUSLS_China/Collaborations">Collaborations</a></li>
                            </ul>
+
                        </li>
+
 
                     </ul>
 
                     </ul>
                    <script type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_nav?action=raw&ctype=text/javascript"></script>
+
                </li>
                </div>
+
            </ul>
                <!-- end h_menu4 -->
+
            <!--<script type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/home/home_nav?action=raw&amp;ctype=text/javascript"></script>-->
 
+
                <div class="clear"></div>
+
            </div>
+
 
         </div>
 
         </div>
 +
        <!-- end h_menu4 -->
 +
        <div class="clear"></div>
 
     </div>
 
     </div>
 +
    <div class="sec-menu">
 +
        <img class="sec-menu-dot" src="https://static.igem.org/mediawiki/2016/f/f9/T--TJUSLS_China--sec-menu.png"/>
 +
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        <ul>
 +
            <li><a href="#part1">Project Design</a></li>
 +
            <li><a href="#part2">Lab Work</a></li>
 +
            <li><a href="#part3">Shipment</a></li>
 +
        </ul>
 +
    </div>
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</div>
 +
<!--title-->
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<div class="second-title">
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    Results
 +
    <div class="second-title-img">
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<div class="second-content">
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    <div class="sec-list-wenzi">
 +
        <h3 id="part1">Mutation:</h3>
 +
        <img src="https://static.igem.org/mediawiki/igem.org/2/22/ProofTJU1.jpg" width="600" height="300">
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        <div class="sec-wenzi-content">Figure 1. The Comparison of the enzyme activity between PETase and three kinds of mutated PETase. The reaction condition is 100μL solution,pH 9.0(bicine-NaOH), 40 degree, 18h, the substrate is a round with a diameter of 2mm. The results are detected by Hplc. The y-axis stands for the area of the peak of MHET, the main product of the PETase’s degrading of PET. The x-axis stands for the concentration of the protein.</div>
  
    <!--header-banner-->
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        <img src="https://static.igem.org/mediawiki/igem.org/5/53/ProofTJU2.jpg" height="300" width="600">
    <div class="header-banner">
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         <div class="sec-wenzi-content">Figure 2. PETase’s self-degrading condition in different temperature. We take the quantity of PETase in the day the experiment begines as 100%. We can now see when stored in low temperature, the protein was degraded slowly, but in room temperature, the protein degrades rapidly. The Quantity of PETase left was measured by protein gel and analysised by a computer program called GEL-PRO.</div>
         <div class="banner-grids">
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            <div class="baneer-grid1">
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            </div>
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        <div class="sec-wenzi-list">Surface display in E.coli by using INP</div>
            <div class="baneer-grid2">
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        <img src="https://static.igem.org/mediawiki/igem.org/6/64/ProofTJU3.jpg" height="300" width="600">
 +
        <div class="sec-wenzi-content">Figure 3  Relative enzyme activity of  engineering bacteria E.coli(BL21)/pET22b(+)NP when induced at 16℃.</div>
  
            </div>
+
        <img src="https://static.igem.org/mediawiki/igem.org/3/3c/ProofTJU4.jpg" height="300" width="600">
            <div class="baneer-grid3">
+
        <div class="sec-wenzi-content">Figure 4  Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)NP when induced at 25℃ with different amount of bacteria.</div>
  
            </div>
+
        <img src="https://static.igem.org/mediawiki/igem.org/e/e0/ProofTJU5.jpg" height="300" width="600">
 +
        <div class="sec-wenzi-content">Figure 5  Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)NP when induced with 0.1mM IPTG for 24h.</div>
  
            <div class="clearfix"></div>
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         <img src="https://static.igem.org/mediawiki/igem.org/8/82/ProofTJU6.jpg" height="300" width="600">
         </div>
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        <div class="sec-wenzi-content">Figure 6  Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)NP when induced at 16℃ with 0.1mM IPTG for 1h, 4h, 8h, 12h, 16h and 20h.</div>
        <div class="top-header">
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                        <h3>Project</h3>
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                        <a href="https://2016.igem.org/wiki/index.php?title=Team:TJUSLS_China" class="active">home</a>
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                        <a href="#part1">part1</a>
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                        <a href="#part2">part2</a>
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                        <a href="#part3">part3</a>
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                        <a href="#part4">part4</a>
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                        <!-- Class "cbp-spmenu-open" gets applied to menu and "cbp-spmenu-push-toleft" or "cbp-spmenu-push-toright" to the body -->
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                    <!-- Classie - class helper functions by @desandro https://github.com/desandro/classie -->
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         <div class="sec-wenzi-list">Surface display in E.coli by using LPP-OmpA</div>
                    <script language="javascript" type="text/javascript" src="https://2016.igem.org/Team:TJUSLS_China/js/classie?
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action=raw&ctype=text/javascript"></script>
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        <div class="sec-wenzi-content">Figure 7  Relative enzyme activity of  engineering bacteria E.coli(BL21)/pET22b(+)LAP when induced at 16℃.</div>
                    <!-- /script-for-menu -->
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                </div>
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                <div class="clearfix"> </div>
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         </div>
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+
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+
    <div class="content">
+
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        <!--<div class="welcome">
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            <div class="container">
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                    </div>
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                    <div class="col-md-6 welcom-grid1">
+
                        <h3>welcome!</h3>
+
                        <p>Quis autem vel eum iure reprehenderit, qui in ea voluptate velit esse, quam nihil molestiae consequatur, vel illum, qui dolorem eum fugiat, quo voluptas nulla pariatur. Sed in lacus ut enim adipiscing aliquet. Nulla venena tis. In pede mi, aliquet sit amet, euismod in, auctor ut, ligula. Aliquam dapibus tincidunt metus. Praesent justo dolor, lobortis quis, lobortis dignissim, pulvinar ac, lorem. Vestibulum sed ante. Donec sagittis euismod purus.Dolor nunc vule putateulr ips dol consec.Donec semp ertet laciniate ultricie upien disse comete dolo lectus fgilla itollicil tua ludin dolor nec met quam accumsan dolore. </p>
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                    </div>
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                </div>
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            </div>
+
  
         </div>-->
+
         <div class="sec-wenzi-list">Surface display in E.coli by using BrkA</div>
        <!--end-welcome-->
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        <img src="https://static.igem.org/mediawiki/igem.org/6/60/ProofTJU9.jpg" height="300" width="600">
        <!--charities-->
+
        <div class="sec-wenzi-content">Figure 8 Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)Brk when induced at 16℃ and 25 ℃ with 0.02mM IPTG.And the last two were induced with 0.09mM IPTG.</div>
        <!--<div class="charities">
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            <div class="container">
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                <h3>our charities</h3>
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                        <h4>Adoption Child</h4>
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                        <p>Raemuis ultricaretra gc accumsan malda or sit amet errepsm do elentu vel curslor sitmt conaiing eli.</p>
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                        <h4>Charity actions</h4>
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                        <h4>Child Protection</h4>
+
                        <p>Raemuis ultricaretra gc accumsan malda or sit amet errepsm do elentu vel curslor sitmt conaiing eli.</p>
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                        <h4>Pellentesque sed</h4>
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                        <p>Raemuis ultricaretra gc accumsan malda or sit amet errepsm do elentu vel curslor sitmt conaiing eli.</p>
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                    <div class="clearfix"></div>
+
                </div>
+
            </div>
+
        </div>-->
+
        <!--charities-->
+
        <!--<div class="Testimonials" id="testimonials">
+
            <div class="container">
+
                <div class="testimonial-header">
+
                    <h3>testimonial</h3>
+
                </div>
+
                <div class="test-monials">
+
                    &lt;!&ndash; start content_slider &ndash;&gt;
+
                    <div id="owl-demo" class="owl-carousel">
+
                        <div class="item">
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+
                                <h4><a href="#">John Doe</a></h4>
+
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+
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+
                        <div class="item">
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                            <div class="testmonial-text">
+
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+
                                <h4><a href="#">Mark Trand</a></h4>
+
                            </div>
+
                            <div class="clearfix"> </div>
+
                        </div>
+
                        <div class="item">
+
                            <div class="testmonial-text">
+
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+
                                <h4><a href="#">John Doe</a></h4>
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                            </div>
+
                            <div class="clearfix"> </div>
+
                        </div>
+
  
                    </div>
+
        <div class="sec-wenzi-list">Surface display in E.coli by using AIDA</div>
 +
        <img src="https://static.igem.org/mediawiki/igem.org/d/d5/ProofTJU10.jpg" height="300" width="600">
 +
        <div class="sec-wenzi-content">Figure 9. Reletive enzyme activity of engineering  bacteria E.coli(BL21)/pET22b(+) ap at 16 ℃</div>
  
                    &lt;!&ndash;&#45;&#45;sreen-gallery-cursual&#45;&#45;&ndash;&gt;
+
        <div class="sec-wenzi-list">Surface display in Pichia Pastoris</div>
                    <div class="sreen-gallery-cursual">
+
         <img src="https://static.igem.org/mediawiki/igem.org/0/0f/ProofTJU11.jpg" height="300" width="600">
                        &lt;!&ndash; requried-jsfiles-for owl &ndash;&gt;
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                        <link rel="stylesheet" href="https://2016.igem.org/Team:TJUSLS_China/CSS_Home_owl_carousel?action=raw&amp;ctype=text/css">
+
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+
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+
                            $(document).ready(function() {
+
                                $("#owl-demo").owlCarousel({
+
                                    items : 1,
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+
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+
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+
                        &lt;!&ndash; //requried-jsfiles-for owl &ndash;&gt;
+
                    </div>
+
                </div>
+
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+
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+
        <!---- testmonials ---->
+
        <div class="copyrights">Collect from <a href="http://www.cssmoban.com/" >aaaa</a></div>
+
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+
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                <h3>Recent from the event</h3>
+
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+
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+
                        <p class="link">09/05/2015 by<a href="#">Admin</a>13 comments</p>
+
                        <h4><a href="#">Vestibulum iaculis</a></h4>
+
                        <p>Raemuis ultricaretra gc accumsan malda or sit amet errepsm do elentu vel curslor sitmt conaiing.</p>
+
                    </div>
+
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+
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+
                        <p class="link">19/8/2013 by<a href="#">Admin</a>10 comments</p>
+
                        <h4><a href="#">Pellentesque sed dolor</a></h4>
+
                        <p>Raemuis ultricaretra gc accumsan malda or sit amet errepsm do elentu vel curslor sitmt conaiing.</p>
+
                    </div>
+
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+
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+
                        <p class="link">29/9/2015 by<a href="#">Admin</a>09 comments</p>
+
                        <h4><a hrf="#">Vestibulum iaculis</a></h4>
+
                        <p>Raemuis ultricaretra gc accumsan malda or sit amet errepsm do elentu vel curslor sitmt conaiing.</p>
+
                    </div>
+
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+
                    <div style="background-color: #eee">
+
                        <div id="part1">
+
                            <p>
+
                                1111111111111111111111111111111111111111111111111
+
                                PETase
+
                                PET降解酶发现于一种以PET为主要碳源的微生物(Ideonella sakaiensis 201-F6),该酶能将大分子聚合物降解为单体。表面展示技术是通过目的蛋白或多肽的编码基因与锚定蛋白的基因融合,通过融合蛋白的后翻译、折叠,将目的蛋白展示在宿主细胞壁的表面,从而获得全细胞催化剂的一种方法。我团队课题以PETase为主体,利用表面展示技术,从三个方面进行研究:
+
                                第一,通过蛋白质晶体学以及X射线衍射技术来解析PETase的结构,找到酶的催化中心和结合中心,并依据其结构特性指导突变位点的选择,从而对PETase进行定向突变以提高其降解效率和热稳定性;第二,将其进行原核(大肠杆菌)和真核(毕赤酵母)表面展示以进行全细胞酶催化反应。第三,将其与疏水蛋白在毕赤酵母中进行融合分泌,利用疏水蛋白两面性中的强疏水性为PET降解反应提供疏水环境以促进降解效率。同时,将疏水蛋白与PETase在毕赤酵母中进行共展示,利用疏水蛋白能改造细胞表面性质以适应极端环境的特点进一步提高全细胞催化剂的降解效率,打破了反应条件的限制,极大地拓宽了PETase降解反应的条件,以实现PETase的工业应用。
+
                            </p>
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                            <img src="https://static.igem.org/mediawiki/2016/7/77/B1.jpg" alt="/">
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                        </div>
+
  
                        <div id="part2">
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        <img src="https://static.igem.org/mediawiki/igem.org/e/e5/ProofTJU12.jpg" height="300" width="600">
                            <p>
+
        <div class="sec-wenzi-content">Figure 10: The activity of P. pastoris PETase-GCW21. a&b used the first group of yeast; c&d used the second of yeast; a&c:the activity in different yeasts'concentration under the best hour; b&d: the activity in different hours under the best concentration.</div>
                                2222222222222222222222222222222222222222222222222
+
 
                                PETase
+
 
                                PET降解酶发现于一种以PET为主要碳源的微生物(Ideonella sakaiensis 201-F6),该酶能将大分子聚合物降解为单体。表面展示技术是通过目的蛋白或多肽的编码基因与锚定蛋白的基因融合,通过融合蛋白的后翻译、折叠,将目的蛋白展示在宿主细胞壁的表面,从而获得全细胞催化剂的一种方法。我团队课题以PETase为主体,利用表面展示技术,从三个方面进行研究:
+
        <img src="https://static.igem.org/mediawiki/igem.org/4/48/ProofTJU13.jpg" height="300" width="600">
                                第一,通过蛋白质晶体学以及X射线衍射技术来解析PETase的结构,找到酶的催化中心和结合中心,并依据其结构特性指导突变位点的选择,从而对PETase进行定向突变以提高其降解效率和热稳定性;第二,将其进行原核(大肠杆菌)和真核(毕赤酵母)表面展示以进行全细胞酶催化反应。第三,将其与疏水蛋白在毕赤酵母中进行融合分泌,利用疏水蛋白两面性中的强疏水性为PET降解反应提供疏水环境以促进降解效率。同时,将疏水蛋白与PETase在毕赤酵母中进行共展示,利用疏水蛋白能改造细胞表面性质以适应极端环境的特点进一步提高全细胞催化剂的降解效率,打破了反应条件的限制,极大地拓宽了PETase降解反应的条件,以实现PETase的工业应用。
+
        <img src="https://static.igem.org/mediawiki/igem.org/2/2d/ProofTJU14.jpg" height="300" width="600">
                            </p>
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                            <img src="https://static.igem.org/mediawiki/2016/7/77/B1.jpg" alt="/">
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        <div class="sec-wenzi-content">Figure 11: The activity of P. pastoris PETase-GCW51. a&b used the first group of yeast; c&d used the third of yeast; a&c:the activity in different yeasts'concentration under the best hour; b&d: the activity in different hours under the best concentration.</div>
                        </div>
+
 
                        <div id="part3">
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        <img src="https://static.igem.org/mediawiki/igem.org/d/d7/ProofTJU15.jpg" height="300" width="600">
                            <p>
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        <img src="https://static.igem.org/mediawiki/igem.org/0/0c/ProofTJU16.jpg" height="300" width="600">
                                333333333333333333333333333333333333333333333333
+
        <div class="sec-wenzi-content">Figure 12: The activity of P. pastoris PETase-GCW61. a&b used the first group of yeast; c&d used the third of yeast; a&c:the activity in different yeasts'concentration under the best hour; b&d: the activity in different hours under the best concentration.</div>
                                PETase
+
 
                                PET降解酶发现于一种以PET为主要碳源的微生物(Ideonella sakaiensis 201-F6),该酶能将大分子聚合物降解为单体。表面展示技术是通过目的蛋白或多肽的编码基因与锚定蛋白的基因融合,通过融合蛋白的后翻译、折叠,将目的蛋白展示在宿主细胞壁的表面,从而获得全细胞催化剂的一种方法。我团队课题以PETase为主体,利用表面展示技术,从三个方面进行研究:
+
         <div class="sec-wenzi-list">Co-display in Pichia Pastoris</div>
                                第一,通过蛋白质晶体学以及X射线衍射技术来解析PETase的结构,找到酶的催化中心和结合中心,并依据其结构特性指导突变位点的选择,从而对PETase进行定向突变以提高其降解效率和热稳定性;第二,将其进行原核(大肠杆菌)和真核(毕赤酵母)表面展示以进行全细胞酶催化反应。第三,将其与疏水蛋白在毕赤酵母中进行融合分泌,利用疏水蛋白两面性中的强疏水性为PET降解反应提供疏水环境以促进降解效率。同时,将疏水蛋白与PETase在毕赤酵母中进行共展示,利用疏水蛋白能改造细胞表面性质以适应极端环境的特点进一步提高全细胞催化剂的降解效率,打破了反应条件的限制,极大地拓宽了PETase降解反应的条件,以实现PETase的工业应用。
+
         <img src="https://static.igem.org/mediawiki/igem.org/6/67/ProofTJU17.jpg" height="300" width="600">
                            </p>
+
        <div class="sec-wenzi-content">Figure 13: The activity of the first group of ppic9-PETase-GCW51 & ppiczaA-HFB1-GCW61 co-display transformants in different hours  and amount of yeast.</div>
                            <img src="https://static.igem.org/mediawiki/2016/7/77/B1.jpg" alt="/">
+
                        </div>
+
                        <div id="part4">
+
                            <p>
+
                                44444444444444444444444444444444444444444444444444
+
                                PETase
+
                                PET降解酶发现于一种以PET为主要碳源的微生物(Ideonella sakaiensis 201-F6),该酶能将大分子聚合物降解为单体。表面展示技术是通过目的蛋白或多肽的编码基因与锚定蛋白的基因融合,通过融合蛋白的后翻译、折叠,将目的蛋白展示在宿主细胞壁的表面,从而获得全细胞催化剂的一种方法。我团队课题以PETase为主体,利用表面展示技术,从三个方面进行研究:
+
                                第一,通过蛋白质晶体学以及X射线衍射技术来解析PETase的结构,找到酶的催化中心和结合中心,并依据其结构特性指导突变位点的选择,从而对PETase进行定向突变以提高其降解效率和热稳定性;第二,将其进行原核(大肠杆菌)和真核(毕赤酵母)表面展示以进行全细胞酶催化反应。第三,将其与疏水蛋白在毕赤酵母中进行融合分泌,利用疏水蛋白两面性中的强疏水性为PET降解反应提供疏水环境以促进降解效率。同时,将疏水蛋白与PETase在毕赤酵母中进行共展示,利用疏水蛋白能改造细胞表面性质以适应极端环境的特点进一步提高全细胞催化剂的降解效率,打破了反应条件的限制,极大地拓宽了PETase降解反应的条件,以实现PETase的工业应用。
+
                            </p>
+
                            <img src="https://static.igem.org/mediawiki/2016/7/77/B1.jpg" alt="/">
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                        </div>
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                        <div id="part5">
+
                            <p>
+
                                55555555555555555555555555555555555555555555555555
+
                                PETase
+
                                PET降解酶发现于一种以PET为主要碳源的微生物(Ideonella sakaiensis 201-F6),该酶能将大分子聚合物降解为单体。表面展示技术是通过目的蛋白或多肽的编码基因与锚定蛋白的基因融合,通过融合蛋白的后翻译、折叠,将目的蛋白展示在宿主细胞壁的表面,从而获得全细胞催化剂的一种方法。我团队课题以PETase为主体,利用表面展示技术,从三个方面进行研究:
+
                                第一,通过蛋白质晶体学以及X射线衍射技术来解析PETase的结构,找到酶的催化中心和结合中心,并依据其结构特性指导突变位点的选择,从而对PETase进行定向突变以提高其降解效率和热稳定性;第二,将其进行原核(大肠杆菌)和真核(毕赤酵母)表面展示以进行全细胞酶催化反应。第三,将其与疏水蛋白在毕赤酵母中进行融合分泌,利用疏水蛋白两面性中的强疏水性为PET降解反应提供疏水环境以促进降解效率。同时,将疏水蛋白与PETase在毕赤酵母中进行共展示,利用疏水蛋白能改造细胞表面性质以适应极端环境的特点进一步提高全细胞催化剂的降解效率,打破了反应条件的限制,极大地拓宽了PETase降解反应的条件,以实现PETase的工业应用。
+
                            </p>
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        <div class="sec-wenzi-content">Figure 14: The activity of the second group of ppic9-PETase-GCW51 & ppiczaA-HFB1-GCW61 co-display transformants in different hours  and amount of yeast.</div>
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        <div class="sec-wenzi-content">Figure 15: The activity of the first group of ppic9-PETase-GCW51 & ppiczaA-HGF1-GCW61 co-display transformants in different hours  and amount of yeast.</div>
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        <div class="sec-wenzi-content">Figure 16: The activity of the second group of ppic9-PETase-GCW51 & ppiczaA-HGF1-GCW61 co-display transformants in different hours  and amount of yeast.</div>
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        <img src="https://static.igem.org/mediawiki/igem.org/e/ea/ProofTJU21.jpg" height="300" width="600">
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        <div class="sec-wenzi-content">Figure 17: The activity of ppic9-PETase-GCW51 & ppiczaA-HGF1-GCW61 co-display transformant and ppic9-PETase-GCW51 & ppiczaA-HGF1-GCW61 co-display transformant in best condition</div>
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Revision as of 14:07, 18 October 2016

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Results

Mutation:

Figure 1. The Comparison of the enzyme activity between PETase and three kinds of mutated PETase. The reaction condition is 100μL solution,pH 9.0(bicine-NaOH), 40 degree, 18h, the substrate is a round with a diameter of 2mm. The results are detected by Hplc. The y-axis stands for the area of the peak of MHET, the main product of the PETase’s degrading of PET. The x-axis stands for the concentration of the protein.
Figure 2. PETase’s self-degrading condition in different temperature. We take the quantity of PETase in the day the experiment begines as 100%. We can now see when stored in low temperature, the protein was degraded slowly, but in room temperature, the protein degrades rapidly. The Quantity of PETase left was measured by protein gel and analysised by a computer program called GEL-PRO.
Surface display in E.coli by using INP
Figure 3 Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)NP when induced at 16℃.
Figure 4 Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)NP when induced at 25℃ with different amount of bacteria.
Figure 5 Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)NP when induced with 0.1mM IPTG for 24h.
Figure 6 Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)NP when induced at 16℃ with 0.1mM IPTG for 1h, 4h, 8h, 12h, 16h and 20h.
Surface display in E.coli by using LPP-OmpA
Figure 7 Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)LAP when induced at 16℃.
Surface display in E.coli by using BrkA
Figure 8 Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)Brk when induced at 16℃ and 25 ℃ with 0.02mM IPTG.And the last two were induced with 0.09mM IPTG.
Surface display in E.coli by using AIDA
Figure 9. Reletive enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+) ap at 16 ℃
Surface display in Pichia Pastoris
Figure 10: The activity of P. pastoris PETase-GCW21. a&b used the first group of yeast; c&d used the second of yeast; a&c:the activity in different yeasts'concentration under the best hour; b&d: the activity in different hours under the best concentration.
Figure 11: The activity of P. pastoris PETase-GCW51. a&b used the first group of yeast; c&d used the third of yeast; a&c:the activity in different yeasts'concentration under the best hour; b&d: the activity in different hours under the best concentration.
Figure 12: The activity of P. pastoris PETase-GCW61. a&b used the first group of yeast; c&d used the third of yeast; a&c:the activity in different yeasts'concentration under the best hour; b&d: the activity in different hours under the best concentration.
Co-display in Pichia Pastoris
Figure 13: The activity of the first group of ppic9-PETase-GCW51 & ppiczaA-HFB1-GCW61 co-display transformants in different hours and amount of yeast.
Figure 14: The activity of the second group of ppic9-PETase-GCW51 & ppiczaA-HFB1-GCW61 co-display transformants in different hours and amount of yeast.
Figure 15: The activity of the first group of ppic9-PETase-GCW51 & ppiczaA-HGF1-GCW61 co-display transformants in different hours and amount of yeast.
Figure 16: The activity of the second group of ppic9-PETase-GCW51 & ppiczaA-HGF1-GCW61 co-display transformants in different hours and amount of yeast.
Figure 17: The activity of ppic9-PETase-GCW51 & ppiczaA-HGF1-GCW61 co-display transformant and ppic9-PETase-GCW51 & ppiczaA-HGF1-GCW61 co-display transformant in best condition