LeeKartoon (Talk | contribs) |
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<html> | <html> | ||
− | + | ||
<style type="text/css"> | <style type="text/css"> | ||
#top_title{ | #top_title{ | ||
display:none; | display:none; | ||
} | } | ||
− | + | * { | |
− | + | box-sizing: border-box; | |
− | + | } | |
− | + | a { | |
− | + | text-decoration: none; | |
− | + | } | |
− | + | ul, | |
− | + | ol { | |
− | + | list-style: none; | |
− | + | } | |
− | + | body { | |
− | + | font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; | |
− | + | font-size: 14px; | |
− | + | line-height: 1.42857143; | |
− | + | color: #333; | |
− | + | min-width: 768px; | |
− | + | } | |
− | + | .navbar-default { | |
− | + | background-color: #EEE; | |
− | + | border-color: #e7e7e7; | |
− | + | top: 16px; | |
− | + | border-width: 0 0 1px; | |
− | + | position: fixed; | |
− | + | right: 0; | |
− | + | left: 0; | |
− | + | z-index: 1030; | |
− | + | min-height: 50px; | |
− | + | margin-bottom: 20px; | |
− | + | border: 1px solid transparent; | |
− | + | min-width: 768px; | |
− | + | } | |
− | + | .container { | |
− | + | padding-right: 15px; | |
− | + | padding-left: 15px; | |
− | + | margin-right: auto; | |
− | + | margin-left: auto; | |
− | + | } | |
− | + | @media screen and (min-width: 768px) { | |
− | + | .container { | |
− | + | width: 750px; | |
− | + | } | |
− | + | } | |
− | + | @media screen and (min-width: 992px) { | |
− | + | .container { | |
− | + | width: 970px; | |
− | + | } | |
− | + | } | |
− | + | @media screen and (min-width: 1200px) { | |
− | + | .container { | |
− | + | width: 1170px; | |
− | + | } | |
− | + | } | |
− | + | .navbar-header { | |
− | + | float: left; | |
− | + | } | |
− | + | .navbar-brand { | |
− | + | color: #777; | |
− | + | float: left; | |
− | + | height: 50px; | |
− | + | padding: 15px 15px; | |
− | + | font-size: 18px; | |
− | + | line-height: 20px; | |
− | + | } | |
− | + | .navbar>ul>li { | |
− | + | float: left; | |
− | + | } | |
− | + | .nav>li { | |
− | + | position: relative; | |
− | + | display: block; | |
− | + | margin-right: 10px; | |
− | + | } | |
− | + | .nav>li>a { | |
− | + | color: #777; | |
− | + | position: relative; | |
− | + | display: block; | |
− | + | padding: 8px 15px; | |
− | + | cursor: pointer; | |
− | + | } | |
− | + | .nav>li>a:hover { | |
− | + | color: #333; | |
− | + | } | |
− | + | .caret { | |
− | + | display: inline-block; | |
− | + | width: 0; | |
− | + | height: 0; | |
− | + | margin-left: 2px; | |
− | + | vertical-align: middle; | |
− | + | border-top: 4px solid #777; | |
− | + | border-right: 4px solid transparent; | |
− | + | border-left: 4px solid transparent; | |
− | + | position: absolute; | |
− | + | top: 18px; | |
− | + | right: 1px; | |
− | + | } | |
− | + | .nav>li>a:hover+span { | |
− | + | border-top: 4px solid #333; | |
− | + | } | |
− | + | .nav>li .dropdown-menu { | |
− | + | margin-top: 0; | |
− | + | border-top-left-radius: 0; | |
− | + | border-top-right-radius: 0; | |
− | + | position: absolute; | |
− | + | top: 35px; | |
− | + | left: 0; | |
− | + | z-index: 1000; | |
− | + | float: left; | |
− | + | min-width: 160px; | |
− | + | padding: 5px 0; | |
− | + | margin: 2px 0 0; | |
− | + | font-size: 14px; | |
− | + | text-align: left; | |
− | + | list-style: none; | |
− | + | background-color: #fff; | |
− | + | -webkit-background-clip: padding-box; | |
− | + | background-clip: padding-box; | |
− | + | border: 1px solid #ccc; | |
− | + | border: 1px solid rgba(0, 0, 0, .15); | |
− | + | border-radius: 4px; | |
− | + | -webkit-box-shadow: 0 6px 12px rgba(0, 0, 0, .175); | |
− | + | box-shadow: 0 6px 12px rgba(0, 0, 0, .175); | |
− | + | display: none; | |
− | + | } | |
− | + | .dropdown-menu>li { | |
− | + | display: list-item; | |
− | + | } | |
− | + | .dropdown-menu>li>a { | |
− | + | display: block; | |
− | + | padding: 3px 20px; | |
− | + | clear: both; | |
− | + | font-weight: normal; | |
− | + | line-height: 1.42857143; | |
− | + | color: #333; | |
− | + | white-space: nowrap; | |
− | + | } | |
− | + | .dropdown-menu>li>a:hover { | |
− | + | background-color: #eee; | |
− | + | } | |
html { | html { | ||
− | + | font-family: sans-serif; | |
− | + | -webkit-text-size-adjust: 100%; | |
− | + | -ms-text-size-adjust: 100%; | |
} | } | ||
#mainDiv{ | #mainDiv{ | ||
− | + | ||
− | + | margin: 0; | |
− | + | height: 2500px; | |
− | + | padding-top: 70px; | |
− | + | min-width: 1140px; | |
− | + | background: url('https://static.igem.org/mediawiki/2018/4/4b/T--SCAU-China--ziyeMain.jpg') center center; | |
} | } | ||
.heart { | .heart { | ||
− | + | width: 1140px; | |
− | + | margin: 0 auto;/* 通用的版心 */ | |
} | } | ||
#mask { | #mask { | ||
− | + | width: 100%; | |
− | + | height: 2450px; | |
− | + | background: rgba(0, 0, 0, 0.3); | |
− | + | margin-top: -15px; | |
− | + | padding-top: 15px; | |
} | } | ||
.DBoard {/* 展板样式 */ | .DBoard {/* 展板样式 */ | ||
− | + | margin: 0px auto 40px auto; | |
− | + | width: 1000px; | |
− | + | height: 500px; | |
− | + | border-radius: 3px; | |
− | + | background-color: #fff; | |
− | + | overflow: hidden; | |
− | + | position: relative; | |
} | } | ||
.DBoard0 {/*文字样式*/ | .DBoard0 {/*文字样式*/ | ||
− | + | margin: 0px auto 40px auto; | |
− | + | width: 1000px; | |
− | + | height: 500px; | |
− | + | border-radius: 3px; | |
− | + | color: white; | |
− | + | overflow: hidden; | |
− | + | position: relative; | |
} | } | ||
#mask p { | #mask p { | ||
− | + | color: #fff; | |
− | + | text-align: center; | |
} | } | ||
#lists-DB { | #lists-DB { | ||
− | + | cursor: pointer; | |
} | } | ||
#lists-DB:hover #arr-l,#lists-DB:hover #arr-r{ | #lists-DB:hover #arr-l,#lists-DB:hover #arr-r{ | ||
− | + | display: block; | |
} | } | ||
#lists { | #lists { | ||
− | + | position: relative; | |
− | + | width: 4000px; | |
} | } | ||
#lists img { | #lists img { | ||
− | + | float: left; | |
} | } | ||
#arr-l,#arr-r { | #arr-l,#arr-r { | ||
− | + | display: block; | |
− | + | position: absolute; | |
− | + | width: 40px; | |
− | + | height: 60px; | |
− | + | background: rgba(0, 0, 0, .4); | |
− | + | top: 50%; | |
− | + | margin-top: -30px; | |
− | + | color: #fff; | |
− | + | font: 600 30px/60px "宋体"; | |
− | + | line-height: 60px; | |
− | + | text-align: center; | |
− | + | display: none; | |
} | } | ||
#arr-l:hover,#arr-r:hover { | #arr-l:hover,#arr-r:hover { | ||
− | + | background: rgba(0, 0, 0, .7); | |
} | } | ||
#arr-l { | #arr-l { | ||
− | + | left: 0; | |
} | } | ||
#arr-r { | #arr-r { | ||
− | + | right: 0; | |
} | } | ||
#buttons { | #buttons { | ||
− | + | position: absolute; | |
− | + | bottom: 15px; | |
− | + | left: 50%; | |
} | } | ||
#buttons span { | #buttons span { | ||
− | + | width: 12px; | |
− | + | height: 12px; | |
− | + | background: #ccc; | |
− | + | float: left; | |
− | + | border-radius: 50%; | |
− | + | margin: 0 5px 0 5px; | |
} | } | ||
.on { | .on { | ||
− | + | background-color: #333 !important; | |
} | } | ||
#toTop { | #toTop { | ||
− | + | width: 50px; | |
− | + | height: 50px; | |
− | + | background: url("https://static.igem.org/mediawiki/2018/b/b6/T--SCAU-China--toTop.png") no-repeat center center rgba(0, 0, 0, .3); | |
− | + | position: fixed; | |
− | + | right: 50px; | |
− | + | bottom: 5px; | |
− | + | cursor: pointer; | |
− | + | display: none; | |
} | } | ||
− | + | ||
} | } | ||
#img4{ | #img4{ | ||
− | + | width: 10%; | |
} | } | ||
#content { | #content { | ||
− | + | width: 101%; | |
− | + | padding: 15px; | |
− | + | border: none; | |
− | + | color: black; | |
− | + | margin-left: 20px; | |
− | + | margin-right: auto; | |
− | + | background-color: #ffffff; | |
− | + | position: relative; | |
} | } | ||
#content { | #content { | ||
− | + | position: relative; | |
− | + | width: 101%; | |
− | + | margin: 0 auto; | |
− | + | padding: 5px 5px 5px 5px; | |
− | + | background: white; | |
− | + | color: black; | |
− | + | border-left: 1px solid #444444; | |
− | + | border-right: 1px solid #444444; | |
− | + | line-height: 1.5em; | |
− | + | z-index: 2; | |
} | } | ||
#title{ | #title{ | ||
− | + | height: 100px; | |
− | + | background: rgba(0, 0, 0, 0.3); | |
− | + | text-align: left; | |
− | + | font-size: 60px; | |
− | + | color: white; | |
− | + | font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; | |
− | + | line-height: 100px; | |
} | } | ||
#article1{ | #article1{ | ||
− | + | background: rgba(0, 0, 0, 0); | |
height: 1710px; | height: 1710px; | ||
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#article1 p{ | #article1 p{ | ||
− | + | text-align: left; | |
− | + | position: relative; | |
− | + | color: white; | |
− | + | margin-top: 20px; | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; | |
− | + | font-size:20px; | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
} | } | ||
+ | |||
+ | |||
#title2{ | #title2{ | ||
− | + | height: 50px; | |
− | + | background: rgba(0, 0, 0, 0.3); | |
− | + | text-align: left; | |
− | + | font-size:30px; | |
− | + | color: white; | |
− | + | font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; | |
− | + | line-height: 50px; | |
} | } | ||
#art1{ | #art1{ | ||
− | + | background: rgba(0, 0, 0, 0); | |
− | + | height: 210px; | |
} | } | ||
#art2{ | #art2{ | ||
− | + | background: rgba(0, 0, 0, 0); | |
− | + | height: 580px; | |
+ | } | ||
+ | |||
+ | #art3{ | ||
+ | background: rgba(0, 0, 0, 0); | ||
+ | height: 3000px; | ||
} | } | ||
#art1 p{ | #art1 p{ | ||
− | + | text-align: left; | |
− | + | position: relative; | |
− | + | color: white; | |
− | + | margin-top: 0px; | |
− | + | ||
− | + | font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; | |
− | + | font-size:20px; | |
} | } | ||
#art2 p{ | #art2 p{ | ||
− | + | text-align: left; | |
− | + | position: relative; | |
− | + | color: white; | |
− | + | margin-top:0px; | |
− | + | ||
− | + | font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; | |
− | + | font-size:20px; | |
} | } | ||
+ | #art3 p{ | ||
+ | text-align: left; | ||
+ | position: relative; | ||
+ | color: white; | ||
+ | margin-top:0px; | ||
+ | |||
+ | font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; | ||
+ | font-size:20px; | ||
+ | } | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
body { | body { | ||
− | + | background-color: white; | |
} | } | ||
</style> | </style> | ||
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</div> | </div> | ||
</div> | </div> | ||
+ | |||
<div id="mainDiv"> | <div id="mainDiv"> | ||
<div class="heart"><!-- 版心 --> | <div class="heart"><!-- 版心 --> | ||
− | + | <div id="mask"><!-- 半透明底板 --> | |
− | + | ||
− | + | <div class="DBoard" id="title"> | |
− | + | ||
− | + | ||
+ | Improve | ||
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</div> | </div> | ||
− | + | <div class="DBoard" id="title2"> | |
− | + | ||
− | + | (4) BBa_K2559005 /BBa_K2559009 | |
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− | + | ||
+ | </div> | ||
+ | <div class="DBoard" id="art3" height=1500px> | ||
+ | <p> | ||
− | + | The part BBa_K2559006 has a sequence improvement on the basic part submitted by iGEM07_Peking (BBa_l714891) which encodes the SDY_eGFP. However, we found out a 16 bp nucleotides redundancy in the eGFP starting coding region in BBa_I714891, after checking the sequence of BBa_I714891 from NCBI. Therefore, we decided to delete the redundant 16 bp nucleotides in BBa_I714891 to amend the length of eGFP coding sequence. The amended eGFP coding biobrick is the BBa_K2559005. | |
− | + | </p> | |
− | + | <p>To test the function of BBa_K2559005, we designed a new E.coli expression vector containing our new part termed as BBa_K2559003under a strong E.coli endogenous promoter (PrplJ). Therefore, the amended eGFP in BBa_K2559005 was driven by PrplJ promoter, and expressed in DH10B. In addition, we also applied the BBa_K2559005 in the promoter intensity analysis of our other two new parts, the BBa_K2559004 and BBa_K2559011 which are relatively weaker E.coli endogenous promoters (PdapA and PcaiF) (Figure 1).</p> | |
− | + | <img src="https://static.igem.org/mediawiki/2018/8/80/T--SCAU-China--Measurement014.png" alt="" /> | |
− | + | <br><br> | |
− | + | <img src="https://static.igem.org/mediawiki/2018/3/3f/T--SCAU-China--Measurement015.png" alt="" /> | |
− | + | <p style="font-size:16px; text-align: center;">Figure 1: Fluorescent intensity of amended eGFP driven by by PrplJ, PdapA, PcaiF promoter.</p> | |
− | + | <p>We summarized that our improvedpart, the amended eGFP coding biobrick BBa_K2559005 worked well in DH10B. We also hoped that our improvement on the BBa_I714891 can help their future applications by other groups in the future. However, it is difficult for us to perform additional experiments with BBa_K2559005 and BBa_I714891 due to the unavailable BBa_I714891.</p> | |
− | + | <p>To expand the application of BBa_K2559005, we searched theBBa_J04450stored in registry and do another improvement in the BBa_J04450. The BBa_J04450 is a strong RFP expression vector in E.coli. As the main page of BBa_J04450 mentioned, the E.coli colonies with BBa_J04450 were in red color under normal light after about 18 hour culture on LB plate (Figure 2). We used the BBa_K2559005 to replace the RFP region in BBa_J04450, the modified part is BBa_K2559009. We transferred the BBa_K2559009 to DH5α by heat-shock, and found that the fluorescence signal can be observed under the UV (Figure 2).</p> | |
− | + | <img src="https://static.igem.org/mediawiki/2018/9/94/T--SCAU-CHINA--improve3.jpg" width="700px" height="700px" /> | |
− | + | <p style="font-size:16px; text-align: center;">Figure 2 Colonies with BBa_J04450 (red colony) and BBa_K2559009 (green colony) visualized under UV lightbox.</p> | |
− | + | <p>So, we confirm that our improved part BBa_K2559005 can work in different E.coli expression system. We are also looking forward to more application of the BBa_K2559009!</p> | |
− | + | </div> | |
− | + | ||
+ | <p>School's name:SCAU</p> | ||
+ | <p>Member's name:SCAU</p> | ||
+ | <p>Designed by:SCAU</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <!-- 回到顶部按钮 --> | ||
+ | <em id="toTop"></em> | ||
− | + | <script> | |
− | + | var dropdownBoxs = document.getElementsByClassName('dropdown'); | |
− | + | var dropdownMenus = document.getElementsByClassName('dropdown-menu'); | |
− | + | for (let i = 0 ; i < dropdownBoxs.length ; i ++) { | |
+ | // console.log(dropdownBoxs[i]); | ||
+ | // console.log(dropdownMenus[i]); | ||
+ | dropdownBoxs[i].index = i; | ||
+ | dropdownBoxs[i].onclick = function () { | ||
+ | var styles = document.defaultView.getComputedStyle(dropdownMenus[i]) || dropdownMenus[i].currentStyle; | ||
+ | // console.log(styles.display); | ||
+ | if (styles.display == 'none') { | ||
+ | for (let j = 0 ; j < dropdownBoxs.length ; j ++) { | ||
+ | dropdownMenus[j].style.display = 'none'; | ||
+ | dropdownMenus[i].style.display = 'block'; | ||
+ | } | ||
+ | } else { | ||
+ | for (let j = 0 ; j < dropdownBoxs.length ; j ++) { | ||
+ | dropdownMenus[j].style.display = 'none'; | ||
+ | } | ||
+ | } | ||
+ | } | ||
+ | } | ||
+ | </script> | ||
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− | + | <script type="text/javascript"> | |
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− | + | var a= function() { | |
− | + | var box01 = document.getElementById("lists-DB"); | |
− | + | var lis = document.getElementById("lists");//获得轮播图图片盒子 | |
− | + | var btns = document.getElementById("buttons");//获得按钮盒子 | |
− | + | var imgs = lis.getElementsByTagName("img");//获得图片伪数组 | |
− | + | btns.style.width = 24 * (imgs.length - 2) + "px"; | |
− | + | btns.style.marginLeft = -(12 * (imgs.length - 2)) + "px";//动态赋值 | |
+ | for(var i = 0;i < imgs.length - 2; i++){ | ||
+ | //动态生成小圆点 | ||
+ | var span = document.createElement("span"); | ||
+ | btns.appendChild(span); | ||
+ | } | ||
− | |||
− | |||
− | + | //轮播图正式部分 | |
− | + | var prev = document.getElementById("arr-l"); | |
− | + | var next = document.getElementById("arr-r"); | |
− | + | var animated = false;//防止计时器多次被触发变量 | |
− | + | ||
− | + | ||
− | + | function animate(offset) { | |
− | + | var time = 300;//滚动一张图片总用时 | |
− | + | var inteval = 10;//滚动一次的间隔时间 | |
− | + | var speed = offset / (time / inteval);//每次滚动滑动的像素 | |
+ | var left = parseInt(lis.style.left) + offset;//先计算出滚动后的left值 | ||
+ | function go() { | ||
+ | animated = true;//为true代表正在运行 | ||
+ | //滑动函数 | ||
+ | if(((speed > 0) + (parseInt(lis.style.left) < left)===2) || ((speed < 0) + (parseInt(lis.style.left)) > left===2)){ | ||
+ | lis.style.left = parseInt(lis.style.left) + speed + "px"; | ||
+ | setTimeout(go,inteval);//设置计时器 | ||
+ | } | ||
+ | else { | ||
+ | lis.style.left = left + "px"; | ||
+ | if(parseInt(lis.style.left) > -1000) lis.style.left = -(1000 * (imgs.length - 2)) + "px"; | ||
+ | if(parseInt(lis.style.left) < -(1000 * (imgs.length - 2))) lis.style.left = -1000 + "px"; | ||
+ | animated = false;//结束运行 | ||
+ | } | ||
+ | } | ||
+ | go();//调用函数 | ||
+ | |||
+ | } | ||
+ | prev.onclick = function() { | ||
+ | if(animated) return;//正在轮播停止 | ||
+ | if(index == 0) index = spans.length - 1; | ||
+ | else index --; | ||
+ | animate(1000); | ||
+ | showButton(); | ||
+ | } | ||
+ | next.onclick = function() { | ||
+ | if(animated) return; | ||
+ | if(index == spans.length - 1) index = 0; | ||
+ | else index ++; | ||
+ | animate(-1000); | ||
+ | showButton(); | ||
+ | } | ||
+ | var index = 0;//记录现在className为on的按钮 | ||
+ | |||
+ | var spans = btns.getElementsByTagName("span");//得到btns下所有的span标签 | ||
+ | spans[0].className = "on"; | ||
+ | for(var i = 0;i < spans.length;i ++ ) { | ||
+ | spans[i].index = i;//自定义属性 | ||
+ | spans[i].onclick = function() { | ||
+ | if(animated) return; | ||
+ | if(this.className == "on") return; | ||
+ | animate((index - this.index) * 1000); | ||
+ | index = this.index; | ||
+ | showButton(); | ||
+ | } | ||
+ | } | ||
+ | |||
+ | function showButton() { //显示按钮运动的函数 | ||
+ | for(var j = 0;j < spans.length;j ++) {//排它思想 | ||
+ | //if(spans[j].className == "on") oldIndex = j; | ||
+ | spans[j].className = ""; | ||
+ | spans[index].className = "on"; | ||
+ | } | ||
+ | } | ||
+ | |||
+ | var timer;//计时器变量 , 这里为什么不能为null | ||
+ | var interval = 3000;//点击间隔时间 | ||
+ | |||
+ | function play() {//自动点击next函数 | ||
+ | timer = setTimeout(function(){ | ||
+ | next.onclick();//自动点击next | ||
+ | play();//递归调用 | ||
+ | },interval); | ||
+ | } | ||
+ | |||
+ | function stop() { | ||
+ | //停止函数 | ||
+ | clearTimeout(timer); | ||
+ | } | ||
− | + | box01.onmouseover = stop; //鼠标悬浮在轮播图上时停止 | |
− | + | box01.onmouseout = play;//鼠标离开轮播图继续 | |
− | + | play();//先自己调用一次 | |
} | } | ||
a(); | a(); | ||
− | + | </script> | |
− | + | <script type="text/javascript"> | |
− | + | //封装函数js文件,方便调用 | |
− | + | function $(id) {return document.getElementById(id);}//封装获取id对象函数 | |
− | + | function show(obj) {obj.style.display = "block";}//封装显示函数 | |
− | + | function hidden(obj) {obj.style.display = "none";}//封装隐藏函数 | |
− | + | function scroll(){ | |
− | + | if(window.pageYOffset != null) { | |
− | + | //ie9+ 和 其他浏览器 | |
− | + | return { | |
− | + | top: window.pageYOffset, | |
− | + | left: window.pageXOffset | |
− | + | } | |
− | + | } | |
− | + | else if(document.compatMode == "CSS1Compat") { | |
− | + | //非怪异浏览器:没有头部的 | |
− | + | return { | |
− | + | top: document.documentElement.scrollTop, | |
− | + | left: document.documentElement.scrollLeft | |
− | + | } | |
− | + | } | |
− | + | return {//剩下的全部都是怪异浏览器 | |
− | + | top: document.body.scrollTop, | |
− | + | left: document.body.scrollLeft | |
− | + | } | |
− | + | } | |
− | + | var leader = 0,target = 0,timer = null;//计时器变量 | |
− | + | window.onscroll = function() { | |
− | + | //滚动时执行的函数 | |
− | + | scroll().top > 0 ? show($("toTop")) : hidden($("toTop"));//隐藏和显示totop | |
− | + | leader = scroll().top; //实时记录滚动距离 | |
− | + | } | |
− | + | $("toTop").onclick = function() { | |
− | + | //toTop被点击时执行的函数 | |
− | + | target = 0; | |
− | + | timer = setInterval(function() { | |
− | + | leader = leader + (target - leader) / 10; | |
− | + | window.scrollTo(0,leader); | |
− | + | if(leader == target) clearInterval(timer);//当到达顶端清除计时器 | |
− | + | //必须写在里面,因为在计时器执行时判断 | |
− | + | },20); | |
− | + | } | |
− | + | </script> | |
</html> | </html> |
Revision as of 06:57, 17 October 2018
The part BBa_K2559006 has a sequence improvement on the basic part submitted by iGEM07_Peking (BBa_l714891) which encodes the SDY_eGFP. However, we found out a 16 bp nucleotides redundancy in the eGFP starting coding region in BBa_I714891, after checking the sequence of BBa_I714891 from NCBI. Therefore, we decided to delete the redundant 16 bp nucleotides in BBa_I714891 to amend the length of eGFP coding sequence. The amended eGFP coding biobrick is the BBa_K2559005.
To test the function of BBa_K2559005, we designed a new E.coli expression vector containing our new part termed as BBa_K2559003under a strong E.coli endogenous promoter (PrplJ). Therefore, the amended eGFP in BBa_K2559005 was driven by PrplJ promoter, and expressed in DH10B. In addition, we also applied the BBa_K2559005 in the promoter intensity analysis of our other two new parts, the BBa_K2559004 and BBa_K2559011 which are relatively weaker E.coli endogenous promoters (PdapA and PcaiF) (Figure 1).
Figure 1: Fluorescent intensity of amended eGFP driven by by PrplJ, PdapA, PcaiF promoter.
We summarized that our improvedpart, the amended eGFP coding biobrick BBa_K2559005 worked well in DH10B. We also hoped that our improvement on the BBa_I714891 can help their future applications by other groups in the future. However, it is difficult for us to perform additional experiments with BBa_K2559005 and BBa_I714891 due to the unavailable BBa_I714891.
To expand the application of BBa_K2559005, we searched theBBa_J04450stored in registry and do another improvement in the BBa_J04450. The BBa_J04450 is a strong RFP expression vector in E.coli. As the main page of BBa_J04450 mentioned, the E.coli colonies with BBa_J04450 were in red color under normal light after about 18 hour culture on LB plate (Figure 2). We used the BBa_K2559005 to replace the RFP region in BBa_J04450, the modified part is BBa_K2559009. We transferred the BBa_K2559009 to DH5α by heat-shock, and found that the fluorescence signal can be observed under the UV (Figure 2).
Figure 2 Colonies with BBa_J04450 (red colony) and BBa_K2559009 (green colony) visualized under UV lightbox.
So, we confirm that our improved part BBa_K2559005 can work in different E.coli expression system. We are also looking forward to more application of the BBa_K2559009!
School's name:SCAU
Member's name:SCAU
Designed by:SCAU