Difference between revisions of "Team:SCAU-China/Improve"

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coming soon
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</style>
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<div class="Tan MingYang"></div>
 +
<div class="Li JiaDong"></div>
 +
<div class="Huang XinLing"></div>
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<div class="Fan ZhongZhao"></div>
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<div class="navbar-default">
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<div class="container">
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<div class="navbar-header">
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<a href="https://2018.igem.org/Team:SCAU-China" class="navbar-brand">SCAU-2018</a>
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</div>
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<div class="navbar">
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<ul style="float: left;" class="nav">
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<li class="dropdown">
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<a href="javascript:void(0)">TEAM</a>
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<span class="caret"></span>
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<ul class='dropdown-menu'>
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<li>
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<a href="https://2018.igem.org/Team:SCAU-China/Members">Members</a>
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</li>
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<li>
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<a href="https://2018.igem.org/Team:SCAU-China/Attributions">Attributions</a>
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</ul>
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<li class="dropdown">
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<a href="javascript:void(0)">PROJECT</a>
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<span class="caret"></span>
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<ul class='dropdown-menu'>
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<li><a href="https://2018.igem.org/Team:SCAU-China/ProjectOverview">Overview</a></li>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Background">Background</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Design">Design</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/SRK">Synergistic Recombination Kit</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/MM">Mathematical Model of Biological Intrinsic Regulation System</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Type">Type II CRISPR/Cas 9 Kit</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/MOM">Method for Optimizing Microbial Cell Culture</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Outlook">Outlook</a></li>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Demonstrate">Demonstrate</a></li>
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</ul>
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<li class="dropdown">
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<a href="javascript:void(0)">LAB WORK</a>
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<span class="caret"></span>
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<ul class='dropdown-menu'>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Experiments">Experiments</a></li>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Parts">Parts</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Improve">Improve</a></li>
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<li><a href="https://2018.igem.org/Team:SCAU-China/InterLab">Interlab</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Measurement">Measurement</a></li>
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</ul>
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<li class="dropdown">
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<a href="javascript:void(0)">MODEL</a>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Model">Overview</a></li>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Model/HAWNA">HAWNA</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Model/PPIBoost">PPIBoost</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Model/CultrueCondition">Cultrue Condition</a></li>
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</ul>
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</li>
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</ul>
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<ul style="float: right;" class="nav">
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<li class="dropdown">
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<a href="javascript:void(0)">SAFETY</a>
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<span class="caret"></span>
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<ul class='dropdown-menu'>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Safety">Safety</a></li>
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<li class="dropdown">
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<a href="javascript:void(0)">HUMAN PRACTICES</a>
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<span class="caret"></span>
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<ul class='dropdown-menu'>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Human_Practices">Overview</a></li>
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<li><a href="https://2018.igem.org/Team:SCAU-China/silver">Silver</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Integrated">Integrated</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Public_Engagement">Public Engagement & Education</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Collaborations">Collaborations</a></li>
 +
 
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</ul>
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</li>
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</ul>
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</div>
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</div>
 +
</div>
 +
 
 +
 
 +
<div id="mainDiv">
 +
<div class="heart"><!-- 版心 -->
 +
<div id="mask"><!-- 半透明底板 -->
 +
 
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<div class="DBoard" id="title">
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Improve
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</div>
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<div class="DBoard" id="title2">
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BBa_K2559005 /BBa_K2559009
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</div>
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<div class="DBoard" id="art3">
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<p>
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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>
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<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>
 +
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 +
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();
 +
</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>

Latest revision as of 11:16, 17 October 2018

SCAU-2018
Improve
BBa_K2559005 /BBa_K2559009

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