Difference between revisions of "Part:BBa K1228008"

(→‎Experiment)
(→‎Experiment)
 
(73 intermediate revisions by 3 users not shown)
Line 2: Line 2:
 
<partinfo>BBa_K1228008 short</partinfo>
 
<partinfo>BBa_K1228008 short</partinfo>
  
−
Human lysozyme (hLZ), the actual protein in breast milk,exhibits microbicidal activity to various degrees against several bacterial strains.The HLH peptide and its N-terminal helix (H1) were significantly the most potent bactericidal to Gram-positive and Gram-negative bacteria.Evidence shows that HLH peptide and its N-terminal helix (H1) kill bacteria by crossing the outer membrane of Gram-negative bacteria via self-promoted uptake and are able to dissipate the membrane potential-dependent respiration of Gram-positive bacteria.
+
Human Llysozyme (hLZ), the major protein in breast milk, exhibits microbicidal activity to various degrees against several bacterial strains. This Generator can secrete the HLH peptide(38 aminoacid peptides) that is a part of Human Lysozyme in ''B.subtilis''. The HLH peptide and its N-terminal helix (H1) were the most potent bactericidal to bacteria. Evidence shows that HLH peptide and its N-terminal helix (H1) kill bacteria by crossing the outer membrane of Gram-negative bacteria via self-promoted uptake and is able to dissipate the membrane-potential-dependent respiration of Gram-positive bacteria.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 19: Line 19:
  
 
== Experiment ==
 
== Experiment ==
−
In order to distinguish whether the ''B. Subtilis''express our objective protein or not,we used the method of in-vitro bacterial inhibition zone to observe the anti-bacterial effect.We added 100 ul ''B. Subtilis'' liquid medium into the hole. The inhibition zone results showed that gram-positive bacterium was sensitive to 38aa.
+
In order to distinguish whether the ''B. Subtilis'' express our objective protein or not,we used the method of in-vitro bacterial inhibition zone to observe the anti-bacterial effect.We added 100 ul ''B. Subtilis'' liquid medium into the hole. The inhibition zone results showed that gram-positive bacterium was sensitive to 38aa.
  
−
https://static.igem.org/mediawiki/2013/0/0a/L2_%E6%8A%91%E8%8F%8C%E5%9C%88_Mly.jpg
 
−
 
−
{{:Team：HZAU-China/background-head}}
 
 
<html>
 
<html>
−
<script type="text/javascript" src="/ajaxjs/jquery1.3.2.js"></script>
+
<head>
 +
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
 
<style type="text/css">
 
<style type="text/css">
−
#tip  {position:absolute;color:#333;display:none;z-index:1000;}
+
.zoomPan{width:800px;position:relative; }
−
#tip s  {position:absolute;top:40px;left:-20px;display:block;width:0px;height:0px;font-size:0px;line-height:0px;border-color:transparent #BBA transparent transparent;border-style:dashed solid dashed dashed;border-width:10px;z-index:1000;}
+
.sh{zoom:1;background:url(https://static.igem.org/mediawiki/2013/8/80/Old_wall.jpg);filter:progid:DXImageTransform.Microsoft.dropShadow(color='#54000000', OffX=2,OffY=2);-webkit-box-shadow:4px 4px 4px #666;-moz-box-shadow:4px 4px 4px #666;}
−
#tip s i  {position:absolute;top:-10px;left:-8px;display:block;width:0px;height:0px;font-size:0px;line-height:0px;border-color:transparent #fff transparent transparent;border-style:dashed solid dashed dashed;border-width:10px;z-index:1000;}
+
#zoom{position:absolute;width:254px;height:254px;border:3px solid #fff;left:-9999px;top:0;overflow:hidden;background:#fff;}
−
#tip .t_box  {position:relative;background-color:#CCC;filter:alpha(opacity=50);-moz-opacity:0.5;bottom:-3px;right:-3px;z-index:1000;}
+
#zoom img{position:relative;}
−
#tip .t_box div  {position:relative;background-color:#FFF;border:1px solid #ACA899;background:#FFF;padding:1px;top:-3px;left:-3px;z-index:1000;}
+
−
+
−
.tip  {width:82px;height:82px;}
+
 
</style>
 
</style>
 +
</head>
 +
 
<script type="text/javascript">
 
<script type="text/javascript">
−
$(function(){
+
//<![CDATA[
−
  $('.tip').mouseover(function(){
+
function zoomBox() {this.index.apply(this, arguments)}
−
      var $tip=$('<div id="tip"><div class="t_box"><div><s><i></i></s><img src="'+this.src+'" /></div></div></div>');
+
zoomBox.prototype = {
−
      $('body').append($tip);
+
    index: function(win,zoom) {
−
      $('#tip').show('fast');
+
        var win=document.getElementById(win);
−
  }).mouseout(function(){
+
        var box=document.getElementById(zoom);
−
      $('#tip').remove();
+
        var img=box.getElementsByTagName('IMG')[0];
−
  }).mousemove(function(e){
+
        var zoom=img.width/win.getElementsByTagName('IMG')[0].width;
−
      $('#tip').css({"top":(e.pageY-60)+"px","left":(e.pageX+30)+"px"})
+
        var z=Math.round(box.offsetWidth/2);
−
  })
+
        win.onmousemove=function (e){
−
})
+
            e = e || window.event;
 +
            var x=e.clientX,y=e.clientY, ori=win.getBoundingClientRect();
 +
            if (x>ori.right+20||y>ori.bottom+20||x<ori.left-20||y<ori.top-20)box.style.display='none';
 +
            x-=ori.left;
 +
            y-=ori.top;
 +
            box.style.left=x-z+'px';
 +
            box.style.top=y-z+'px';
 +
            img.style.left=-x*zoom+z+'px';
 +
            img.style.top=-y*zoom+z+'px';
 +
        }
 +
        win.onmouseover=function (){box.style.display=''}
 +
    }
 +
};
 +
window.onload=function (){
 +
    x=new zoomBox('zoomPan','zoom')
 +
}
 +
//]]>
 
</script>
 
</script>
  
−
<style>
+
   
−
*{margin:0px;padding:0px;}
+
−
ul.menu_scroll{z-index:100;padding:10px;margin:0 auto;padding-bottom: 5px;top:20px;position:fixed;width:100%;}
+
−
ul.menu:after{content:"";display:block;height:0;clear:both;visibility:hidden;}
+
−
ul.menu,
+
−
ul.menu ul {padding:0;margin:0;list-style:none;position:;width:220px;background:#ddd;font-family:arial, sans-serif;}
+
−
ul.menu {z-index:100;padding:10px;margin:0 auto;}
+
−
ul.menu ul {z-index:50;
+
−
-webkit-transition: 0.5s;
+
−
-moz-transition: 0.5s;
+
−
-ms-transition: 0.5s;
+
−
-o-transition: 0.5s;
+
−
transition: 0.5s;
+
−
}
+
−
ul.menu li {line-height:30px;position:relative;}
+
−
ul.menu > li {margin-top:2px;font-size:12px;}
+
−
ul.menu > li a {font:normal 16px/29px arial, sans-serif;color:#fff;text-decoration:none;}
+
−
ul.menu label.open {display:block;background:#c00 no-repeat 170px 12px;line-height:30px;position:relative;z-
+
  
−
index:100;font:normal 12px/30px arial, sans-serif;color:#fff;border-radius:10px 10px 0 0;}
+
<div id="paragraphs">
−
ul.menu label.open b {color:#ff0;}
+
        <div class="zoomPan" id="zoomPan">
−
ul.menu span {display:block;background:#c00;line-height:30px;position:relative;z-index:100;font-size: 16px;padding-left:10px;}
+
         <img src="https://static.igem.org/mediawiki/2013/0/0a/L2_%E6%8A%91%E8%8F%8C%E5%9C%88_Mly.jpg" width="800px;">
−
ul.menu label img {position:absolute;left:0;top:0;width:100%;height:30px;}
+
         <div id="zoom" class="sh"><img src="https://static.igem.org/mediawiki/igem.org/a/a8/Yijunquanend.jpg" width="2000px;"></div>
−
ul.menu ul li {margin-top:-30px;
+
        </div>
−
-webkit-transition: 0.5s;
+
−
-moz-transition: 0.5s;
+
−
-ms-transition: 0.5s;
+
−
-o-transition: 0.5s;
+
−
transition: 0.5s;
+
−
}
+
−
ul.menu ul li a {display:block;font:normal 16px/29px arial, sans-serif;color:#000;background:#ccc;}
+
−
ul.menu ul li a:hover {background:#ddd;}
+
−
ul.menu input {position:absolute;left:-9999px;}
+
−
ul.menu li input:checked + label {background:#069;}
+
−
ul.menu li input:checked ~ ul {background:#ccc;padding-bottom:10px;}
+
−
ul.menu li input:checked ~ ul li {margin-top:0;}
+
−
ul.menu label.close {display:block;width:200px;height:30px;background:transparent url("/jscss/demoimg/201208/u-arrow.gif")
+
−
 
+
−
no-repeat 170px 12px;line-height:30px;position:relative;left:0;z-index:90;margin-top:-30px}
+
−
ul.menu input.tabs:checked ~ label.close {display:block;z-index:200;}
+
−
ul.menu input.close:checked + label.close {display:none;}
+
−
ul.menu input.close:checked ~ ul {background:#ddd;padding-bottom:0;}
+
−
ul.menu input.close:checked ~ ul li {margin-top:-30px;}
+
−
</style>
+
−
 
+
−
<script>
+
−
$(document).ready(function(){
+
−
var topMain=$("#sidebar").height()+100//是头部的高度加头部与nav导航之间的距离
+
−
var menu=$(".menu");
+
−
$(window).scroll(function(){
+
−
if ($(window).scrollTop()>topMain){//如果滚动条顶部的距离大于topMain则就nav导航就添加类.nav_scroll，否则就移除
+
−
menu.addClass("menu_scroll");
+
−
}else{
+
−
menu.removeClass("menu_scroll");
+
−
}
+
−
});
+
−
})
+
−
</script>
+
−
 
+
−
<body>
+
−
<div id="background">
+
−
    <div id="sidebar">
+
−
        <body>
+
−
        <ul class="menu">
+
−
         <li><a href="http://2013.igem.org/Team:HZAU-China/Meeting Record"><span>Meeting Record</span></a></li>
+
−
         <li><a href="http://2013.igem.org/Team:HZAU-China/Gallery"><span style="font-size:19px;color=#fff;">Gallery</span></a></li>
+
−
        <li><a href="http://2013.igem.org/Team:HZAU-China/Protocol"><span>Protocol</span></a></li>
+
−
      </body>
+
 
     </div>
 
     </div>
−
    <div id="paragraphs">
 
−
    <p><br></p>
 
−
    <center><span style="font-size:46px;font-family:Cambria;margin-top:10px;line-height:80%">Gallery</span></center>
 
−
    <p><br></p>
 
−
   
 
−
<p> </p>
 
−
<a><img class="tip" src="https://static.igem.org/mediawiki/2013/0/0a/L2_%E6%8A%91%E8%8F%8C%E5%9C%88_Mly.jpg" /></a>
 
−
 
−
    </div>
 
−
</div>
 
−
</body>
 
 
</html>
 
</html>
  
−
{{:Team：HZAU-China/background-footer}}
+
https://static.igem.org/mediawiki/2013/9/97/ZswyjqMly02s.jpg
−
<br/>
+
  
 
== Reference ==
 
== Reference ==
 +
1.Hisham R. Ibrahim,* Kenta Imazato, and Hajime Ono. Human Lysozyme Possesses Novel Antimicrobial Peptides within Its
 +
N-terminal Domain that Target Bacterial Respiration. J Agric Food Chem. 2011 Sep 28;59(18):10336-45. doi: 10.1021/jf2020396. Epub 2011 Sep 2.

Latest revision as of 12:03, 27 September 2013

38aa with T1 terminator and Constitutive promoter veg

Human Llysozyme (hLZ), the major protein in breast milk, exhibits microbicidal activity to various degrees against several bacterial strains. This Generator can secrete the HLH peptide(38 aminoacid peptides) that is a part of Human Lysozyme in B.subtilis. The HLH peptide and its N-terminal helix (H1) were the most potent bactericidal to bacteria. Evidence shows that HLH peptide and its N-terminal helix (H1) kill bacteria by crossing the outer membrane of Gram-negative bacteria via self-promoted uptake and is able to dissipate the membrane-potential-dependent respiration of Gram-positive bacteria.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



Experiment

In order to distinguish whether the B. Subtilis express our objective protein or not,we used the method of in-vitro bacterial inhibition zone to observe the anti-bacterial effect.We added 100 ul B. Subtilis liquid medium into the hole. The inhibition zone results showed that gram-positive bacterium was sensitive to 38aa.

ZswyjqMly02s.jpg

Reference

1.Hisham R. Ibrahim,* Kenta Imazato, and Hajime Ono. Human Lysozyme Possesses Novel Antimicrobial Peptides within Its N-terminal Domain that Target Bacterial Respiration. J Agric Food Chem. 2011 Sep 28;59(18):10336-45. doi: 10.1021/jf2020396. Epub 2011 Sep 2.