Difference between revisions of "Part:BBa K1228008"

(→‎Experiment)
(→‎Experiment)
 
(56 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.
 +
 
 +
<html>
 +
<head>
 +
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
 +
<style type="text/css">
 +
.zoomPan{width:800px;position:relative; }
 +
.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;}
 +
#zoom{position:absolute;width:254px;height:254px;border:3px solid #fff;left:-9999px;top:0;overflow:hidden;background:#fff;}
 +
#zoom img{position:relative;}
 +
</style>
 +
</head>
 +
 
 +
<script type="text/javascript">
 +
//<![CDATA[
 +
function zoomBox() {this.index.apply(this, arguments)}
 +
zoomBox.prototype = {
 +
    index: function(win,zoom) {
 +
        var win=document.getElementById(win);
 +
        var box=document.getElementById(zoom);
 +
        var img=box.getElementsByTagName('IMG')[0];
 +
        var zoom=img.width/win.getElementsByTagName('IMG')[0].width;
 +
        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>
 +
 
 +
 +
 
 +
<div id="paragraphs">
 +
        <div class="zoomPan" id="zoomPan">
 +
        <img src="https://static.igem.org/mediawiki/2013/0/0a/L2_%E6%8A%91%E8%8F%8C%E5%9C%88_Mly.jpg" width="800px;">
 +
        <div id="zoom" class="sh"><img src="https://static.igem.org/mediawiki/igem.org/a/a8/Yijunquanend.jpg" width="2000px;"></div>
 +
        </div>
 +
    </div>
 +
</html>
 +
 
 +
https://static.igem.org/mediawiki/2013/9/97/ZswyjqMly02s.jpg
  
 
== Reference ==
 
== Reference ==
−
Hisham R. Ibrahim,* Kenta Imazato, and Hajime Ono. Human Lysozyme Possesses Novel Antimicrobial Peptides within Its
+
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.
 
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.