Line 603: | Line 603: | ||
<!------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- Content begins ---------------------------------------------------------------------------------------------------------------------------------> | <!------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- Content begins ---------------------------------------------------------------------------------------------------------------------------------> | ||
<!----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------> | <!----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------> | ||
− | + | ||
Line 789: | Line 789: | ||
color:white; | color:white; | ||
display:block; | display:block; | ||
+ | margin-top:10%; | ||
+ | margin-bottom:7%; | ||
} | } | ||
.title2{ | .title2{ | ||
Line 809: | Line 811: | ||
.para1{ | .para1{ | ||
color:white; | color:white; | ||
− | text-align:justify; | + | text-align:justify !important; |
align-items:flex-start; | align-items:flex-start; | ||
line-height:140%; | line-height:140%; | ||
Line 849: | Line 851: | ||
display:block; | display:block; | ||
margin-left:25%; | margin-left:25%; | ||
+ | } | ||
+ | .legend{ | ||
+ | color:white; | ||
+ | text-align:center; | ||
+ | } | ||
+ | .legend>div{ | ||
+ | width:100%; | ||
+ | text-align:center!important; | ||
+ | } | ||
+ | .legends{ | ||
+ | margin:auto 15%; | ||
} | } | ||
Line 1,323: | Line 1,336: | ||
</li> | </li> | ||
− | <li class="navLi"><a class="navA noSubmenu" href="https:// | + | <li class="navLi"><a class="navA noSubmenu" href="https://igem.org/2019_Judging_Form?id=3257">Judging</a></li> |
</ul> | </ul> | ||
Line 1,376: | Line 1,389: | ||
#demoCover{ | #demoCover{ | ||
width:100vw; | width:100vw; | ||
− | height: | + | height:80vh; |
position:absolute; | position:absolute; | ||
background-color:rgba(8,39,58,0.5); | background-color:rgba(8,39,58,0.5); | ||
− | top: | + | top:70px; |
left:0; | left:0; | ||
text-align:center; | text-align:center; | ||
} | } | ||
#coverPic{ | #coverPic{ | ||
− | width: | + | width:550px; |
− | margin: | + | margin:20vh auto; |
} | } | ||
#demo{ | #demo{ | ||
width:100vw; | width:100vw; | ||
− | height: | + | height:70vh; |
position:relative; | position:relative; | ||
} | } | ||
Line 1,411: | Line 1,424: | ||
#demoCover{ | #demoCover{ | ||
top:55px; | top:55px; | ||
+ | height:30vh; | ||
} | } | ||
#demo{ | #demo{ | ||
Line 1,416: | Line 1,430: | ||
} | } | ||
#coverPic{ | #coverPic{ | ||
− | width: | + | width:500px; |
margin:7vh auto; | margin:7vh auto; | ||
} | } | ||
Line 1,428: | Line 1,442: | ||
} | } | ||
#coverPic{ | #coverPic{ | ||
− | width: | + | width:500px; |
margin:6vh auto; | margin:6vh auto; | ||
} | } | ||
Line 1,440: | Line 1,454: | ||
} | } | ||
#coverPic{ | #coverPic{ | ||
− | width: | + | width:400px; |
+ | margin:8vh auto; | ||
+ | } | ||
+ | } | ||
+ | @media only screen and (max-width:500px){ | ||
+ | #coverPic{ | ||
+ | width:200px; | ||
margin:8vh auto; | margin:8vh auto; | ||
} | } | ||
Line 1,782: | Line 1,802: | ||
<ul class="leftNav" style="margin:0;padding:0;"> | <ul class="leftNav" style="margin:0;padding:0;"> | ||
− | <li class="leftNavLi"><a class="leftNavA" href="#mainTitle1"> | + | <li class="leftNavLi"><a class="leftNavA" href="#mainTitle1">Reverse transcription</a> |
<ul class="leftNavUl2"> | <ul class="leftNavUl2"> | ||
− | <li class="leftNavLi2"><a class="leftNavA2" href="#mainTitle1_1"> | + | <li class="leftNavLi2"><a class="leftNavA2" href="#mainTitle1_1">MMLV RT</a></li> |
<li class="leftNavLi2"><a class="leftNavA2" href="#mainTitle1_2">Reverse transcriptase</a></li> | <li class="leftNavLi2"><a class="leftNavA2" href="#mainTitle1_2">Reverse transcriptase</a></li> | ||
<li class="leftNavLi2"><a class="leftNavA2" href="#mainTitle1_3">Anti-microbial resistance (AMR)</a></li> | <li class="leftNavLi2"><a class="leftNavA2" href="#mainTitle1_3">Anti-microbial resistance (AMR)</a></li> | ||
Line 1,896: | Line 1,916: | ||
+ | <div class="row title1" id="mainTitle1"> | ||
+ | <div class="col">Reverse transcription</div> | ||
+ | </div> | ||
+ | |||
+ | <div class="row title2" id="mainTitle1_1"> | ||
+ | <div class="col">Moloney murine leukemia virus reverse transcriptase</div> | ||
+ | </div> | ||
+ | |||
+ | <div class="row title3"> | ||
+ | <div class="col"><i>Why MMLV?</i></div> | ||
+ | </div> | ||
+ | |||
<div class="row para1"> | <div class="row para1"> | ||
<div class="row"> | <div class="row"> | ||
− | <div class="col"> | + | <div class="col col-md-12"> |
− | + | Reverse transcriptase (RT) is one of the most crucial part in our system, we chose it from Moloney murine leukemia virus (MMLV) for five reasons. | |
</div> | </div> | ||
− | </div> | + | </div> |
+ | <div class="row"> | ||
+ | <div class="col"> | ||
+ | <ul class="paraUl" style="list-style:none;"> | ||
+ | <li>MMLV-RT’s enhanced version is commonly used in in vitro reverse transcription, which guarantees the safety and well characterization of this part.</li> | ||
+ | <li>Unlike the reverse transcriptase of HIV, MMLV-RT acts as a monomer, this brings less trouble to its production in an heterogenous host.</li> | ||
+ | <li>MMLV is a eukaryotic virus, which means it’s orthogonal to prokaryotic species. This orthogonality makes up of the host adaptability of our system.</li> | ||
+ | <li>MMLV-RT has a higher processivity in relation to other viruses’ RT such as HIV and AMV. This feature gives our system a greater range of mutagenesis length.</li> | ||
+ | <li>MMLV-RT has low primer specificity, which means that if we change the sequence of its primer and its corresponding primer binding site (PBS), unlike HIV, the change will not be reverted in the following reverse transcription process. This makes it possible for us to customize the sequence of the tRNA primer used in accord with the target sequence in our software.</li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | </div> | ||
</div> | </div> | ||
− | <div class="row | + | <div class="row title3"> |
− | <div class="col">Design</div> | + | <div class="col"><i>Design</i></div> |
</div> | </div> | ||
− | |||
− | |||
− | |||
− | |||
<div class="row para1"> | <div class="row para1"> | ||
<div class="row"> | <div class="row"> | ||
<div class="col"> | <div class="col"> | ||
− | + | Even though RT does the function of reverse transcription, what is expressed in the cell is its polyprotein version. The gag-pol polyprotein has four parts—capsid protein, protease (stop codon separated), reverse transcriptase, and integrase. The integrase is deleted from the polyprotein to eliminate the possibility of genome interference. The protease contains a UAG stop codon at its 5th amino acid site1, which is readthrough as glutamine at a 5% efficiency in its native host cells, to enable the 20:1 ratio between capsid and reverse transcriptase protein. As its readthrough efficiency is much lower in E. coli cells, and studies have shown that lower efficiency greatly damages the activity of reverse transcriptase2, we mutated the UAG codon into CAG, making a complete readthrough of the polyprotein. This will slightly decrease the activity of RT but within an acceptable range. The capsid protein is necessary as it has been found to promote the annealing of tRNA primer to the primer binding site (PBS) in MMLV, and plays an important role in the following two strand transfer steps3,4. To be certain of this design, we have consulted Prof. Alper through mail and received his confirmation on the necessity of the capsid protein.<br /><br /> | |
+ | To increase the mutation of our RT, we built a mutated version (Y1245F). This mutation has been shown to increase the mutation level by 5 times5.<br /><br /> | ||
+ | The gag-pol polyprotein is placed under Lac operon, whose expression controlled by IPTG.<br /><br /> | ||
+ | |||
</div> | </div> | ||
</div> | </div> | ||
Line 2,046: | Line 2,088: | ||
<div class="col-sm-4"> | <div class="col-sm-4"> | ||
− | <div class="footerUl align-items-center | + | <div class="footerUl align-items-center"> |
<span style="text-decoration:none;">Model</span> | <span style="text-decoration:none;">Model</span> | ||
<ul> | <ul> | ||
Line 2,074: | Line 2,116: | ||
<div class="col-sm-4"> | <div class="col-sm-4"> | ||
− | <div class="footerUl align-items-center"> | + | <div class="footerUl align-items-center active"> |
<span><a href="/Team:Fudan-TSI/Human_Practices" style="text-decoration:none;">Outreach</a></span> | <span><a href="/Team:Fudan-TSI/Human_Practices" style="text-decoration:none;">Outreach</a></span> | ||
<ul> | <ul> | ||
Line 2,132: | Line 2,174: | ||
var winHeight=$(window).height(); | var winHeight=$(window).height(); | ||
var winWidth=$(window).width(); | var winWidth=$(window).width(); | ||
− | var pagePosition=$("#pageContent").offset().top; | + | var pagePosition=$("#pageContent").offset().top+90; |
var navHeight=$(".leftNav").height(); | var navHeight=$(".leftNav").height(); | ||
var footerHeight=$("#FudanFooter").height(); | var footerHeight=$("#FudanFooter").height(); | ||
− | var maxTopDistance=$("# | + | var maxTopDistance=$("#FudanFooter").offset().top-$(".leftNav").height()-$("#top_menu_14").height()-$("#navUl").height(); |
$(".leftNav").css({"position":"absolute","top":pagePosition}); | $(".leftNav").css({"position":"absolute","top":pagePosition}); | ||
$(window).resize(function(){ | $(window).resize(function(){ | ||
var winHeight=$(window).height(); | var winHeight=$(window).height(); | ||
var winWidth=$(window).width(); | var winWidth=$(window).width(); | ||
− | var topDistance=$(window).scrollTop(); | + | var topDistance=$(window).scrollTop()+0.25*winHeight; |
var navHeight=$(".leftNav").height(); | var navHeight=$(".leftNav").height(); | ||
var footerHeight=$("#FudanFooter").height(); | var footerHeight=$("#FudanFooter").height(); | ||
− | var maxTopDistance=$("# | + | var maxTopDistance=$("#FudanFooter").offset().top-$(".leftNav").height()-$("#navUl").height(); |
var leftNavFromTop=pagePosition-$(window).scrollTop(); | var leftNavFromTop=pagePosition-$(window).scrollTop(); | ||
− | if (leftNavFromTop<0. | + | if (leftNavFromTop<0.25*winHeight){ |
if(topDistance<maxTopDistance){ | if(topDistance<maxTopDistance){ | ||
− | $(".leftNav").css({"position":"fixed","top":0. | + | $(".leftNav").css({"position":"fixed","top":0.25*winHeight+$("#top_menu_14").height()}); |
} | } | ||
else{ | else{ | ||
Line 2,162: | Line 2,204: | ||
var winWidth=$(window).width(); | var winWidth=$(window).width(); | ||
var leftNavFromTop=pagePosition-$(window).scrollTop(); | var leftNavFromTop=pagePosition-$(window).scrollTop(); | ||
− | var topDistance=$(window).scrollTop(); | + | var topDistance=$(window).scrollTop()+0.25*winHeight; |
− | if (leftNavFromTop<0. | + | if (leftNavFromTop<0.25*winHeight){ |
if(topDistance<maxTopDistance){ | if(topDistance<maxTopDistance){ | ||
− | $(".leftNav").css({"position":"fixed","top":0. | + | $(".leftNav").css({"position":"fixed","top":0.25*winHeight+$("#top_menu_14").height()}); |
} | } | ||
else{ | else{ | ||
Line 2,218: | Line 2,260: | ||
}); | }); | ||
− | </script> | + | </script> |
Revision as of 19:21, 14 October 2019