JimmyZhong (Talk | contribs) |
JimmyZhong (Talk | contribs) |
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{{iTesla-SoundBio}} | {{iTesla-SoundBio}} | ||
<html> | <html> | ||
+ | <center> | ||
+ | <h1>Build the change<br> | ||
+ | <script> | ||
+ | $("#title").text("Project Description") | ||
+ | |||
+ | </script> | ||
+ | <br>You want to see in the world</h1> | ||
+ | </center> | ||
+ | |||
+ | |||
+ | <script src="https://ajax.googleapis.com/ajax/libs/jquery/3.3.1/jquery.min.js"></script> | ||
+ | <script> | ||
+ | $(document).ready(function(){ | ||
+ | // Add smooth scrolling to all links | ||
+ | $("a").on('click', function(event) { | ||
+ | |||
+ | // Make sure this.hash has a value before overriding default behavior | ||
+ | if (this.hash !== "") { | ||
+ | // Prevent default anchor click behavior | ||
+ | event.preventDefault(); | ||
+ | |||
+ | // Store hash | ||
+ | var hash = this.hash; | ||
+ | |||
+ | // Using jQuery's animate() method to add smooth page scroll | ||
+ | // The optional number (800) specifies the number of milliseconds it takes to scroll to the specified area | ||
+ | $('html, body').animate({ | ||
+ | scrollTop: $(hash).offset().top | ||
+ | }, 800, function(){ | ||
+ | |||
+ | // Add hash (#) to URL when done scrolling (default click behavior) | ||
+ | window.location.hash = hash; | ||
+ | }); | ||
+ | } // End if | ||
+ | }); | ||
+ | }); | ||
+ | </script> | ||
+ | |||
<style> | <style> | ||
+ | body { | ||
+ | font-family: "Lato", sans-serif; | ||
+ | } | ||
+ | |||
+ | .sidenav { | ||
+ | height: 80%; | ||
+ | width: 180px; | ||
+ | position: fixed; | ||
+ | z-index: 1; | ||
+ | top: 0; | ||
+ | left: 0; | ||
+ | background-color: #111; | ||
+ | overflow-x: hidden; | ||
+ | padding-top: 200px; | ||
+ | } | ||
+ | |||
+ | .sidenav a { | ||
+ | padding: 5px 5px 5px 5px; | ||
+ | text-decoration: none; | ||
+ | font-size: 20px; | ||
+ | color: #818181; | ||
+ | display: block; | ||
+ | } | ||
+ | |||
+ | .sidenav a:hover { | ||
+ | color: #f1f1f1; | ||
+ | } | ||
+ | |||
+ | .main { | ||
+ | margin-left: 180px; /* Same as the width of the sidenav */ | ||
+ | padding: 0px 10px; | ||
+ | font-family: arial; | ||
+ | font-size: 20px; | ||
+ | padding: 30px | ||
+ | } | ||
+ | @media screen and (max-height: 450px) { | ||
+ | .sidenav {padding-top: 15px;} | ||
+ | .sidenav a {font-size: 18px;} | ||
+ | } | ||
+ | .graphicsum { | ||
+ | width: 100%; | ||
+ | margin: auto; | ||
+ | } | ||
+ | .text { | ||
+ | font-size: 20px; | ||
+ | letter-spacing: .16em; | ||
+ | padding-left: 10px; | ||
+ | padding-right: 100px; | ||
+ | padding-top: 10px; | ||
+ | padding-bottomL 10px; | ||
+ | color: white; | ||
+ | |||
+ | } | ||
+ | |||
h1 { | h1 { | ||
font: 600 1.5em/1 'Raleway', sans-serif; | font: 600 1.5em/1 'Raleway', sans-serif; | ||
Line 39: | Line 131: | ||
-webkit-mask-image: url(https://f.cl.ly/items/1t1C0W3y040g1J172r3h/mask.png); | -webkit-mask-image: url(https://f.cl.ly/items/1t1C0W3y040g1J172r3h/mask.png); | ||
} | } | ||
− | + | .horizontalline { | |
+ | border: 2px solid black; | ||
+ | width: 50%; | ||
+ | height: 1 px; | ||
+ | margin: auto; | ||
+ | margin-top: 50px; | ||
+ | margin-bottom: 10px; | ||
+ | } | ||
</style> | </style> | ||
+ | |||
<body> | <body> | ||
− | + | ||
+ | |||
+ | |||
+ | <div class="sidenav"> | ||
+ | <a href="#graphicsum">Graphic Summary</a> | ||
<br> | <br> | ||
− | + | <a href="#constructdesign">Construct Design</a> | |
+ | <br> | ||
+ | <a href="#Biobrick">Biobrick: <br>Factor C gene in E. coli pSB1C3</a> | ||
+ | <br> | ||
+ | <a href="#Integration">Integration: <br>Factor C in B.Subtilis Chromosomal DNA</a> | ||
+ | <br> | ||
+ | <a href="#Expression">Expression: <br>Factor C protein verification</a> | ||
+ | <br> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <div class="main"> | ||
+ | <div id="graphicsum"> | ||
+ | <center> | ||
+ | <img src="https://static.igem.org/mediawiki/2018/9/9b/T--iTesla-Soundbio--Project_Description1.png" alt="DID YOU KNOW? Every year, hundreds of thousands of horseshoe crabs are caught each year and drained of up to to 30% of their blood. Why? To collect LAL, a chemical that is crucial in the detection of endotoxins in everything from drugs to medical equipment", width=80%> | ||
+ | </center> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | <div class="horizontalline"> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | <div id="constructdesign"> | ||
+ | <p class="text"> | ||
+ | Construct design: <br> | ||
+ | The goal for our project is to insert Factor C gene into the chromosomal DNA of B. Subtilis, and have it express this Factor C protein. At the same time, to fulfill iGEM competition requirements, we also need to insert Factor C gene into E.coli plasmid pSB1C3 and submit it as a biobrick for iGEM competition. | ||
+ | <br> | ||
+ | We decided to insert Factor C gene into E.coli plasmid pSB1C3 first; then, integrate Factor C gene into the chromosomal DNA of B. Subtilis using pAX01 integration factor (a B. Subtilis specific plasmid). | ||
+ | </P> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | <div class="horizontalline"> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <div id="Biobrick"> | ||
+ | <center><img src="https://static.igem.org/mediawiki/2018/c/c6/T--itesla-soundbio--twofragments.png", width=80%></center> | ||
+ | <p class="text"> | ||
+ | Biobrick: <br> | ||
+ | Factor C in E.coli pSB1C3 | ||
+ | We couldn't order Factor C from factory because it is too large (about 3000 base pairs https://benchling.com/itesla-soundbio/f/gPUeHZXf-drafts/seq-Jq87odxl-psb1c3factor-c/edit), so we order it as two fragments, FC1 and FC2. | ||
+ | First, insert FC1 into pSB1C3. Then, insert FC2 into the plasmid, too. This pSB1C3 with full Factor C will be submitted for biobrick. | ||
+ | </p> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | <div class="horizontalline"> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | <div id="Integration"> | ||
+ | <p class="text"> | ||
+ | Integration: <br> | ||
+ | Factor C in B. Subtilis Chromosomal DNA. | ||
+ | We then digest full Factor C from Biobrick, and insert it into pAX01, a B. Subtilis integration factor. Then, we transform B. Subtilis with pAX01. After plasmid pAX01 enters B. Subtilis, there is a chance that it will integrate into chromosomal DNA. | ||
+ | </p> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | <div class="horizontalline"> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <div id="expression"> | ||
+ | <p class="text"> | ||
+ | Expression | ||
+ | Factor C protein extract: <br> | ||
+ | B. Subtilis are used to express Factor C because they are gram positive and will not damage the Factor C protein. In contrast, E. coli is gram negative and will cleave FC protein. After B. Subtilis express and produce Factor C, we can then extract Factor C protein for further experiment. | ||
+ | </p> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | </div> | ||
+ | </div> | ||
</body> | </body> | ||
+ | |||
+ | |||
+ | |||
</html> | </html> | ||
+ | |||
+ | <!-- | ||
+ | #boxshadow { | ||
+ | position: relative; | ||
+ | box-shadow: 1px 2px 4px rgba(0, 0, 0, .5); | ||
+ | padding: 10px; | ||
+ | background: white; | ||
+ | } | ||
+ | |||
+ | #boxshadow img { | ||
+ | width: 100%; | ||
+ | border: 1px solid #8a4419; | ||
+ | border-style: inset; | ||
+ | } | ||
+ | |||
+ | #boxshadow::after { | ||
+ | content: ''; | ||
+ | position: absolute; | ||
+ | z-index: -1; /* hide shadow behind image */ | ||
+ | box-shadow: 0 15px 20px rgba(0, 0, 0, 0.3); | ||
+ | width: 70%; | ||
+ | left: 15%; /* one half of the remaining 30% */ | ||
+ | height: 100px; | ||
+ | bottom: 0; | ||
+ | } | ||
+ | background-color: #008CBA; | ||
+ | border: none; | ||
+ | color: white; | ||
+ | padding: 15px 32px; | ||
+ | text-align: center; | ||
+ | text-decoration: none; | ||
+ | display: inline-block; | ||
+ | font-size: 16px; | ||
+ | margin: 4px 2px; | ||
+ | cursor: pointer; | ||
+ | display: flex; justify-content: center; | ||
+ | } | ||
+ | --> |
Revision as of 03:03, 30 September 2018
Build the change
You want to see in the world
Construct Design
Biobrick:
Factor C gene in E. coli pSB1C3
Integration:
Factor C in B.Subtilis Chromosomal DNA
Expression:
Factor C protein verification
Construct design:
The goal for our project is to insert Factor C gene into the chromosomal DNA of B. Subtilis, and have it express this Factor C protein. At the same time, to fulfill iGEM competition requirements, we also need to insert Factor C gene into E.coli plasmid pSB1C3 and submit it as a biobrick for iGEM competition.
We decided to insert Factor C gene into E.coli plasmid pSB1C3 first; then, integrate Factor C gene into the chromosomal DNA of B. Subtilis using pAX01 integration factor (a B. Subtilis specific plasmid).
Biobrick:
Factor C in E.coli pSB1C3
We couldn't order Factor C from factory because it is too large (about 3000 base pairs https://benchling.com/itesla-soundbio/f/gPUeHZXf-drafts/seq-Jq87odxl-psb1c3factor-c/edit), so we order it as two fragments, FC1 and FC2.
First, insert FC1 into pSB1C3. Then, insert FC2 into the plasmid, too. This pSB1C3 with full Factor C will be submitted for biobrick.
Integration:
Factor C in B. Subtilis Chromosomal DNA.
We then digest full Factor C from Biobrick, and insert it into pAX01, a B. Subtilis integration factor. Then, we transform B. Subtilis with pAX01. After plasmid pAX01 enters B. Subtilis, there is a chance that it will integrate into chromosomal DNA.
Expression
Factor C protein extract:
B. Subtilis are used to express Factor C because they are gram positive and will not damage the Factor C protein. In contrast, E. coli is gram negative and will cleave FC protein. After B. Subtilis express and produce Factor C, we can then extract Factor C protein for further experiment.