Difference between revisions of "Team:SCUT/Description"

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{{SCUT}}
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<h2> Project Description </h2>
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<p>Tell us about your project, describe what moves you and why this is something important for your team.</p>
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<h5>What should this page contain?</h5>
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<ul>
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<li> A clear and concise description of your project.</li>
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<li>A detailed explanation of why your team chose to work on this particular project.</li>
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<li>References and sources to document your research.</li>
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<li>Use illustrations and other visual resources to explain your project.</li>
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</ul>
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<h4>Advice on writing your Project Description</h4>
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We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be consist, accurate and unambiguous in your achievements.  
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Judges like to read your wiki and know exactly what you have achieved. This is how you should think about these sections; from the point of view of the judge evaluating you at the end of the year.
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<h4>References</h4>
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<p>iGEM teams are encouraged to record references you use during the course of your research. They should be posted somewhere on your wiki so that judges and other visitors can see how you though about your project and what works inspired you.</p>
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<h4>Inspiration</h4>
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<p>See how other teams have described and presented their projects: </p>
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<ul>
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<meta http-equiv="content-type" content="text/html; charset=utf-8" />  
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<li><a href="https://2014.igem.org/Team:Imperial/Project"> Imperial</a></li>
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<li><a href="https://2014.igem.org/Team:UC_Davis/Project_Overview"> UC Davis</a></li>
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<li><a href="https://2014.igem.org/Team:SYSU-Software/Overview">SYSU Software</a></li>
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<meta name="viewport" content="width=device-width, initial-scale=1, maximum-scale=1">
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</ul>
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</div>
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<title>Team:SCUT/Discription</title>
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<div class="logo"><a href="https://2015.igem.org/Team:SCUT"></a></div>
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<div class="two-third column-last">
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<nav id="navigation">
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<ul id="mainnav">
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<li><a href="https://2015.igem.org/Team:SCUT"><span>HOME</span></a>
 +
 +
</li>
 +
                        <li><a href="#"><span>TEAM</span></a>
 +
                        <ul>
 +
<li><a href="https://2015.igem.org/Team:SCUT/Attributions">Attributions</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/Gallery">Gallery</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/Members">Members</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/OurUniversity">Our University</a></li>
 +
</ul>
 +
 +
</li>
 +
                        <li><a href="#"><span>PROJECT</span></a>
 +
<ul>
 +
 +
<li><a href="https://2015.igem.org/Team:SCUT/BackgroundOverview">Background & Overview</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/CadmiumAbsorption">Cadmium Absorption</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/SensorSystem">Sensor System</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/Protocol">Protocol</a></li>
 +
                                <li><a href="https://2015.igem.org/Team:SCUT/Notebook">NOTEBOOK</a></li>
 +
                                <li><a href="https://2015.igem.org/Team:SCUT/Achievement">ACHIEVEMENT</a></li>
 +
                               
 +
                             
 +
</ul>
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</li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/Modeling"><span>MODELING</span></a>
 +
<ul>
 +
                                <li><a href="https://2015.igem.org/Team:SCUT/Overview">Overview</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/Biosensor">Biosensor</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/Bioeffector">Bioeffector</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/FutureWork">Future Work</a></li>
 +
 +
 +
</ul>
 +
</li>
 +
<li><a href="#"><span>RESULT</span></a>
 +
<ul>
 +
<li><a href="https://2015.igem.org/Team:SCUT/Parts">Parts</a></li>
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<li><a href="https://2015.igem.org/Team:SCUT/BasicParts">Basic Parts</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/Inthelab">In the lab</a></li>
 +
                                <li><a href="https://2015.igem.org/Team:SCUT/Description">Description</a></li>
 +
                                <li><a href="https://2015.igem.org/Team:SCUT/Measurement">Measurement</a></li>
 +
                                <li><a href="https://2015.igem.org/Team:SCUT/Collaboration">Collaboration</a></li>
 +
                               
 +
                               
 +
 +
</ul>
 +
</li>
 +
                        <li><a href="#"><span>PRACTICE</span></a>
 +
<ul>
 +
<li><a href="https://2015.igem.org/Team:SCUT/EconomicsandEthics">Economics and ethics</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/Outdoors">OUTDOORS</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/SynBiologyStudy">SYN-BIOLOGY STUDY</a></li>
 +
<li><a href="https://2015.igem.org/Team:SCUT/Visits">VISITS</a></li>
 +
 +
</ul>
 +
</li>
 +
 +
 +
 +
                        <li><a href="https://2015.igem.org/Team:SCUT/Safety"><span>SAFETY</span></a>
 +
 +
</li>
 +
                       
 +
                       
 +
</ul>
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</nav><!--end navigation-->
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<div class="clear"></div>
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<div class="title-wrapper">
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<div class="title-bg">
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<div class="title-content">
 +
<div class="three-fourth">
 +
<h2>Attributions</h2>
 +
</div>
 +
<div class="one-fourth column-last">
 +
<div class="portfolio-nav">
 +
<a href="#" class="prev-project">Previous Project</a>
 +
<a href="#" class="close-project">Close Project</a>
 +
<a href="#" class="next-project">Next Project</a>
 +
</div>
 +
</div>
 +
</div><!--end title-content-->
 +
</div>
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</div><!--end title-wrapper-->
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</section>
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<a title="Portfolio Image 1"><img src="images/portfolio/jj-royal.jpg" alt=" "></a>
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</div>
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<div class="slide">
 +
<a title="Portfolio Image 1"><img src="images/portfolio/jj-royal-2.jpg" alt=" "></a>
 +
</div>
 +
</div>
 +
<a href="#" class="prev"><img src="images/blog-arrow-prev.png" width="27" height="43" alt="Arrow Prev"></a>
 +
<a href="#" class="next"><img src="images/blog-arrow-next.png" width="27" height="43" alt="Arrow Next"></a>
 +
</div><!--end slides-->
 +
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<span class="masonry-post-meta">
 +
<a href="#">Branding</a> / <a href="#">Package Design</a>
 +
</span>
 +
<div class="clear"></div>
 +
 +
 +
<p class=MsoNormal><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:
 +
Arial;color:black;background:white'>In the iGEM competition, teams specify,
 +
design, build, and test simple biological systems made from standard,
 +
interchangeable biological parts.<span class=apple-converted-space>&nbsp;</span>Most
 +
BioBrick parts have never been characterized. And it was important to make a
 +
characterization for parts, which people could use the parameter as the experimental
 +
basis. Protein expression was a key parameter for a promoter. So in this part,
 +
we aimed at measuring the fluorescence of GFP expression which was activated by
 +
our promoter, using a plate reader.<o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:22.0pt;mso-bidi-font-family:
 +
Arial;color:black;background:white'>Introduction<o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:
 +
Arial;color:black;background:white'>We chose a promoter that had never been
 +
characterized in the register M36247 as our improvement work. M36247 was a constitutive
 +
promoter </span><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:
 +
Arial;background:white'>at medium strength in E.coli</span><span lang=EN-US
 +
style='font-size:14.0pt'>. The construction were inserted I13504 as a back
 +
insert into the promoter. <o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal align=left style='margin-bottom:3.6pt;text-align:left;
 +
line-height:14.3pt;mso-pagination:widow-orphan;mso-outline-level:3;background:
 +
white'><b><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:Arial;
 +
color:black;mso-font-kerning:0pt'>Strains:<o:p></o:p></span></b></p>
 +
 
 +
<p class=MsoNormal align=left style='margin-top:4.8pt;margin-right:0cm;
 +
margin-bottom:6.0pt;margin-left:0cm;text-align:left;line-height:14.3pt;
 +
mso-pagination:widow-orphan;background:white'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-family:Arial;color:black;mso-font-kerning:
 +
0pt'>The system should be measured in the strain of E.coli BL21.<o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal align=left style='margin-bottom:3.6pt;text-align:left;
 +
line-height:14.3pt;mso-pagination:widow-orphan;mso-outline-level:3;background:
 +
white'><b><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:Arial;
 +
color:black;mso-font-kerning:0pt'>Plasmid:<o:p></o:p></span></b></p>
 +
 
 +
<p class=MsoNormal align=left style='margin-top:4.8pt;margin-right:0cm;
 +
margin-bottom:6.0pt;margin-left:0cm;text-align:left;line-height:14.3pt;
 +
mso-pagination:widow-orphan;background:white'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-family:Arial;color:black;mso-font-kerning:
 +
0pt'>The Biobrick parts measured must be supplied in the plasmid pSB1C3.<o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal align=left style='margin-bottom:3.6pt;text-align:left;
 +
line-height:14.3pt;mso-pagination:widow-orphan;mso-outline-level:3;background:
 +
white'><b><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:Arial;
 +
color:black;mso-font-kerning:0pt'>Reporter:<o:p></o:p></span></b></p>
 +
 
 +
<p class=MsoNormal align=left style='margin-top:4.8pt;margin-right:0cm;
 +
margin-bottom:6.0pt;margin-left:0cm;text-align:left;line-height:14.3pt;
 +
mso-pagination:widow-orphan;background:white'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-family:Arial;color:black;mso-font-kerning:
 +
0pt'>The Part BBa_I13504 is chosen as the reporter of our reporter.<o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal align=left style='margin-top:4.8pt;margin-right:0cm;
 +
margin-bottom:6.0pt;margin-left:0cm;text-align:left;line-height:14.3pt;
 +
mso-pagination:widow-orphan;background:white'><b style='mso-bidi-font-weight:
 +
normal'><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:Roboto;
 +
background:white;mso-highlight:white;mso-font-kerning:0pt'>Equipment</span></b><b
 +
style='mso-bidi-font-weight:normal'><span lang=EN-US style='font-size:14.0pt;
 +
mso-bidi-font-family:Roboto;mso-font-kerning:0pt'>:<o:p></o:p></span></b></p>
 +
 
 +
<p class=MsoNormal align=left style='margin-top:4.8pt;margin-right:0cm;
 +
margin-bottom:6.0pt;margin-left:0cm;text-align:left;line-height:14.3pt;
 +
mso-pagination:widow-orphan;background:white'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-family:Roboto;color:black;background:
 +
white;mso-highlight:white;mso-font-kerning:0pt'>Infinite M200 with the software
 +
Magellan 6.5</span><b style='mso-bidi-font-weight:normal'><span lang=EN-US
 +
style='font-size:14.0pt;mso-bidi-font-family:Arial;mso-font-kerning:0pt'><o:p></o:p></span></b></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:22.0pt'>Protocol<o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:21.0pt;text-indent:-21.0pt;mso-list:l0 level1 lfo1'><![if !supportLists]><span
 +
lang=EN-US style='font-size:14.0pt;font-family:Wingdings;mso-fareast-font-family:
 +
Wingdings;mso-bidi-font-family:Wingdings;color:black'><span style='mso-list:
 +
Ignore'>l<span style='font:7.0pt "Times New Roman"'>&nbsp; </span></span></span><![endif]><span
 +
lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:Arial;color:black;
 +
background:white'>Construction <o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:
 +
Arial;color:black;background:white'>We got the promoter by the way of overlap
 +
PCR. After sequencing, </span><span lang=EN-US style='font-size:14.0pt'>the
 +
construction were inserted I13504 as a back insert into the promoter.</span><span
 +
lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:Arial;color:black;
 +
background:white'> <o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:21.0pt;text-indent:-21.0pt;mso-list:l0 level1 lfo1'><![if !supportLists]><span
 +
lang=EN-US style='font-size:14.0pt;font-family:Wingdings;mso-fareast-font-family:
 +
Wingdings;mso-bidi-font-family:Wingdings;color:black'><span style='mso-list:
 +
Ignore'>l<span style='font:7.0pt "Times New Roman"'>&nbsp; </span></span></span><![endif]><span
 +
lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:Arial;color:black;
 +
background:white'>Growing and measuring<o:p></o:p></span></p>
 +
 
 +
<p style='margin-top:4.8pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
 +
0cm;line-height:14.65pt;background:white'><span lang=EN-US style='font-size:
 +
14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:Arial'>1. Streaked
 +
a plate of the strain which contained M36247 listed in pSB1C3 .<o:p></o:p></span></p>
 +
 
 +
<p style='margin-top:4.8pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
 +
0cm;line-height:14.65pt;background:white'><span lang=EN-US style='font-size:
 +
14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:Arial'>2.
 +
Inoculated three 10ml cultures of supplemented M9 Medium and antibiotic (chloramphenicol
 +
</span><span lang=EN-US style='font-size:14.0pt;font-family:"Calibri",sans-serif;
 +
mso-bidi-font-family:Roboto;background:white;mso-highlight:white'>25</span><span
 +
lang=EN-US style='font-size:14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:
 +
Arial;letter-spacing:-.55pt;background:white;mso-highlight:white'>μ</span><span
 +
lang=EN-US style='font-size:14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:
 +
Arial'>g/ml) with single colony from the plate.<o:p></o:p></span></p>
 +
 
 +
<p style='margin-top:4.8pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
 +
0cm;line-height:14.65pt;background:white'><span lang=EN-US style='font-size:
 +
14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:Arial'>3. Cultures
 +
were grown in</span><span lang=EN-US style='font-size:9.5pt;font-family:Roboto;
 +
mso-bidi-font-family:Roboto;color:black;background:white;mso-highlight:white'> </span><span
 +
lang=EN-US style='font-size:14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:
 +
Roboto;color:black;background:white;mso-highlight:white'>50ml conical tube</span><span
 +
lang=EN-US style='font-size:14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:
 +
Arial'> for 16hours at 37</span><span lang=EN-US style='font-size:14.0pt;
 +
mso-ascii-font-family:Calibri'>℃</span><span lang=EN-US style='font-size:14.0pt;
 +
font-family:"Calibri",sans-serif;mso-bidi-font-family:Arial'> with shaking at 250rpm.<o:p></o:p></span></p>
 +
 
 +
<p style='margin-top:4.8pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
 +
0cm;line-height:14.65pt;background:white'><span lang=EN-US style='font-size:
 +
14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:Arial'>4. Cultures
 +
were diluted 1:100 into 3ml fresh medium and grown for 3hrs.<o:p></o:p></span></p>
 +
 
 +
<p style='margin-top:4.8pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
 +
0cm;line-height:14.65pt;background:white'><span lang=EN-US style='font-size:
 +
14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:Arial'>5. Measure
 +
the fluorescence (</span><span lang=EN-US style='font-size:14.0pt;font-family:
 +
"Calibri",sans-serif;mso-bidi-font-family:Roboto;color:black;background:white;
 +
mso-highlight:white'>Infinite M200 with the software Magellan 6.5</span><span
 +
lang=EN-US style='font-size:14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:
 +
Roboto;color:black'>,</span><span lang=EN-US style='font-size:14.0pt;
 +
font-family:"Calibri",sans-serif;mso-bidi-font-family:Arial'> 485 nm
 +
excitation, 528 nm emission) and absorbance (600nm) every 30 minutes in the
 +
next 4 hours.<o:p></o:p></span></p>
 +
 
 +
<p style='margin-top:4.8pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
 +
21.0pt;text-indent:-21.0pt;line-height:14.65pt;mso-list:l0 level1 lfo1;
 +
background:white'><![if !supportLists]><span lang=EN-US style='font-size:14.0pt;
 +
font-family:Wingdings;mso-fareast-font-family:Wingdings;mso-bidi-font-family:
 +
Wingdings'><span style='mso-list:Ignore'>l<span style='font:7.0pt "Times New Roman"'>&nbsp;
 +
</span></span></span><![endif]><span lang=EN-US style='font-size:14.0pt;
 +
font-family:"Calibri",sans-serif;mso-bidi-font-family:Arial'>Processing the
 +
data<o:p></o:p></span></p>
 +
 
 +
<p style='margin-top:4.8pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
 +
0cm;line-height:14.65pt;background:white'><span lang=EN-US style='font-size:
 +
14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:Arial;color:#282828'>Every
 +
device was measured thrice. The data was the arithmetic average of the three
 +
row data. Then they were subtracted the background controlling LB.<o:p></o:p></span></p>
 +
 
 +
<p style='margin-top:4.8pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
 +
21.0pt;text-indent:-21.0pt;line-height:14.65pt;mso-list:l0 level1 lfo1;
 +
background:white'><![if !supportLists]><span lang=EN-US style='font-size:14.0pt;
 +
font-family:Wingdings;mso-fareast-font-family:Wingdings;mso-bidi-font-family:
 +
Wingdings;color:#282828'><span style='mso-list:Ignore'>l<span style='font:7.0pt "Times New Roman"'>&nbsp;
 +
</span></span></span><![endif]><span lang=EN-US style='font-size:14.0pt;
 +
font-family:"Calibri",sans-serif;mso-bidi-font-family:Arial;color:#282828'>Positive
 +
and negative control<o:p></o:p></span></p>
 +
 
 +
<p style='margin-top:4.8pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
 +
0cm;line-height:14.65pt;background:white'><span lang=EN-US style='font-size:
 +
14.0pt;font-family:"Calibri",sans-serif;mso-bidi-font-family:Arial;color:#282828'>As
 +
our positive control, J23101 was medium strength promoter in constitutive
 +
family with close strength to our promoter, to exclude false negative results
 +
caused by the operation or low content. R0040 and BL21 without any plasmid were
 +
our negative control. R0040 was the part of ptet, which could regard as an
 +
empty plasmid to exclude false negative results caused by the operation or low
 +
content. Differed from the positive control, there was no back insert I13504. <o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:22.0pt;mso-bidi-font-family:
 +
Arial;color:black;background:white'>Result<o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:14.0pt;font-family:"Vijaya",sans-serif;
 +
color:black;background:white'>Figure 1: The expression of fluorescence was
 +
growing with the increasing of OD.<o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:14.0pt;font-family:"Vijaya",sans-serif;
 +
color:black;background:white;mso-no-proof:yes'>Figure 2</span><span
 +
style='font-size:14.0pt;font-family:宋体;mso-ascii-font-family:Vijaya;mso-hansi-font-family:
 +
Calibri;mso-bidi-font-family:Vijaya;color:black;background:white;mso-no-proof:
 +
yes'>：</span><span style='font-size:14.0pt;font-family:"Vijaya",sans-serif;
 +
color:black;background:white;mso-no-proof:yes'> <span lang=EN-US>OD of the
 +
three biological replicates were growing in the four hours. <o:p></o:p></span></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:14.0pt;font-family:"Vijaya",sans-serif;
 +
color:black;background:white;mso-no-proof:yes'>Figure 3: The fluorescence of
 +
three biological replicate of M36247+I13504 were growing in four hours.</span><span
 +
lang=EN-US style='font-size:14.0pt;font-family:"Vijaya",sans-serif;color:black;
 +
background:white'><o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:22.0pt;mso-bidi-font-family:
 +
Arial;color:black;background:white'>Discussion <o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:
 +
Arial;color:black;background:white'>It could be seen that M36247 was a constitutive
 +
promoter </span><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:
 +
Arial;background:white'>at medium strength, which could activated the
 +
expression of GFP without any inducer added. And compared with our positive
 +
control J23101, M36247 were slightly stronger. <o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:
 +
Arial;color:black;background:white;mso-no-proof:yes'><o:p>&nbsp;</o:p></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='font-size:14.0pt;mso-bidi-font-family:
 +
Arial;color:black;background:white'><o:p>&nbsp;</o:p></span></p>
 +
 
 +
 +
<div class="space"></div>
 +
 +
<h6>PROJECT PHOTOS</h6>
 +
 +
<div class="portfolio-gallery">
 +
<a title="Figure 1: The expression of fluorescence was growing with the increasing of OD" href="">
 +
 +
<img src="https://static.igem.org/mediawiki/parts/6/6c/2015SCUT_improvement1.png
 +
" alt=" " />
 +
</a>
 +
<a  title="Figure 2： OD of the three biological replicates were growing in the four hours" href="">
 +
 +
<img src="https://static.igem.org/mediawiki/parts/c/c3/2015SCUT_improvement2.png" alt=" " />
 +
</a>
 +
<a title="Figure 3: The fluorescence of three biological replicate of M36247+I13504 were growing in four hours" href="">
 +
 +
<img src="https://static.igem.org/mediawiki/parts/c/ca/2015SCUT_improvement3.png" alt=" " />
 +
</a>
 +
 +
 +
</div>
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<h3>About Us</h3>
 +
<p>In 2015, we SCUT teams won top ten innovative and entrepreneurial team set up by SCUT. In our team everyone owns basic biological knowledge and skills of the competition(iGEM). </p>
 +
<div class="footer-logo"></div>
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<div class="tweet"></div>
 +
</div>
 +
 +
<div class="percent-one-half column-last">
 +
<h3>Thanks</h3>
 +
<ul>
 +
<li>Zhang Zhenwu,Prof. Guo Shouqian,Dr. Li, Dr. Li Cheng,Dr. Wang Meng,Chen Kejie</li>
 +
<li>Guangzhou Municipal Environmental Protection Bureau<br/>
 +
</a></li>
 +
</ul>
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Revision as of 09:47, 17 September 2015

Team:SCUT/Discription

Attributions

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Branding / Package Design

In the iGEM competition, teams specify, design, build, and test simple biological systems made from standard, interchangeable biological parts. Most BioBrick parts have never been characterized. And it was important to make a characterization for parts, which people could use the parameter as the experimental basis. Protein expression was a key parameter for a promoter. So in this part, we aimed at measuring the fluorescence of GFP expression which was activated by our promoter, using a plate reader.

Introduction

We chose a promoter that had never been characterized in the register M36247 as our improvement work. M36247 was a constitutive promoter at medium strength in E.coli. The construction were inserted I13504 as a back insert into the promoter.

Strains:

The system should be measured in the strain of E.coli BL21.

Plasmid:

The Biobrick parts measured must be supplied in the plasmid pSB1C3.

Reporter:

The Part BBa_I13504 is chosen as the reporter of our reporter.

Equipment:

Infinite M200 with the software Magellan 6.5

Protocol

l  Construction

We got the promoter by the way of overlap PCR. After sequencing, the construction were inserted I13504 as a back insert into the promoter.

l  Growing and measuring

1. Streaked a plate of the strain which contained M36247 listed in pSB1C3 .

2. Inoculated three 10ml cultures of supplemented M9 Medium and antibiotic (chloramphenicol 25μg/ml) with single colony from the plate.

3. Cultures were grown in 50ml conical tube for 16hours at 37℃ with shaking at 250rpm.

4. Cultures were diluted 1:100 into 3ml fresh medium and grown for 3hrs.

5. Measure the fluorescence (Infinite M200 with the software Magellan 6.5, 485 nm excitation, 528 nm emission) and absorbance (600nm) every 30 minutes in the next 4 hours.

l  Processing the data

Every device was measured thrice. The data was the arithmetic average of the three row data. Then they were subtracted the background controlling LB.

l  Positive and negative control

As our positive control, J23101 was medium strength promoter in constitutive family with close strength to our promoter, to exclude false negative results caused by the operation or low content. R0040 and BL21 without any plasmid were our negative control. R0040 was the part of ptet, which could regard as an empty plasmid to exclude false negative results caused by the operation or low content. Differed from the positive control, there was no back insert I13504.

Result

Figure 1: The expression of fluorescence was growing with the increasing of OD.

Figure 2： OD of the three biological replicates were growing in the four hours.

Figure 3: The fluorescence of three biological replicate of M36247+I13504 were growing in four hours.

Discussion

It could be seen that M36247 was a constitutive promoter at medium strength, which could activated the expression of GFP without any inducer added. And compared with our positive control J23101, M36247 were slightly stronger.

 

 

PROJECT PHOTOS

About Us

In 2015, we SCUT teams won top ten innovative and entrepreneurial team set up by SCUT. In our team everyone owns basic biological knowledge and skills of the competition(iGEM).

We are

Thanks

  • Zhang Zhenwu,Prof. Guo Shouqian,Dr. Li, Dr. Li Cheng,Dr. Wang Meng,Chen Kejie
  • Guangzhou Municipal Environmental Protection Bureau

COPYRIGHT ©2015-SCUT