Difference between revisions of "Team:NUDT CHINA/Improve"

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{{NUDT_CHINA}}
 
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<html>
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<head>
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                    <h1 class="main-title">Improve</span></h1>
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                </div>
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<div class="column full_size judges-will-not-evaluate">
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        <div class="content" style="width: 74%">
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<h3>★  ALERT! </h3>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2017.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2017.igem.org/Judging/Awards"> award listed above</a>. </p>
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2017.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
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<p class="content-subhead">BBa_I712019</p>
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<p>This summer, we wanted to construct a dual-luciferase miRNA target expression plasmid and perform a dual luciferase assay, to evaluate the microRNA (miRNA) binding ability of miRNA binding sequences.</p>
 +
<p>MiR-214 and its target sequence RNF8 were chosen in our project, unfortunately, the 3&rsquo; untranslated region of RNF8 is not compatible with BioBrick RFC[10] as multiple illegal sites were contained in it. This troubles us a lot and caused a lot of difficulties in our experiment.</p>
 +
<p>Thus, we decided to do something to improve the part BBa_I712019, to optimize its function in a dual-luciferase miRNA target expression system. Two restriction sites of BsmBI were added to the 3&rsquo; end of firefly luciferase, through which, the 3&rsquo; untranslated region of any gene can be inserted using Golden-Gate Assembly, with no worry about the multiple illegal sites contained in it.</p>
 +
<p>We believe that this improvement simplifies the construction of a dual-luciferase miRNA target expression plasmid and greatly facilitates the related miRNA research.</p>
 +
<center><img width="65%" src="https://static.igem.org/mediawiki/2017/4/40/T-NUDT_CHINA-Improve1.jpg" alt=""><p style="font: caption;text-indent: 0;"><strong>Figure 1. Schematic representation of the workflow of the plasmid construction.</strong> (a) The improvement of the part. (b) The restriction sites of BsmBI. (c) The construction of the plasmid.</p></center>
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<div class="column full_size">
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<script type="text/javascript">
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<h1>Improve</h1>
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<p>For teams seeking to improve upon a previous part or project, you should document all of your work on this page. Please remember to include all part measurement and characterization data on the part page on the Regisrty. Please include a link to your improved part on this page.</p>
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<h3>Gold Medal Criterion #2</h3>
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</script>
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<p><b>Standard Tracks:</b> Improve the function of an existing BioBrick Part. The original part must NOT be from your 2017 part number range. If you change the original part sequence, you must submit a new part. In addition, both the new and original part pages must reference each other. This working part must be different from the part documented in bronze #4 and silver #1.
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<b>Special Tracks:</b> Improve the function of an existing iGEM project (that your current team did not originally create) and display your achievement on your wiki.</p>
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Latest revision as of 03:12, 2 November 2017

HOME
TEAM
PROJECT
Description
Design
Experiments
Notebook
InterLab
Contribution
Model
Results
Demonstrate
Improve
Attributions
PARTS
Parts
Basic Parts
Composite Parts
Part Collection
SAFETY
HUMAN PRACTICES
Silver HP
Integrated and Gold
Public Engagement
AWARDS
Applied Design
Entrepreneurship
Hardware
Measurement
Model
Plant
Software
JUDGING FORM

Improve

BBa_I712019

This summer, we wanted to construct a dual-luciferase miRNA target expression plasmid and perform a dual luciferase assay, to evaluate the microRNA (miRNA) binding ability of miRNA binding sequences.

MiR-214 and its target sequence RNF8 were chosen in our project, unfortunately, the 3’ untranslated region of RNF8 is not compatible with BioBrick RFC[10] as multiple illegal sites were contained in it. This troubles us a lot and caused a lot of difficulties in our experiment.

Thus, we decided to do something to improve the part BBa_I712019, to optimize its function in a dual-luciferase miRNA target expression system. Two restriction sites of BsmBI were added to the 3’ end of firefly luciferase, through which, the 3’ untranslated region of any gene can be inserted using Golden-Gate Assembly, with no worry about the multiple illegal sites contained in it.

We believe that this improvement simplifies the construction of a dual-luciferase miRNA target expression plasmid and greatly facilitates the related miRNA research.

Figure 1. Schematic representation of the workflow of the plasmid construction. (a) The improvement of the part. (b) The restriction sites of BsmBI. (c) The construction of the plasmid.