Difference between revisions of "Team:SKLMT-China/Composite Part"
| Line 2: | Line 2: | ||
| − | <section class="article-banner" style="background-image: url(https://static.igem.org/mediawiki/2018/ | + | <section class="article-banner" style="background-image: url(https://static.igem.org/mediawiki/2018/6/6e/T--SKLMT-China--parts-banner-2.jpg)"> |
| + | <div class="banner-content"> | ||
<h2 class="title">Composite Part</h2> | <h2 class="title">Composite Part</h2> | ||
| − | + | <p class="content">What is four revolution?</p> | |
</div> | </div> | ||
<div class="container"> | <div class="container"> | ||
| Line 22: | Line 23: | ||
</div> | </div> | ||
<ul class="dropdown-menu"> | <ul class="dropdown-menu"> | ||
| − | <li><a href="#s1" class="scrolly | + | <li><a href="#s1" class="scrolly">Overview</a></li> |
| − | <li><a href="#s2" class="scrolly- | + | <li><a href="#s2" class="scrolly">PdnaA-NicA2</a></li> |
| + | <li><a href="#s2" class="scrolly">Composite Parts</a></li> | ||
</ul> | </ul> | ||
| Line 34: | Line 36: | ||
<div class="container" id="section-2"> | <div class="container" id="section-2"> | ||
| − | <div class="paragraph shadow | + | <div class="paragraph shadow"> |
| − | <h2 class="title">Overview </h2> | + | <h2 class="title" id="s1">Overview </h2> |
<p>Our submitted parts include a promoter library of inner promoters with various strength built in <latin>Pseudomonas fluorescence</latin>-pf5. We also include well-characterized composite part. One includes our strongest promoter and nicotine oxidase NicA2, working well in enhancing nicotine degradation compared to original nicA2 gene. </p> | <p>Our submitted parts include a promoter library of inner promoters with various strength built in <latin>Pseudomonas fluorescence</latin>-pf5. We also include well-characterized composite part. One includes our strongest promoter and nicotine oxidase NicA2, working well in enhancing nicotine degradation compared to original nicA2 gene. </p> | ||
</div> | </div> | ||
| − | <div class="paragraph shadow | + | <div class="paragraph shadow"> |
| − | <h2 class="title">PdnaA-NicA2 </h2> | + | <h2 class="title" id="s2">PdnaA-NicA2 </h2> |
<p>We used SDS-PAGE, to demonstrate the expression of NicA2 (52.5 kDa). In the picture, we can see the clear expression of NicA2 in <i>P.</i>pf-5 containing plasmid with our promoter. It can be qualitatively known that our promoters can initiate transcription normally, and the nicotine-degrading gene cluster has also been successfully heterologously expressed.</p> | <p>We used SDS-PAGE, to demonstrate the expression of NicA2 (52.5 kDa). In the picture, we can see the clear expression of NicA2 in <i>P.</i>pf-5 containing plasmid with our promoter. It can be qualitatively known that our promoters can initiate transcription normally, and the nicotine-degrading gene cluster has also been successfully heterologously expressed.</p> | ||
| Line 49: | Line 51: | ||
</div> | </div> | ||
| − | <div class="paragraph shadow | + | <div class="paragraph shadow"> |
| − | <h2 class="title">Composite Parts</h2> | + | <h2 class="title" id="s3">Composite Parts</h2> |
<p></p> | <p></p> | ||
Revision as of 17:14, 17 October 2018
Overview
Our submitted parts include a promoter library of inner promoters with various strength built in
PdnaA-NicA2
We used SDS-PAGE, to demonstrate the expression of NicA2 (52.5 kDa). In the picture, we can see the clear expression of NicA2 in P.pf-5 containing plasmid with our promoter. It can be qualitatively known that our promoters can initiate transcription normally, and the nicotine-degrading gene cluster has also been successfully heterologously expressed.
As for transcription, we are going to use real-time PCR to compare the ability of initiating transcription of three promoters. In addition, we are comparing the degradation efficiency by HPLC. NicA2 in can convert nicotine to pseudo-oxidation in a whole-cell reaction, and the rest of the gene cluster will convert pseudo-oxidation to 2,5-DHP.
However, due to time limitation, we were unable to complete the last two experiments, but we will do a supplementary explanation in our presentation. Please stay tuned.
Composite Parts
| Biobrick | Type | Description | length |
| BBa_K2569031 | Composite | PdnaA -NicA2 | 1525 |
| BBa_K2569032 | Composite | PdnaA-mRFP | 782 |