Team:York/Cake

From 2014.igem.org

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                                                         <li class="active"><a href="https://2014.igem.org/Team:York/Constructs">Constructs</a></li>
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<h1>Constructs</h1>
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<h1>Team members</h1>
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<img class="img-responsive constructs" src="https://static.igem.org/mediawiki/2014/a/ae/York_Constructs.jpg">
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<p>In just 50 years, the University has become one of the top in the UK. We think this is because of its passion and dedication towards each of its subjects; something we share! Our team is made up of 20 undergraduates from a range of disciplines, as well as our two supervisors and advisor.</p>
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<br><br>
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<h2>Undergraduates</h2>
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<ul class="nav nav-tabs" role="tablist" id="myTab">
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    <li class="active"><a href="#one" role="tab" data-toggle="tab">pYodA</a></li>
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    <li><a href="#two" role="tab" data-toggle="tab">NRAMP</a></li>
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    <li><a href="#three" role="tab" data-toggle="tab">CysP</a></li>
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    <li><a href="#four" role="tab" data-toggle="tab">SpPCS</a></li>
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    <li><a href="#five" role="tab" data-toggle="tab">Gsh1*</a></li>
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<img src="https://static.igem.org/mediawiki/2014/2/25/York_RW.JPG" class="headshot img-responsive">
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<h4>Robyn Whiting</h4> <p><i>BSc Genetics First year</i></p>
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<div class="col-md-4">
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<img src="https://static.igem.org/mediawiki/2014/f/f6/York_SA.png" class="headshot img-responsive">
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<h4>Sarah Andrews</h4> <p><i>BSc Molecular Cell Biology First year</i></p>
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<h2>pYodA</h2>
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<img src="https://static.igem.org/mediawiki/2014/e/e4/York_NP.png" class="headshot img-responsive">
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<b>Name:</b> pYodA (ZinTp) <br>
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<h4>Nikola Panayotov</h4> <p><i>BSc Biochemistry Second year</i></p>
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<b>Organism:</b> <i>Escherichia Coli</i> <br>
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</div>
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<b>Normal Function:</b> pYodA is a cadmium-induced promoter that activates the yodA(zinT) gene, leading to the production of ZinT metal-binding protein.<br>
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<b>Aim:</b> We are using pYodA in an alternative way; to regulate the expression of the CysP gene (sulfate transporter) and the NRAMP gene (Cadmium transporter). Due to using pYodA, the expression of these two genes will be regulated by the concentration of Cadmium in the extracellular environment. The over-production of Cysteine will only occur when the concentration of Cadmium reaches the sensitivity threshold of pYodA. Due to cysteine production being both energetically demanding and toxic at high concentrations, we do not want these genes to be expressed constitutively. <br>
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</div>
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<b>Characterisation:</b> In order to characterise pYodA, we will couple it with a GFP gene. This will allow us to calculate the sensitivity threshold of pYodA and measure the amount of cadmium that can be chelated at various concentrations. <br>
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<div class="row team">
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<b>Literature:</b>
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<ol>
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<div class="col-md-4">
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<li>http://mic.sgmjournals.org/content/148/12/3801.long</li>
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<img src="https://static.igem.org/mediawiki/2014/e/e1/York_IG.JPG" class="headshot img-responsive">
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<li>http://www.uniprot.org/uniprot/F4VWH2</li>
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<h4>Ivan Gyulev</h4> <p><i>BSc Genetics Third year</i></p>
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<li>http://sbkb.org/uid/F4VWH2/uniprot#structures</li>
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</div>
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<li>http://www.jbc.org/content/278/44/43728</li>
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<div class="col-md-4">
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<li>http://biocyc.org/ECOLI/NEW-IMAGE?type=GENE&object=G7061</li>
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<img src="https://static.igem.org/mediawiki/2014/2/2b/York_HE.JPG" class="headshot img-responsive">
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</ol>
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<h4>Hanna Esser</h4> <p><i>BSc Biochemistry First year</i></p>
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</p>
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</div>
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<div class="col-md-4">
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<img src="https://static.igem.org/mediawiki/2014/6/69/York_RH.JPG" class="headshot img-responsive">
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<h4>Ruth Haley</h4> <p><i>BSc Biochemistry First year</i></p>
 +
</div>
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 +
</div>
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<img src="https://static.igem.org/mediawiki/2014/6/6a/York_LD.jpg" class="headshot img-responsive">
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<h4>Lindsey Dalzell</h4> <p><i>BSc Biology First year</i></p>
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</div>
 +
<div class="col-md-4">
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<img src="https://static.igem.org/mediawiki/2014/6/68/York_ED.png" class="headshot img-responsive">
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<h4>Ellie Davis</h4> <p><i>BSc Biology First year</i></p>
 +
</div>
 +
<div class="col-md-4">
 +
<img src="https://static.igem.org/mediawiki/2014/4/4a/York_JT.JPG" class="headshot img-responsive">
 +
<h4>Joseph Tresise</h4> <p><i>MChem Chemistry Third year</i></p>
 +
</div>
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</div>
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<div class="row team">
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                <div class="col-md-4">
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<img src="https://static.igem.org/mediawiki/2014/c/cf/York_RCdP.JPG" class="headshot img-responsive">
 +
<h4>Ricardo Cañavate del Pino</h4> <p><i>BSc Biochemistry Second year</i></p>
 +
</div>
 +
<div class="col-md-4">
 +
<img src="https://static.igem.org/mediawiki/2014/2/28/York_TM.jpg" class="headshot img-responsive">
 +
<h4>Teodora Manea</h4> <p><i>BSc Biochemistry First year</i></p>
 +
</div>
 +
<div class="col-md-4">
 +
<img src="https://static.igem.org/mediawiki/2014/d/db/York_CW.JPG" class="headshot img-responsive">
 +
<h4>Cauã Westmann </h4> <p><i>BSc Biology Third year</i></p>
 +
</div>
 +
</div>
 +
 
 +
<div class="row team">
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<div class="col-md-4">
 +
<img src="https://static.igem.org/mediawiki/2014/e/e2/York_MA.jpg" class="headshot img-responsive">
 +
<h4>Marieta Avramov</h4> <p><i>BSc Molecular Cell Biology First year</i></p>
 +
</div>
 +
<div class="col-md-4">
 +
<img src="https://static.igem.org/mediawiki/2014/3/39/York_AM.JPG" class="headshot img-responsive">
 +
<h4>Auriane Muse</h4> <p><i>BSc Biology Second year</i></p>
 +
</div>
</div>
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<h2>NRAMP</h2>
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<br>
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<b>Gene:</b> mntH<br>
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<ul>
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<b>Organism:</b> <i>Escherichia Coli</i><br>
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<li><p>Vishnu Sunil: <i>BSc Environmental Science Second year</i> </p></li>
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<b>Protein:</b> NRAMP<br>
+
<li><p>Lucy Dye: <i>BSc Biology First year</i> </p></li>
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<b>Function:</b> NRAMP is a membrane divalent metal-ion transporter. In addition to iron and manganese, it also transports Cd ions into the cell.<br>
+
<li><p>Maria Agapiou: <i>BSc Biology First year</i> </p></li>
-
<b>Aim:</b> We want to use the endogenous transporter NRAMP to take up the cadmium from the environment. The cadmium will then activate pYoda and set off the phytochelatin synthesis process.<br>
+
<li><p>Tsvetelina Tsekova: <i>BSc Genetics Second year</i> </p></li>
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<b>Expression:</b> We can run assays measuring the concentration of cadmium in the environment. If NRAMP is active, the concentration of cadmium should decrease.<br></p></div>
+
<li><p>Arushi Aneja: <i>MEng Computer Science with Artificial Intelligence</i> </p></li>
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<div class="tab-pane" id="three" class="collapse">
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</ul>
 +
</div></div>
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<h2>CysP</h2>
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 +
<h2>Acknowledgements</h2>
<p>
<p>
-
<b>Gene:</b> cysP (AKA YlnA) <br>
+
<b>Dr Gavin Thomas, Dr James Chong, Dr Thorunn Helgason, Dr Leo Caves</b> and <b>Professor Maggie Smith</b> for providing their input and advice as Senior Lecturers. Our team is indebted to them for the guidance they gave regarding the feasibility of our project at the start and supervising our progress throughout.
-
<b>Organism:</b> <i>Bacillus Subtilis</i> <br>
+
<b>Rosanna Hennessy</b> for helping us design our constructs and ensuring a trouble-free synthesis.
-
<b>Protein:</b> CysP <br>
+
<b>Michael Schultz, Daniel Ungar</b> and <b>Erica Kimtz</b> for attending our Sponsors event and giving input and feedback about our presentation and project.
-
<b>Function:</b> This gene encodes for CysP, a sulfate/thiosulfate ABC transporter found in the periplasmic space. <br>
+
The lab technicians, <b>Jen, Sam</b> and <b>Nikki</b> who kept our lab supplied and organised, while helping with any technical problems that arose over the summer.  
-
<b>Aim:</b> We want to overexpress this gene in E.coli. This would lead to increased sulfate uptake, which could be used to produce cysteine and eventually phytochelatins that would bind cadmium. <br>
+
<b>Dr Erica Kintz</b> and <b>Dr Marja van der Wouden</b> for providing us with some E. coli mutants and with 96 well plates for our characterisation.  
-
<b>Expression:</b> We could run assays measuring the concentration of sulfate in the environment containing our bacteria. <br>
+
<b>Dr Andrew Leech</b> for giving us advice on the design of our characterisation experiments.
 +
<b>Phil Roberts</b> for providing design advice that created this wiki you are reading. <br> <br>
 +
The 2014 York iGEM Team would like to thank all of the above for their advice and guidance throughout this competition, we know it wouldn't have been possible without these people giving up their time to aid us! Thank you very much!
</p>
</p>
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<h2>CysE*</h2>
 
-
<p>
 
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<b>Original Gene:</b> cysE<br>
 
-
<b>Mutant Gene:</b> cysE*<br>
 
-
<b>Organism:</b> <i>Escherichia Coli</i> <br>
 
-
<b>Protein:</b> CysE(SAT)<br>
 
-
<b>Normal function:</b> To catalyse the acetylation of L-serine (the first step in cysteine biosynthesis)<br>
 
-
<b>Function:</b> Serine acetyltransferase (CysE) carries out the first step in cysteine biosynthesis; it  catalyses the acetylation of L-serine which generates O-acetyl-L-serine. Cysteine itself strongly inhibits the activity of serine acetyltransferase by binding to the serine-binding site. This inhibition depends on the protein's carboxy terminus, and has been localized to Met-256 specifically. Due to cysteine production being both energetically demanding and toxic at high concentrations, the cell does not want to produce cysteine constitutively.<br>
 
-
<b>Aim:</b> To over-produce cysteine.To fulfill this aim, we need to remove negative feedback from cysteine biosynthesis. To do this, we are using a mutant CysE gene (CysE*). Our mutant CysE gene will produce a protein that has a single amino acid substitution: Met-256 will be replaced by Ile by changing the corresponding AUG codon to AUC. This single amino-acid substitution will alter the three dimensional shape of the serine-binding site in our acetyltransferase. Changing the shape of the serine-binding site prevents cysteine from binding to it. Thus, our acetyltransferase will not be inhibited by cysteine. As a result, we will be able to over-produce cysteine in our cell.<br>
 
-
<b>Method:</b> We had the cysE* gene synthesised to produce the mutant CysE* protein.<br>
 
-
<b>Literature:</b>
 
-
<ol>
 
-
<li>http://aem.asm.org/content/66/10/4497.full</li>
 
-
<li>Denk D., Bock A. <i>J. Gen. Microbiol</i>, 1987</li>
 
-
</ol></p></div>
 
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<div class="tab-pane" id="five" class="collapse">
 
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<h2>Gsh1*</h2>
 
-
<p>
 
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<b>Original Gene:</b> gsh1/gshA<br>
 
-
<b>Organism:</b> <i>Saccharomyces cerevisiae</i><br>
 
-
<b>Protein:</b> Glutamate-Cysteine Ligase (previously known as gamma-glutamylcysteine synthetase)<br>
 
-
<b>Function:</b> Gamma glutamylcysteine synthetase catalyzes the first step in glutathione (GSH) biosynthesis;<br>
 
-
L-glutamate + L-cysteine + ATP -> gamma-glutamyl cysteine + ADP + Pi<br>
 
-
Expression is induced by oxidants, cadmium, and mercury. Protein abundance increases in response to DNA replication stress.<br>
 
-
<b>Aim:</b> We can use the overproduced cysteine to make gamma-glutamyl cysteine, which is the monomer that forms phytochelatins (n=10-20). In order to do this, we need glutamate-cysteine ligase to catalyse the reaction. We can overexpress GSH1* using the pYodA promoter. <br>
 
-
<b>Literature:</b><ol><li>http://biocyc.org/YEAST/NEW-IMAGE?type=GENE-IN-MAP-IN-PWY&object=YJL101C</li></ol></p></div>
 
-
 
-
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<h2>spPCS</h2>
 
-
<p>
 
-
<b>Gene:</b> spPCS<br>
 
-
<b>Organism:</b> <i>Schizosaccharomyces pombe</i><br>
 
-
<b>Function:</b> The phytochelatin synthase in S. pombe uses glutathione with a blocked thiol group  to synthesise phytochelatins.<br>
 
-
<b>Aim:</b> Overexpression of this gene, together with Gsh1*, in E.coli has been shown to increase phytochelatin production and lead to a 7.5-times-higher Cd accumulation. We want to use this gene to make our bacteria more efficient in  taking up cadmium.<br>
 
-
<b>Literature</b><ol><li>http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2075016/</li></ol></p></div>
 
-
 
</div>
</div>
</div>
</div>
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Revision as of 12:36, 16 October 2014

Team York 2014


Team members

In just 50 years, the University has become one of the top in the UK. We think this is because of its passion and dedication towards each of its subjects; something we share! Our team is made up of 20 undergraduates from a range of disciplines, as well as our two supervisors and advisor.

Undergraduates

Robyn Whiting

BSc Genetics First year

Sarah Andrews

BSc Molecular Cell Biology First year

Nikola Panayotov

BSc Biochemistry Second year

Ivan Gyulev

BSc Genetics Third year

Hanna Esser

BSc Biochemistry First year

Ruth Haley

BSc Biochemistry First year

Lindsey Dalzell

BSc Biology First year

Ellie Davis

BSc Biology First year

Joseph Tresise

MChem Chemistry Third year

Ricardo Cañavate del Pino

BSc Biochemistry Second year

Teodora Manea

BSc Biochemistry First year

Cauã Westmann

BSc Biology Third year

Marieta Avramov

BSc Molecular Cell Biology First year

Auriane Muse

BSc Biology Second year


  • Vishnu Sunil: BSc Environmental Science Second year

  • Lucy Dye: BSc Biology First year

  • Maria Agapiou: BSc Biology First year

  • Tsvetelina Tsekova: BSc Genetics Second year

  • Arushi Aneja: MEng Computer Science with Artificial Intelligence

Acknowledgements

Dr Gavin Thomas, Dr James Chong, Dr Thorunn Helgason, Dr Leo Caves and Professor Maggie Smith for providing their input and advice as Senior Lecturers. Our team is indebted to them for the guidance they gave regarding the feasibility of our project at the start and supervising our progress throughout. Rosanna Hennessy for helping us design our constructs and ensuring a trouble-free synthesis. Michael Schultz, Daniel Ungar and Erica Kimtz for attending our Sponsors event and giving input and feedback about our presentation and project. The lab technicians, Jen, Sam and Nikki who kept our lab supplied and organised, while helping with any technical problems that arose over the summer. Dr Erica Kintz and Dr Marja van der Wouden for providing us with some E. coli mutants and with 96 well plates for our characterisation. Dr Andrew Leech for giving us advice on the design of our characterisation experiments. Phil Roberts for providing design advice that created this wiki you are reading.

The 2014 York iGEM Team would like to thank all of the above for their advice and guidance throughout this competition, we know it wouldn't have been possible without these people giving up their time to aid us! Thank you very much!

Retrieved from "http://2014.igem.org/Team:York/Cake"