Difference between revisions of "Team:DTU-Denmark/Proof"
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</blockquote> | </blockquote> | ||
| − | <p> | + | <p><i> This page is intended to give brief overview of how we fulfilled gold medal requirement #3. For much more information please visit <a href="https://2016.igem.org/Team:DTU-Denmark/molecular_toolbox">(Molecular Tools)</a>. </i></p> |
| − | + | ||
| − | </p> | + | <p>Yarrowia Lipolytica has a great potential to be a very versatile cell factory due to its ability to grow on a wide range of substrates. However, working with this unconventional yeast is troublesome due to the lack of molecular tools that can be used for genetic engineering. </p> |
| + | |||
| + | <p>We wanted to open up for the possibility of using Y. lipolytica in the future to produce any desired product while growing on any kind of substrate. Our aim was to develop a plasmid able to replicate in E. coli for easy cloning and propagation. The plasmid should also be compatible with Y. lipolytica along with being compatible with the BioBrick standard.</p> | ||
| + | |||
| + | <p>First step was to design the plasmid. We designed a plasmid (pSB1A8YL) based on the high copy plasmid Puc19 for replication in E.coli and pSL16-CEN1-1(227) for replication in Y. lipolytica. </p> | ||
| + | |||
| + | <figure class="figure"> | ||
| + | <img id="img2" class="enlarge img-responsive figure-img" src="https://static.igem.org/mediawiki/2016/thumb/f/f0/T--DTU-Denmark--pSB1A8YL_Sequence_map.png/603px-T--DTU-Denmark--pSB1A8YL_Sequence_map.png" alt="DESCRIPTION"> | ||
| + | <figcaption class="figure-caption"><strong>Figure 2:</strong> Sequence map of pSB1A8YL. The colored blocks represents the following: Orange: pUC19 part, Blue modified pSL16-CEN1-1(227) part, pink: BioBrick prefix, purple: BioBrick suffix, red: terminator, green selection markers, grey: origin of replication. The full annotated sequence can be found <a href="http://parts.igem.org/Part:BBa_K2117009">HERE</a>. </figcaption> | ||
| + | </figure> | ||
| + | |||
| + | <p>Our next step was to prove that our plasmid was compatible with the BioBrick standard. | ||
| + | We made three BioBricks by combining BioBricks already in the registry: the Anderson promoter (BBa_K880005) and paired this with the chromoproteins: amilCP (BBa_K592009), amilGFP (BBa_K592010) or mRFP(E1010). | ||
| + | We assembles these BioBricks in our plasmid and transformed them into chemically competent DH5alpha cells.</p> | ||
| + | |||
| + | <figure class="figure"> | ||
| + | <img id="img6" class="enlarge img-responsive figure-img" src="https://static.igem.org/mediawiki/2016/thumb/c/c7/T--DTU-Denmark--Cloning_flow1.png/800px-T--DTU-Denmark--Cloning_flow1.png" alt="DESCRIPTION"> | ||
| + | <figcaption class="figure-caption"><strong>Figure 6:</strong> Cloning flow of the test of pSB1A8YL in <i>E. coli</i>. The expression of the chromoproteins should yield a color ouput detectable by visual inspection.</figcaption> | ||
| + | </figure> | ||
| + | |||
| + | <p>The coloured colonies show that we were able to assemble a BioBrick device in our own designed plasmid, transform it into E. coli and express a chromoprotein proving that the BioBrick devices work. </p> | ||
| + | |||
| + | |||
| + | |||
</div> <!-- /overview--> | </div> <!-- /overview--> | ||
Revision as of 20:33, 18 October 2016
Title
leader under the title, short introduction. Ubique moderatius efficiantur eum et, dico oporteat recusabo ius cu, pro id modus sadipscing. Maluisset patrioque eum ad, mel eius doctus accommodare eu, minimum deleniti repudiandae mel ea. Noster nostrud diceret sea no. Eos an nullam molestiae signiferumque, vel ne laudem ignota oblique. Duo te luptatum percipitur signiferumque, at dicunt iriure dolorem his.
Section 1
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Someone famous in Source Title
This page is intended to give brief overview of how we fulfilled gold medal requirement #3. For much more information please visit (Molecular Tools).
Yarrowia Lipolytica has a great potential to be a very versatile cell factory due to its ability to grow on a wide range of substrates. However, working with this unconventional yeast is troublesome due to the lack of molecular tools that can be used for genetic engineering.
We wanted to open up for the possibility of using Y. lipolytica in the future to produce any desired product while growing on any kind of substrate. Our aim was to develop a plasmid able to replicate in E. coli for easy cloning and propagation. The plasmid should also be compatible with Y. lipolytica along with being compatible with the BioBrick standard.
First step was to design the plasmid. We designed a plasmid (pSB1A8YL) based on the high copy plasmid Puc19 for replication in E.coli and pSL16-CEN1-1(227) for replication in Y. lipolytica.
Our next step was to prove that our plasmid was compatible with the BioBrick standard. We made three BioBricks by combining BioBricks already in the registry: the Anderson promoter (BBa_K880005) and paired this with the chromoproteins: amilCP (BBa_K592009), amilGFP (BBa_K592010) or mRFP(E1010). We assembles these BioBricks in our plasmid and transformed them into chemically competent DH5alpha cells.
The coloured colonies show that we were able to assemble a BioBrick device in our own designed plasmid, transform it into E. coli and express a chromoprotein proving that the BioBrick devices work.
Section 2
Has ut facer debitis, quo eu agam purto. In eum justo aeterno. Sea ut atqui efficiantur, mandamus deseruisse at est, erat natum cum eu. Quot numquam in vel. Salutatus euripidis moderatius qui ex, eu tempor volumus vituperatoribus has, ius ea ullum facer corrumpit.
Section 2.1
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Section 2.2
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Section 2.3
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Section 3
Has ut facer debitis, quo eu agam purto. In eum justo aeterno. Sea ut atqui efficiantur, mandamus deseruisse at est, erat natum cum eu. Quot numquam in vel. Salutatus euripidis moderatius qui ex, eu tempor volumus vituperatoribus has, ius ea ullum facer corrumpit.
Section 4
Has ut facer debitis, quo eu agam purto. In eum justo aeterno. Sea ut atqui efficiantur, mandamus deseruisse at est, erat natum cum eu. Quot numquam in vel. Salutatus euripidis moderatius qui ex, eu tempor volumus vituperatoribus has, ius ea ullum facer corrumpit.
Section 5
Has ut facer debitis, quo eu agam purto. In eum justo aeterno. Sea ut atqui efficiantur, mandamus deseruisse at est, erat natum cum eu. Quot numquam in vel. Salutatus euripidis moderatius qui ex, eu tempor volumus vituperatoribus has, ius ea ullum facer corrumpit.
