Difference between revisions of "Team:DTU-Denmark/Project/Overview"
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<div class="col-md-12"> | <div class="col-md-12"> | ||
| − | + | <h3><br></h3> | |
| − | + | <h1>Overview</h1> | |
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| − | + | Scroll down for more<br> | |
| − | + | <a href="#Project" class="btn btn-circle btn-transparent page-scroll"> | |
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| − | + | <h1 id="Project">Project</h1> | |
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| − | + | <div class="col-md-12 well overview-section" style="height: 350px"> | |
| − | </h1> | + | <h1> |
| − | <p>Over the summer we worked with nonribosomal peptide synthetases (NRPSs). We developed a <em>B. subtilis</em> strain capable of oligo-mediated genome engineering and used this strain to alter a NRPS. We also investigated methods of screening for novel products with desired activities.</p> | + | Introduction |
| + | </h1> | ||
| + | <p>Over the summer we worked with nonribosomal peptide synthetases (NRPSs). We developed a <em>B. subtilis</em> strain capable of oligo-mediated genome engineering and used this strain to alter a NRPS. We also investigated methods of screening for novel products with desired activities.</p> | ||
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| − | </h1> | + | <h1> |
| − | <p | + | Background |
| + | </h1> | ||
| + | <p>Nonribosomal peptide synthases (NRPSs) are large multimodular enzymes that synthesize nonribosomal peptides, which are short bioactive peptides with a broad range of functions, including antibiotics, immunosuppressants and anticancer drugs.</p> | ||
<div class="overview-readmore"><a href="/Team:DTU-Denmark/Project/Background">Read more</a></div> | <div class="overview-readmore"><a href="/Team:DTU-Denmark/Project/Background">Read more</a></div> | ||
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| − | </h1> | + | <h1> |
| − | <p | + | MAGE subtilis |
| + | </h1> | ||
| + | <p>Multiplex automated genome engineering (MAGE) utilises cyclical recombination with short oligonucleotides in order to achieve a high allelic replacement efficiency and can be used to quickly generate cell populations with varying phenotypes. We introduced oligo-mediated genome engineering into <i>Bacillus subtilis </i>and used it to alter a nonribosomal peptide synthetase.</p> | ||
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<p><a href="/Team:DTU-Denmark/Project/MAGE">Read more</a></p> | <p><a href="/Team:DTU-Denmark/Project/MAGE">Read more</a></p> | ||
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| + | <div class="col-md-12 well overview-section" style="height: 350px"> | ||
| + | <h1> | ||
| + | Tyrocidine | ||
| + | </h1> | ||
| + | <p>Tyrocidine is a mixture of non-ribosomal peptides. It can only be used topically due to its toxicity. We sought to express the tyrocidine synthase cluster in <i>B. subtilis</i> to make novel derivatives with oligo-mediated recombineering.</p> | ||
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<p><a href="/Team:DTU-Denmark/Project/Tyrocidine">Read more</a></p> | <p><a href="/Team:DTU-Denmark/Project/Tyrocidine">Read more</a></p> | ||
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| + | </div> | ||
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| + | <div class="col-md-4"> | ||
| + | <div class="col-md-12 well overview-section" style="height: 350px"> | ||
| + | <h1> | ||
| + | Lab-on-a-disc | ||
| + | </h1> | ||
| + | <p>Lab-on-a-disc is a potential screening method for our MAGE NRPS products. After screening for antimicrobial activity, prospective MAGE edited NRPS products could be tested for different beneficial properties. One example is a quick screening method for cytotoxicity.</p> | ||
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<p><a href="/Team:DTU-Denmark/Project/Chip">Read more</a></p> | <p><a href="/Team:DTU-Denmark/Project/Chip">Read more</a></p> | ||
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| + | <div class="col-md-4"> | ||
| + | <div class="col-md-12 well overview-section" style="height: 350px"> | ||
| + | <h1> | ||
| + | Intein | ||
| + | </h1> | ||
| + | <p>When one method fails the Synthesizer team has to innovate! We explored an alternative approach for generating short, cyclized peptides with similar length to tyrocidine by using self-splicing proteins. These short, self-splicing proteins that have no function in the proteins they are a part of, besides catalyzing their own excision after translation, are called inteins. The splicing creates a peptide bond between the two adjacent amino acids next to the inteins and we hypothesized that it could be used to make our cyclic peptides.</p> | ||
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<p><a href="/Team:DTU-Denmark/Project/Intein">Read more</a></p> | <p><a href="/Team:DTU-Denmark/Project/Intein">Read more</a></p> | ||
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| + | NRP Detection | ||
| + | </h1> | ||
| + | <p>Separation and identification of our non-ribosomal peptide synthetase (NRPS) products was determined by ultra-high performance liquid chromatography with diode array detection coupled to quadrupole time-of-flight mass spectrometry (UHPLC-DAD-QTOFMS). NRPS variants were identified by changes in mass-to-charge ratio and column retention time. </p> | ||
| − | + | <p><a href="/Team:DTU-Denmark/Project/Detection">Read more</a></p> | |
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Revision as of 13:56, 9 November 2015
<div class="jumbotron hero" style="background-image: url(https://2015.igem.org<app.models.UploadedFile object at 0x7ff51422db90>)">
<a href="https://igem.org" target="_blank"> <img src="https://2015.igem.org<app.models.UploadedFile object at 0x7ff51421a810>"> </a>
Scroll down for more
<a href="#Project" class="btn btn-circle btn-transparent page-scroll"> </a>
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Project
Introduction
Over the summer we worked with nonribosomal peptide synthetases (NRPSs). We developed a B. subtilis strain capable of oligo-mediated genome engineering and used this strain to alter a NRPS. We also investigated methods of screening for novel products with desired activities.
Background
Nonribosomal peptide synthases (NRPSs) are large multimodular enzymes that synthesize nonribosomal peptides, which are short bioactive peptides with a broad range of functions, including antibiotics, immunosuppressants and anticancer drugs.
<a href="/Team:DTU-Denmark/Project/Background">Read more</a>
MAGE subtilis
Multiplex automated genome engineering (MAGE) utilises cyclical recombination with short oligonucleotides in order to achieve a high allelic replacement efficiency and can be used to quickly generate cell populations with varying phenotypes. We introduced oligo-mediated genome engineering into Bacillus subtilis and used it to alter a nonribosomal peptide synthetase.
<a href="/Team:DTU-Denmark/Project/MAGE">Read more</a>
Tyrocidine
Tyrocidine is a mixture of non-ribosomal peptides. It can only be used topically due to its toxicity. We sought to express the tyrocidine synthase cluster in B. subtilis to make novel derivatives with oligo-mediated recombineering.
<a href="/Team:DTU-Denmark/Project/Tyrocidine">Read more</a>
<img src="https://2015.igem.org<app.models.UploadedFile object at 0x7ff5141f0050>">
Lab-on-a-disc
Lab-on-a-disc is a potential screening method for our MAGE NRPS products. After screening for antimicrobial activity, prospective MAGE edited NRPS products could be tested for different beneficial properties. One example is a quick screening method for cytotoxicity.
<a href="/Team:DTU-Denmark/Project/Chip">Read more</a>
Intein
When one method fails the Synthesizer team has to innovate! We explored an alternative approach for generating short, cyclized peptides with similar length to tyrocidine by using self-splicing proteins. These short, self-splicing proteins that have no function in the proteins they are a part of, besides catalyzing their own excision after translation, are called inteins. The splicing creates a peptide bond between the two adjacent amino acids next to the inteins and we hypothesized that it could be used to make our cyclic peptides.
<a href="/Team:DTU-Denmark/Project/Intein">Read more</a>
NRP Detection
Separation and identification of our non-ribosomal peptide synthetase (NRPS) products was determined by ultra-high performance liquid chromatography with diode array detection coupled to quadrupole time-of-flight mass spectrometry (UHPLC-DAD-QTOFMS). NRPS variants were identified by changes in mass-to-charge ratio and column retention time.
<a href="/Team:DTU-Denmark/Project/Detection">Read more</a>
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