Difference between revisions of "Team:TU Dresden/Composite Part"
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<h1 class="box-heading">Short Description</h1> | <h1 class="box-heading">Short Description</h1> | ||
| − | <p>Peptidosomes in combination with Bacillus subtilis offer a perfect platform for enhanced protein overproduction by the means of efficient protein secretion and facilitated purification due to the peptidosomes’ pores. Naturally, B. subtilis is a strong secretion host and in order to take full advantage of this great potential it is necessary to evaluate all possible combinations of the B. subtilis’ secretion signal peptides and the proteins of interest. Therefore, we developed <b>the Evaluation Vector (EV)</b> which is an efficient genetic tool with a specially designed multiple cloning site to easily exchange translational fusions of the desired protein with the secretion signal peptides.</p> | + | <p>Peptidosomes in combination with <i>Bacillus subtilis</i> offer a perfect platform for enhanced protein overproduction by the means of efficient protein secretion and facilitated purification due to the peptidosomes’ pores. Naturally, <i>B. subtilis</i> is a strong secretion host and in order to take full advantage of this great potential it is necessary to evaluate all possible combinations of the <i>B. subtilis’</i> secretion signal peptides and the proteins of interest. Therefore, we developed <b>the Evaluation Vector (EV)</b> which is an efficient genetic tool with a specially designed multiple cloning site to easily exchange translational fusions of the desired protein with the secretion signal peptides.</p> |
</div> | </div> | ||
<div class="contentbox"> | <div class="contentbox"> | ||
<h1 class="box-heading">Background</h1> | <h1 class="box-heading">Background</h1> | ||
| − | <p>Tool development and their proper evaluation are core aspects of Synthetic Biology. In our project EncaBcillus one main idea was to establish Peptidosomes with encapsulated Bacteria as efficient protein overproduction platform. We took advantage of B. subtilis’ ability to efficiently secrete proteins into its environment in order to increase overall yields and to simplify the purification of the desired proteins.<br>Therefore, we developed a general expression evaluation vector (EV) with easily exchangeable units: I) allowing the replacement of the promoter (which drives the system) and II) a multiple cloning site enabling to work with translationally fused composite parts. In our case, a typical composite part consists of a signal peptide (for secretion in B. subtilis) and a protein of interest.</p> | + | <p>Tool development and their proper evaluation are core aspects of Synthetic Biology. In our project EncaBcillus one main idea was to establish Peptidosomes with encapsulated Bacteria as efficient protein overproduction platform. We took advantage of <i>B. subtilis’</i> ability to efficiently secrete proteins into its environment in order to increase overall yields and to simplify the purification of the desired proteins.<br>Therefore, we developed a general expression evaluation vector (EV) with easily exchangeable units: I) allowing the replacement of the promoter (which drives the system) and II) a multiple cloning site enabling to work with translationally fused composite parts. In our case, a typical composite part consists of a signal peptide (for secretion in <i>B. subtilis</i>) and a protein of interest.</p> |
| + | <p>In summary, our EV was designed to fulfill the following distinct features: | ||
| + | <ul> | ||
| + | <li>Exchangeable promoter region</li> | ||
| + | <li>Insertion of transcriptional and translational expression units</li> | ||
| + | <li>Fulfilling the RFC10 and RFC25 BioBrick standard</li> | ||
| + | <li>Easy cloning and screening procedure in <i>Escherichia coli</i></li> | ||
| + | </ul> | ||
| + | </p> | ||
</div> | </div> | ||
<div class="contentbox"> | <div class="contentbox"> | ||
Revision as of 13:11, 24 October 2017
Short Description
Peptidosomes in combination with Bacillus subtilis offer a perfect platform for enhanced protein overproduction by the means of efficient protein secretion and facilitated purification due to the peptidosomes’ pores. Naturally, B. subtilis is a strong secretion host and in order to take full advantage of this great potential it is necessary to evaluate all possible combinations of the B. subtilis’ secretion signal peptides and the proteins of interest. Therefore, we developed the Evaluation Vector (EV) which is an efficient genetic tool with a specially designed multiple cloning site to easily exchange translational fusions of the desired protein with the secretion signal peptides.
Background
Tool development and their proper evaluation are core aspects of Synthetic Biology. In our project EncaBcillus one main idea was to establish Peptidosomes with encapsulated Bacteria as efficient protein overproduction platform. We took advantage of B. subtilis’ ability to efficiently secrete proteins into its environment in order to increase overall yields and to simplify the purification of the desired proteins.
Therefore, we developed a general expression evaluation vector (EV) with easily exchangeable units: I) allowing the replacement of the promoter (which drives the system) and II) a multiple cloning site enabling to work with translationally fused composite parts. In our case, a typical composite part consists of a signal peptide (for secretion in B. subtilis) and a protein of interest.
In summary, our EV was designed to fulfill the following distinct features:
- Exchangeable promoter region
- Insertion of transcriptional and translational expression units
- Fulfilling the RFC10 and RFC25 BioBrick standard
- Easy cloning and screening procedure in Escherichia coli
Design
Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.
Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.
Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.
Results
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