Difference between revisions of "Part:BBa K5299008"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K5299008 short</partinfo> | <partinfo>BBa_K5299008 short</partinfo> | ||
| + | <span class='h3bb'><p>Sequence and Features</p></span> | ||
| + | <partinfo>BBa_K5299008 SequenceAndFeatures</partinfo> | ||
| + | <partinfo>BBa_K5299008 parameters</partinfo> | ||
Synthetic promoter identified by Zobel et all (2015) | Synthetic promoter identified by Zobel et all (2015) | ||
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| + | <h1 style="color:#3c6307;"><b>Introduction</b></h1> | ||
| + | Synthetic biology constitutes an effort towards making biology easy to engineer [1]. This means that basic principles of engineering find their place in biological systems; | ||
| + | We treat biomers as spare, interchangeable parts, the same way that we would approach the construction of any mechanical device. This necessitates the standardization of parts, in order to establish objectivity regarding their effectiveness and to promote the acceleration of knowledge [2,3]. For our project, to identify the optimal components and regulatory mechanisms for our system, we employed the Design-Build-Test-Learn cycle, allowing us to manipulate the expression of our constructs throughout different phases of the bacterial life cycle. To minimize cellular stress, we strategically divided our system into two phases: the exponential phase and the stationary phase. During the exponential phase, we expressed T7 polymerase and dsRNA molecules. In our pursuit of an autoinducible promoter active during the exponential phase, we explored the work of Zobel et al. and tested three of their synthetic promoters. Through our focused examination of their autoinducible properties, we determined that BG37 emerged as the optimal choice for regulating our system and we decided to further characterize BG37 by assessing its performance across various bacterial chassis, employing different plasmid backbones and carbon sources. By thoroughly characterizing this new basic part, we believe it will serve as a valuable tool for future teams aiming for orthogonal expression during the exponential phase. | ||
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| + | <html> | ||
| + | <center> | ||
| + | <figure> | ||
| + | <img src='https://static.igem.wiki/teams/5299/registry-nef/wet-lab.png' width='900px' height='800px' | ||
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| + | <figcaption><center><b><small><i>Figure 1: Production of T7 polymerase, regulated by BG37 promoter.</i></small></b></center></figcaption> | ||
| + | </figure></center></html> | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
| + | <h1 style="color:#3c6307;"><b>The origin of BG37</b></h1> | ||
| + | Zobel et al. identified the BG synthetic promoters by systematically analyzing promoter activities in E. coli and Pseudomonas strains, particularly P. aeruginosa and P. putida. They found that the consensus sequences, especially the −10 and −35 regions, closely resembled those of sigma-70 promoters in E. coli, which suggested similar transcriptional mechanisms across these species. Using an initial plasmid-based selection in E. coli PIR2 cells, they efficiently screened for effective synthetic promoters, confirming their comparable activity in both E. coli and Pseudomonas [4]. | ||
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| − | + | <center> | |
| − | < | + | <figure> |
| + | <img src='https://static.igem.wiki/teams/5299/registry-nef/images-medium-sb-2015-00058h-0008.gif' width='900px' height='900px' | ||
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| + | <figcaption><center><b><small><i>Figure 2: Figure 3: Sequence of BG Synthetic Promoters.</i></small></b></center></figcaption> | ||
| + | </figure></center></html> | ||
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| + | <span class='h3bb'>Sequence and Features</span> | ||
| + | <partinfo>BBa_K5299008 SequenceAndFeatures</partinfo> | ||
Revision as of 15:36, 26 September 2024
BG37: Autoinducible promoter in exponential phase
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Synthetic promoter identified by Zobel et all (2015)
Introduction
Synthetic biology constitutes an effort towards making biology easy to engineer [1]. This means that basic principles of engineering find their place in biological systems; We treat biomers as spare, interchangeable parts, the same way that we would approach the construction of any mechanical device. This necessitates the standardization of parts, in order to establish objectivity regarding their effectiveness and to promote the acceleration of knowledge [2,3]. For our project, to identify the optimal components and regulatory mechanisms for our system, we employed the Design-Build-Test-Learn cycle, allowing us to manipulate the expression of our constructs throughout different phases of the bacterial life cycle. To minimize cellular stress, we strategically divided our system into two phases: the exponential phase and the stationary phase. During the exponential phase, we expressed T7 polymerase and dsRNA molecules. In our pursuit of an autoinducible promoter active during the exponential phase, we explored the work of Zobel et al. and tested three of their synthetic promoters. Through our focused examination of their autoinducible properties, we determined that BG37 emerged as the optimal choice for regulating our system and we decided to further characterize BG37 by assessing its performance across various bacterial chassis, employing different plasmid backbones and carbon sources. By thoroughly characterizing this new basic part, we believe it will serve as a valuable tool for future teams aiming for orthogonal expression during the exponential phase.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]