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Team:DTU-Denmark/SPL Section
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<h1>Introduction</h1> | <h1>Introduction</h1> | ||
| - | <p align="justify"> | + | <p align="justify"> Not only was the Synthetic Promoter Library (SPL) used as a method for attempting to characterize lambda’s <a href="https://2010.igem.org/Team:DTU-Denmark/AntiTermination_Section" target="_blank"> N-antiterminator</a> protein, but was also used as a proof of concept experiment for the <a href="http://openwetware.org/wiki/The_BioBricks_Foundation:RFC#BBF_RFC_63:_DTU_Synthetic_Promoter_Library_Standard" target="_blank"> BBF RFC 63 – DTU Synthetic Promoter Library Standard</a> where we decided to ligate it in front of <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_I13507" target="_blank"> BBa_I13507</a>. <br></p> |
<a name="Construction"></a><h1>Construction of BioBricks</h1> | <a name="Construction"></a><h1>Construction of BioBricks</h1> | ||
| + | <p align="justify"> The SPL + I13507 construct was ligated into the BioBrick plasmid backbone pSB3T5, this due to pSB3T5 being a low to medium copy number plasmid which contains the p15A replication of origin and a tetracycline resistance marker. The expression of RFP via an SPL would best be controlled and measured through a low copy number plasmid in case too high expression of RFP proved to be detrimental to the cell’s viability, therefore pSB3T5 was chosen as the backbone as it had the best range of copy number amongst the other <a href="http://partsregistry.org/Plasmid_backbones/Assembly" target="_blank"> backbones</a> within the parts-registry. | ||
| + | The pSB2K3 backbone could have been used as well, though pSB3T5 gave us more flexibility with the construct as additional regulatory elements were not required such as the case with pSB2K3. <br></p> | ||
<a name="Characterization"></a><h1>Characterization</h1> | <a name="Characterization"></a><h1>Characterization</h1> | ||
<a name="Characterization_Strategy"></a><h3>Strategy</h3> | <a name="Characterization_Strategy"></a><h3>Strategy</h3> | ||
Revision as of 13:40, 27 October 2010
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Introduction Not only was the Synthetic Promoter Library (SPL) used as a method for attempting to characterize lambda’s N-antiterminator protein, but was also used as a proof of concept experiment for the BBF RFC 63 – DTU Synthetic Promoter Library Standard where we decided to ligate it in front of BBa_I13507. Construction of BioBricks The SPL + I13507 construct was ligated into the BioBrick plasmid backbone pSB3T5, this due to pSB3T5 being a low to medium copy number plasmid which contains the p15A replication of origin and a tetracycline resistance marker. The expression of RFP via an SPL would best be controlled and measured through a low copy number plasmid in case too high expression of RFP proved to be detrimental to the cell’s viability, therefore pSB3T5 was chosen as the backbone as it had the best range of copy number amongst the other backbones within the parts-registry.
The pSB2K3 backbone could have been used as well, though pSB3T5 gave us more flexibility with the construct as additional regulatory elements were not required such as the case with pSB2K3. CharacterizationStrategyResults |
