Difference between revisions of "Team:Pitt/3-hybrid/Project"
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<div class="color"><h4>Project Background</h4>This project aims to develop the full versatility that paper-based sensors can have. This system uses the idea of a three-hybrid system, where one part binds the promoter, another part is a subunit of <i>E. coli</i> RNA Polymerase, and the analyte of choice provides the bridge that recruits the RNAP to the DNA. For this system, we chose two analytes: VEGF-A, which is a small dimeric protein involved in many cancers, and anti-MUC1 antibodies, which are present in many cancer patients. Both of these sensors use the same DNA binding domains and RNA Polymerase domain, inspired by a bacterial two-hybrid system developed in 2000. (<a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC16554/">Joung 2000</a>) In fact, the only difference between the sensors are the proteins fused to these domains. In the VEGF-A sensor, we used a single chain variable fragment antibody, which is extremely specific to its target. (<a href="http://www.sciencedirect.com/science/article/pii/S0022283699931923">Chen 1999</a>) In the anti-MUC1 antibody sensor, the bait is a portion of the MUC1 protein, as shown in the image below.<br/><img style="width:85%" src="https://static.igem.org/mediawiki/2015/5/52/Pitt6.png"/></div> | <div class="color"><h4>Project Background</h4>This project aims to develop the full versatility that paper-based sensors can have. This system uses the idea of a three-hybrid system, where one part binds the promoter, another part is a subunit of <i>E. coli</i> RNA Polymerase, and the analyte of choice provides the bridge that recruits the RNAP to the DNA. For this system, we chose two analytes: VEGF-A, which is a small dimeric protein involved in many cancers, and anti-MUC1 antibodies, which are present in many cancer patients. Both of these sensors use the same DNA binding domains and RNA Polymerase domain, inspired by a bacterial two-hybrid system developed in 2000. (<a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC16554/">Joung 2000</a>) In fact, the only difference between the sensors are the proteins fused to these domains. In the VEGF-A sensor, we used a single chain variable fragment antibody, which is extremely specific to its target. (<a href="http://www.sciencedirect.com/science/article/pii/S0022283699931923">Chen 1999</a>) In the anti-MUC1 antibody sensor, the bait is a portion of the MUC1 protein, as shown in the image below.<br/><img style="width:85%" src="https://static.igem.org/mediawiki/2015/5/52/Pitt6.png"/></div> | ||
Latest revision as of 01:28, 19 September 2015
Three-Hybrid Versatile Sensor
Project Background
This project aims to develop the full versatility that paper-based sensors can have. This system uses the idea of a three-hybrid system, where one part binds the promoter, another part is a subunit of E. coli RNA Polymerase, and the analyte of choice provides the bridge that recruits the RNAP to the DNA. For this system, we chose two analytes: VEGF-A, which is a small dimeric protein involved in many cancers, and anti-MUC1 antibodies, which are present in many cancer patients. Both of these sensors use the same DNA binding domains and RNA Polymerase domain, inspired by a bacterial two-hybrid system developed in 2000. (Joung 2000) In fact, the only difference between the sensors are the proteins fused to these domains. In the VEGF-A sensor, we used a single chain variable fragment antibody, which is extremely specific to its target. (Chen 1999) In the anti-MUC1 antibody sensor, the bait is a portion of the MUC1 protein, as shown in the image below.