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Team:WPI-Worcester/Our-Construct
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| - | <h4>Our Construct</h4><p><center><img src="https://static.igem.org/mediawiki/2014/thumb/e/e0/WPI_OurConstruct.png/800px-WPI_OurConstruct.png"/></center></p><p>The construct developed in this project is a combination of the N-terminal domain of BclA and the CAEV p28 antigen. The sequence encoding the N-terminal domain of BclA is ligated to the sequence for the CAEV p28 antigen. This plasmid is then combined with a ribosome binding site, a constitutive promoter, and a double terminator in order to make a complete construct. The resulting plasmid allows the <i>E. coli</i> it is transformed into to express the CAEV p28 antigen connected to the N-terminal domain of BclA, which is anchored in the outer surface of the plasma membrane. This plasmid is transformed into competent <i>E. coli</i>, which then express CAEV on their membrane surface, thanks to the anchoring of BclA’s N-terminal domain. | + | <h4>Our Construct</h4><p><center><img src="https://static.igem.org/mediawiki/2014/thumb/e/e0/WPI_OurConstruct.png/800px-WPI_OurConstruct.png"/></center></p><p>The construct developed in this project is a combination of the N-terminal domain of BclA and the CAEV p28 antigen. The sequence encoding the N-terminal domain of BclA is ligated to the sequence for the CAEV p28 antigen. Both sequences were ordered as codon-optimized oligos, so that <i>B. anthracis</i> and CAEV would not have to be worked with directly. This plasmid is then combined with a ribosome binding site, a constitutive promoter, and a double terminator in order to make a complete construct. The resulting plasmid allows the <i>E. coli</i> it is transformed into to express the CAEV p28 antigen connected to the N-terminal domain of BclA, which is anchored in the outer surface of the plasma membrane. This plasmid is transformed into competent <i>E. coli</i>, which then express CAEV on their membrane surface, thanks to the anchoring of BclA’s N-terminal domain. |
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Revision as of 04:16, 17 October 2014
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Team:WPI-Worcester
From 2014.igem.org
Our Construct
The construct developed in this project is a combination of the N-terminal domain of BclA and the CAEV p28 antigen. The sequence encoding the N-terminal domain of BclA is ligated to the sequence for the CAEV p28 antigen. Both sequences were ordered as codon-optimized oligos, so that B. anthracis and CAEV would not have to be worked with directly. This plasmid is then combined with a ribosome binding site, a constitutive promoter, and a double terminator in order to make a complete construct. The resulting plasmid allows the E. coli it is transformed into to express the CAEV p28 antigen connected to the N-terminal domain of BclA, which is anchored in the outer surface of the plasma membrane. This plasmid is transformed into competent E. coli, which then express CAEV on their membrane surface, thanks to the anchoring of BclA’s N-terminal domain.
