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Team:Tec-Monterrey/projectmodeling/construct2
From 2011.igem.org
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| - | + | Our most promising construct was our first “Extracellular Expression Device”, mediated by an Auto-transporter membrane protein complex. For this device we characterized two new parts, and was made up of five parts. We used an arabinose induced promoter Pbad (BBa_K206000) and its repressor enzyme AraC (BBa_I13458), followed next by one of our first part BBa_K633002, made out of Ribosome binding site (BBa_b0034) and our signal peptide phoA and enzyme cellulase. Our second part BBa_K6330014, the extracellular expressing complex made out of a linker followed by our auto-transporter membrane protein estA. | |
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| + | Our Signal Peptide “phoA” was selected based upon the former usage and correct operation in the expression of extracellular lipolytic enzymes in e. coli’s periplasm. | ||
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| + | Our construction begins with the signal peptide from an e. coli phosphatase, which is the first recognizable segment from our sequence and directs the polypeptide chain to the periplasm, followed by our membrane protein estA, formerly used to transport and maintain attached lipase outside gram negative bacteria. Since this construct has its C terminal exported outside the cell we needed to begin our 5’ sequence with our exterior domain, our modified cellulase, which has a modified hystidil residue on its active site giving it a augmenting its activity in a 200%. | ||
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Revision as of 03:57, 29 September 2011
Our most promising construct was our first “Extracellular Expression Device”, mediated by an Auto-transporter membrane protein complex. For this device we characterized two new parts, and was made up of five parts. We used an arabinose induced promoter Pbad (BBa_K206000) and its repressor enzyme AraC (BBa_I13458), followed next by one of our first part BBa_K633002, made out of Ribosome binding site (BBa_b0034) and our signal peptide phoA and enzyme cellulase. Our second part BBa_K6330014, the extracellular expressing complex made out of a linker followed by our auto-transporter membrane protein estA.
Our Signal Peptide “phoA” was selected based upon the former usage and correct operation in the expression of extracellular lipolytic enzymes in e. coli’s periplasm.
Our construction begins with the signal peptide from an e. coli phosphatase, which is the first recognizable segment from our sequence and directs the polypeptide chain to the periplasm, followed by our membrane protein estA, formerly used to transport and maintain attached lipase outside gram negative bacteria. Since this construct has its C terminal exported outside the cell we needed to begin our 5’ sequence with our exterior domain, our modified cellulase, which has a modified hystidil residue on its active site giving it a augmenting its activity in a 200%.
