Difference between revisions of "Team:William and Mary/RealMainPage"

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         Although Cameron and Collins already demonstrated the functionality of protein degradation tags (pdt), and the Mesoplasma florum Lon (mf-Lon) protease in E. coli, we noted that they did their work exclusively using genomically integrated constructs. Since the majority of iGEM teams work exclusively or close to exclusively on plasmid constructs, we first wanted to confirm and characterize the parts using iGEM backbones. To do this we assembled constitutive and ATC
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inducible constructs carrying the red fluorescent protein mScarlet-I, tagged with each of our six different pdts, or left untagged as a control. Further, to ensure that our project will work with a variety of different proteins, we made identical constructs encoding for superfolder GFP (sfGFP), and preformed preliminary characterization.
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Revision as of 21:34, 31 October 2017




This part is designed to easily facilitate appending the protein degradation tag (pdt)#3 tag to the end of an arbitrary protein using Gibson assembly, without requiring multiple cloning steps. UNS pdt#3 DT contains a tail that can be degrade Mesoplasma florum’s Lon protease, which is orthogonal to E. Coli’s own degradation machinery. As this part contains both a double stop codon and the B0015 double terminator, it can be added before the stop codons of an arbitrary protein, preventing a multistep assembly to incorporate double stop codons and a double terminator. Of this pdt tag series, this part has the highest degradation rate.


Although Cameron and Collins already demonstrated the functionality of protein degradation tags (pdt), and the Mesoplasma florum Lon (mf-Lon) protease in E. coli, we noted that they did their work exclusively using genomically integrated constructs. Since the majority of iGEM teams work exclusively or close to exclusively on plasmid constructs, we first wanted to confirm and characterize the parts using iGEM backbones. To do this we assembled constitutive and ATC inducible constructs carrying the red fluorescent protein mScarlet-I, tagged with each of our six different pdts, or left untagged as a control. Further, to ensure that our project will work with a variety of different proteins, we made identical constructs encoding for superfolder GFP (sfGFP), and preformed preliminary characterization.




Database
Upcoming


Database
Upcoming




what happens
Standard example of inline elements This is some text.


This is an example of a right alignment. This is some text.


This is example of a left alignment This is some text.










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This part is designed to easily facilitate appending the protein degradation tag (pdt)#3 tag to the end of an arbitrary protein using Gibson assembly, without requiring multiple cloning steps. UNS pdt#3 DT contains a tail that can be degrade Mesoplasma florum’s Lon protease, which is orthogonal to E. Coli’s own degradation machinery. As this part contains both a double stop codon and the B0015 double terminator, it can be added before the stop codons of an arbitrary protein, preventing a multistep assembly to incorporate double stop codons and a double terminator. Of this pdt tag series, this part has the highest degradation rate. This part is designed to easily facilitate appending the protein degradation tag (pdt)#3 tag to the end of an arbitrary protein using Gibson assembly, without requiring multiple cloning steps. UNS pdt#3 DT contains a tail that can be degrade Mesoplasma florum’s Lon protease, which is orthogonal to E. Coli’s own degradation machinery. As this part contains both a double stop codon and the B0015 double terminator, it can be added before the stop codons of an arbitrary protein, preventing a multistep assembly to incorporate double stop codons and a double terminator. Of this pdt tag series, this part has the highest degradation rate. This part is designed to easily facilitate appending the protein degradation tag (pdt)#3 tag to the end of an arbitrary protein using Gibson assembly, without requiring multiple cloning steps. UNS pdt#3 DT contains a tail that can be degrade Mesoplasma florum’s Lon protease, which is orthogonal to E. Coli’s own degradation machinery. As this part contains both a double stop codon and the B0015 double terminator, it can be added before the stop codons of an arbitrary protein, preventing a multistep assembly to incorporate double stop codons and a double terminator. Of this pdt tag series, this part has the highest degradation rate.




This part is designed to easily facilitate appending the protein degradation tag (pdt)#3 tag to the end of an arbitrary protein using Gibson assembly, without requiring multiple cloning steps. UNS pdt#3 DT contains a tail that can be degrade Mesoplasma florum’s Lon protease, which is orthogonal to E. Coli’s own degradation machinery. As this part contains both a double stop codon and the B0015 double terminator, it can be added before the stop codons of an arbitrary protein, preventing a multistep assembly to incorporate double stop codons and a double terminator. Of this pdt tag series, this part has the highest degradation rate. This part is designed to easily facilitate appending the protein degradation tag (pdt)#3 tag to the end of an arbitrary protein using Gibson assembly, without requiring multiple cloning steps. UNS pdt#3 DT contains a tail that can be degrade Mesoplasma florum’s Lon protease, which is orthogonal to E. Coli’s own degradation machinery. As this part contains both a double stop codon and the B0015 double terminator, it can be added before the stop codons of an arbitrary protein, preventing a multistep assembly to incorporate double stop codons and a double terminator. Of this pdt tag series, this part has the highest degradation rate. This part is designed to easily facilitate appending the protein degradation tag (pdt)#3 tag to the end of an arbitrary protein using Gibson assembly, without requiring multiple cloning steps. UNS pdt#3 DT contains a tail that can be degrade Mesoplasma florum’s Lon protease, which is orthogonal to E. Coli’s own degradation machinery. As this part contains both a double stop codon and the B0015 double terminator, it can be added before the stop codons of an arbitrary protein, preventing a multistep assembly to incorporate double stop codons and a double terminator. Of this pdt tag series, this part has the highest degradation rate.




This part is designed to easily facilitate appending the protein degradation tag (pdt)#3 tag to the end of an arbitrary protein using Gibson assembly, without requiring multiple cloning steps. UNS pdt#3 DT contains a tail that can be degrade Mesoplasma florum’s Lon protease, which is orthogonal to E. Coli’s own degradation machinery. As this part contains both a double stop codon and the B0015 double terminator, it can be added before the stop codons of an arbitrary protein, preventing a multistep assembly to incorporate double stop codons and a double terminator. Of this pdt tag series, this part has the highest degradation rate. This part is designed to easily facilitate appending the protein degradation tag (pdt)#3 tag to the end of an arbitrary protein using Gibson assembly, without requiring multiple cloning steps. UNS pdt#3 DT contains a tail that can be degrade Mesoplasma florum’s Lon protease, which is orthogonal to E. Coli’s own degradation machinery. As this part contains both a double stop codon and the B0015 double terminator, it can be added before the stop codons of an arbitrary protein, preventing a multistep assembly to incorporate double stop codons and a double terminator. Of this pdt tag series, this part has the highest degradation rate. This part is designed to easily facilitate appending the protein degradation tag (pdt)#3 tag to the end of an arbitrary protein using Gibson assembly, without requiring multiple cloning steps. UNS pdt#3 DT contains a tail that can be degrade Mesoplasma florum’s Lon protease, which is orthogonal to E. Coli’s own degradation machinery. As this part contains both a double stop codon and the B0015 double terminator, it can be added before the stop codons of an arbitrary protein, preventing a multistep assembly to incorporate double stop codons and a double terminator. Of this pdt tag series, this part has the highest degradation rate.