Team:Hong Kong HKUST/pneumosensor/future work
From 2014.igem.org
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<div class='content_1'><h3> Lysis Module </h3> | <div class='content_1'><h3> Lysis Module </h3> | ||
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Our team designed but was unable to test this module due to the limitation of not being able to work directly with <i>Streptococcus | Our team designed but was unable to test this module due to the limitation of not being able to work directly with <i>Streptococcus | ||
Pneumoniae</i> (Biosafety level 2) in our lab. This module proposes to kill <i>Streptococcus Pneumoniae</i> upon detection when coupled | Pneumoniae</i> (Biosafety level 2) in our lab. This module proposes to kill <i>Streptococcus Pneumoniae</i> upon detection when coupled | ||
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<p> The enzymes are tagged with osmY (Washington 2012) via a linker to be exported out of <i>Escherichia coli</i>. Both enzymes have | <p> The enzymes are tagged with osmY (Washington 2012) via a linker to be exported out of <i>Escherichia coli</i>. Both enzymes have | ||
very different N-terminal catalytic sites and share a similar C-terminal cell wall attachment site, which binds to choline in both cases. | very different N-terminal catalytic sites and share a similar C-terminal cell wall attachment site, which binds to choline in both cases. | ||
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membrane and ultimate lysis of <i>S. pneumoniae</i>. | membrane and ultimate lysis of <i>S. pneumoniae</i>. | ||
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- | + | <p> 1. Put inducible promoter upstream of RBS (BBa_B0034), comX gene, and terminator (BBa_B0015). An example of inducible promoter is BBa_I0500. Hence, by putting an inducible promoter, we can tune the level of comX protein expression and characterize combox promoters (PcelA and Phelicase) on different level of comX concentration. | |
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- | 1. Put inducible promoter upstream of RBS (BBa_B0034), comX gene, and terminator (BBa_B0015). An example of inducible promoter is BBa_I0500. Hence, by putting an inducible promoter, we can tune the level of comX protein expression and characterize combox promoters (PcelA and Phelicase) on different level of comX concentration. | + | |
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+ | <u>References</u> | ||
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+ | J.M. Loeffler et al "Rapid Killing of <i>Streptococcus pneumoniae</i> with a Bacteriophage Cell Wall Hydrolase" 2001 | ||
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+ | J.M. Loeffler et al "Phage Lytic Enzyme Cpl-1 as a Novel Antimicrobial for Pneumococcal Bacteremia" 2003 | ||
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+ | J.M. Loeffler et al "Synergistic Lethal Effect of a Combination of Phage Lytic Enzymes with Different Activities on Penicillin-Sensitive | ||
+ | and -Resistant <i>Streptococcus pneumoniae</i> Strains" 2003 | ||
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Revision as of 10:09, 4 October 2014
Pneumosensor Future Work
Lysis Module
Our team designed but was unable to test this module due to the limitation of not being able to work directly with Streptococcus Pneumoniae (Biosafety level 2) in our lab. This module proposes to kill Streptococcus Pneumoniae upon detection when coupled with the detection and regulation modules by releasing specific bacteriophage lytic enzymes, Cpl-1 and Pal.
The enzymes are tagged with osmY (Washington 2012) via a linker to be exported out of Escherichia coli. Both enzymes have very different N-terminal catalytic sites and share a similar C-terminal cell wall attachment site, which binds to choline in both cases. Cleavage with either of these enzymes results in a weakening in the cell wall, which leads to the externalization of the cytoplasmic membrane and ultimate lysis of S. pneumoniae. |
ComX, PcelA, Phelicase Future Work
1. Put inducible promoter upstream of RBS (BBa_B0034), comX gene, and terminator (BBa_B0015). An example of inducible promoter is BBa_I0500. Hence, by putting an inducible promoter, we can tune the level of comX protein expression and characterize combox promoters (PcelA and Phelicase) on different level of comX concentration.
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References
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