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Team:Hong Kong HKUST/pneumosensor/future work
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| - | <p>P<sub>comCDE</sub> is a promoter induced by phosphorylated ComE, which can be replaced by a phosphorylmimetic mutant (ComE<sup> | + | <p>P<sub>comCDE</sub> is a promoter induced by phosphorylated ComE, which can be replaced by a phosphorylmimetic mutant (ComE<sup>D58E</sup>) in experiment. We could not |
manage to verify the sequence of our ligated product, P<sub>comCDE</sub>- BBa_E0240. Therefore, in the future, we hope to continue the verification process, and complete our | manage to verify the sequence of our ligated product, P<sub>comCDE</sub>- BBa_E0240. Therefore, in the future, we hope to continue the verification process, and complete our | ||
construct.</p> | construct.</p> | ||
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and -Resistant <i>Streptococcus pneumoniae</i> Strains" 2003 | and -Resistant <i>Streptococcus pneumoniae</i> Strains" 2003 | ||
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Latest revision as of 15:44, 14 November 2014
Pneumosensor Future Work
Lysis Module
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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 Streptococcus Pneumoniae (Biosafety level 2) in our lab. This module proposes to kill Streptococcus Pneumoniae upon detection when coupled with the detection and modules by releasing specific bacteriophage lytic enzymes, Cpl-1 and Pal.
The enzymes are tagged with osmY (BBa_K892008, 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. |
PcomCDE
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PcomCDE is a promoter induced by phosphorylated ComE, which can be replaced by a phosphorylmimetic mutant (ComED58E) in experiment. We could not manage to verify the sequence of our ligated product, PcomCDE- BBa_E0240. Therefore, in the future, we hope to continue the verification process, and complete our construct. |
σx, PcelA, PcomFA
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PcelA and PcomFA promoters are promoter that is regulated by σx. We have proven that in the presence of σx, PcelA and PcomFA could express GFP. However, we did not manage to characterize the GFP expression of PcelA and PcomFA in different concentration of σx. So, a possible future work is to put inducible promoter upstream of RBS (BBa_B0034), σx 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 σx expression and characterize Com-Box promoters (PcelA and PcomFA) on different level of σx concentration.
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