Team:Cornell/testing/project/wetlab/2

From 2012.igem.org

Revision as of 07:09, 3 October 2012 by D.Webster (Talk | contribs)

Chassis

Intro to electroactivity/Shewanella/Mtr pathway

Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.

CymA

Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.

MtrA, MtrB

Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.

MtrC and OmcA

Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.

Role of mtrB

A small plasmid with few expressed genes may not affect the current output of S. oneidensis to a significant degree, but a large plasmid with many expressed genes (such as our secondary naphthalene degradation plasmid) significantly impairs the growth and metabolism of S. oneidensis. Integrating the naphthalene degradation operon into the chromosome of S. oneidensis may help partially alleviate the energy cost of replicating several copies of a huge plasmid. In addition to alleviating the stress caused by expressing a giant operon, integrating our genetic parts into the chromosome eliminates the need to design a selective pressure for S. oneidensis to maintain extrachromosomal DNA.

References

Proteins can be tagged for degradation by proteases with a proteolysis tag. By fusing such a tag to MtrB, we can tell the cell to degrade the protein at a higher rate, allowing us to decrease the steady state concentration of MtrB at all levels of analyte. If we are able to tune the degradation of MtrB such that its concentration at uninduced levels is not sufficient to complex with available MtrA and MtrC, the basal current production that our engineered strains produce would be decreased. Consequently, the dynamic range of our biosensor would be increased, since higher levels of analyte would be needed to generate the promoter activity requisite to produce MtrB in sufficient quantity to fully localize all MtrA and MtrC—i.e., current production would saturate at higher levels of analyte.