Team:Cornell/testing/project/wetlab/5
From 2012.igem.org
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- | <h3> Chromosomal Integration </h3> | + | </div> |
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+ | <h3>Chromosomal Integration </h3> | ||
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. | 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. | 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. | ||
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+ | <h3> Proteolysis Tag </h3> | ||
+ | Proteins can be tagged for degradation by proteases with a proteolysis tag. Thus, by fusing a proteolysis tag to mtrB, mtrB will be degraded at a constant rate, allowing us to lower the steady state presence of mtrB. As well as eliminating noisy current generated via leaky expression, this would make it less likely that upregulating mtrB will reach a saturated current level. | ||
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Revision as of 16:56, 2 October 2012