Boston UniversityPlasmidChoice

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(Plasmid Choice)
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== Plasmid Choice==
== Plasmid Choice==
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Our plan is to replicate our engineered plasmid in E. coli and then transfer the plasmid to S. oneidensis through conjugation.  In order to allow for conjugation, an oriT site must be present.  We have chosen plasmid PJQ200 because it has this site.  The normal form of PJQ200 contains a sacB site, which in the presence of sucrose is lethal in non-enterobacteria (and S. oneidensis is not an enterobacterium).  In order to not harm S. oneidensis after the conjugation, the plasmid will be engineered beforehand so the sacB site is not present.
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Our plan is to replicate our engineered plasmid in E. coli and then transfer the plasmid to S. oneidensis through conjugation.  In order to allow for conjugation, an oriT site must be present, and we have chosen to use the plasmid PJQ200 because it has this site.  The normal form of PJQ200 contains a sacB site, which in the presence of sucrose is lethal in non-enterobacteria (and S. oneidensis is not an enterobacterium).  In order to not harm S. oneidensis after the conjugation, the plasmid will be engineered beforehand so the sacB site is not present.

Revision as of 15:12, 8 June 2007

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Plasmid Choice

Our plan is to replicate our engineered plasmid in E. coli and then transfer the plasmid to S. oneidensis through conjugation. In order to allow for conjugation, an oriT site must be present, and we have chosen to use the plasmid PJQ200 because it has this site. The normal form of PJQ200 contains a sacB site, which in the presence of sucrose is lethal in non-enterobacteria (and S. oneidensis is not an enterobacterium). In order to not harm S. oneidensis after the conjugation, the plasmid will be engineered beforehand so the sacB site is not present.