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Team:Minnesota/Parts Characterization
From 2009.igem.org
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Parts Characterization
We chose five promoters that were included in the 2009 iGEM Kit to characterize. Using part BBa_F2620, which was characterized by a group from MIT in 2004, as a template, we characterized the following parts:
| Part | Description | Regulators | People |
|---|---|---|---|
| I14032 | Constitutive promoter classified as repressible | IPTG | Princeton 2004 |
| J13002 | Two TetR binding sites and RBS | aTc | UT Austin 2005 |
| I14015 | LasR, 3OC12HSL aTc regulated promoter | LasR, 3C12HSL, aTc | Princeton 2004 |
| K091101 | TTL AND gate | IPTG, aTc | Davidson Missouri-Western 2008 |
| R0011 | Inverting regulatory region controlled by LacI; for comparison since already characterized | IPTG | Registry |
We chose these parts because they had the same regulators as the promoters we examined in our project. Part K091101 was particularly interesting because it was one of the constructs of the Tet and Lac operators that we examined for our research. Since Tet and Lac are commonly studied operators in synthetic biology, we wanted parts that involved them to be well-characterized to ensure the viability of future research.
We decided to characterize these promoters by attaching them to part K081012, which consists of a strong RBS and GFP. This 'PoPS generator' takes PoPS (Polymerase Per Second) as an input and gives GFP as an output, allowing us to indirectly measure PoPS and characterize our parts.
The 2009 iGEM Judging Criteria gives MIT's characterization of part BBa_F2620 as an exemplar of parts characterization. We gratefully acknowledge their pioneering work with parts characterization and hope that our work continues to maintain the high standard for characterization of parts.
