Team:Virginia

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University of Virginia duPont

Transcription attenuation for metabolic control by engineering intrinsic terminators

A main challenge in constructing synthetic biological systems is the inability to precisely regulate gene expression using artificial means. Tightly-regulated control of any given set of related transcriptional, translational and posttranslational events will likely require a combination of powerful strategies. Therefore, the 2008 Virginia iGEM team is developing a library of transcriptional terminators intentionally redesigned to be functionally inefficient. Well-characterized, standardized terminators of various efficiencies should allow finely-tuned transcription attenuation and represents yet another step toward global biological control. This work complements other gene expression control methods that focus on initiation of transcription. The desired result is quantitative control of transcript levels, which is often necessary to balance flux through a synthetic metabolic pathway. To demonstrate its potential for real-world application, the team is planning to employ this approach to control the expression of a heterologous pathway in E. coli for the biosynthesis of polyhydroxybutyrate (PHB), a biodegradable polyester plastic.

Genetic Attenuators

Getting in control of transcription

Terminators are never 100% efficient, meaning that not all polymerases will disengage from the strand they are operating on. This phenomenon can be exploited to create Genetic Attentuators. Inserting a Genetic Attenuator between genes will create transcripts containing different sets of genes. Varying the number of copies of a gene present in the transcript will influence downstream translation of that gene.

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Placeholder Sites

Cloning sites made easy!

Assembling gene constructs is a tedious procedure that consumes valuable time. When assembling several BioBricks into a vector it would be valuable to be able to insert a cloning site which could later be used to insert a BioBrick into a certain part of the vector. Placeholder Sites accomplish this task by providing restriction sites compatible with the BioBrick standard inside of a standard part.

  1. EX - ApoI (NotI) AvrII - SP
  2. EX - ApoI (NotI) NheI - SP
  3. EX - ApoI (NotI) NsiI - SP
  4. EX - ApoI (NotI) SbfI - SP
  5. EX - MfeI (NotI) AvrII - SP
  6. EX - MfeI (NotI) NheI - SP
  7. EX - MfeI (NotI) Nsil - SP
  8. EX - MfeI (NotI) SbfI - SP
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BioPlastic

Growing a renewable resource

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Additions to the Registry

Giving back

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