IGEM:MIT/2005/End of Week Meeting: Friday, July 8

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ToxR

  • N-term has restriction site
    • Get rid of it, where it might not work, but it might.
  • May autodimerize; multiple places that mediate this.
  • See Jappelli & Brenner
    • Abstract: In order to assess the potentiality of Vibrio cholerae ToxR protein and of bacteriophage small lambda, Greek repressor as indicators of the dimerization of periplasmic proteins in Escherichia coli, we have constructed a series of plasmids encoding transmembrane fusion proteins. The amino-terminal part, containing the DNA binding domain of either ToxR or small lambda, Greek repressor, is located in the cytoplasm and acts as reporter for dimerization. As models of periplasmic proteins we have used alkaline phosphatase (a dimer) and small beta, Greek-lactamase (a monomer). Both the expression level and the distance between the transmembrane segment and the periplasmic protein substantially affect the activity of the reporter domains.
  • Mary Berlyn - runs E.coli genomic stock center, Yale
  • pSB1A2

Goals

  • Spec out all parts in registry.
  • Design some primers! Order them!