Team:MIT phage construction

From 2010.igem.org

Phage
hairy cells and polymerizing phage - construction

PARTS

BBa_K415100
  • Components: RBS : M13 PVIII
  • Explanation: This is a basic translational unit: a ribosome-binding site and the p8 gene. You can attach any promoter in front of it for expression.



BBa_K415108
  • Components: RBS : M13 PIII
  • Explanation: This is a basic translational unit: a ribosome-binding site and the p3 gene. You can attach any promoter in front of it for expression.



BBa_K415138
  • Components: pLac : RBS : M13 PIII
  • Explanation: This is a basic expression unit: a promoter in front of a ribosome-binding site and the p3 gene. We used this to generate constitutive expression of p3 in cells infected with hyperphage in an attempt to rescue the wild-type, non-hairy phenotype. See the results page for an AFM image of this working.



BBa_K415147 - K415152
  • Components: PluxR/cI : RBS : mCherry : Term : PtetR : RBS : LuxR:Term : Plux/cI : RBS : M13 PVIII-Zipper
  • Explanation: These parts are K415010 + RBS + P8-fusion protein. As is, this part produces mCherry and the p8-fusion gene in the presence of c6-AHL (and there's also non-trivial leaky expression). When combined with the Hyperphage plasmid, the p8-fusion protein should become incorporated into the phage coat along with normal p8; zippers should then be exposed on the polyphage hairs outside of the cell, allowing for potential cross-linking among cells. With the Collins toggle, UV exposure can be used to control the expression of the p8-fusion protein. Note: below is K415147 only; the rest are similar but with a different translational unit on the end for each linker (Fos, Jun, ACID, BASE, GR1, GR2).

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