Team:MIT phage construction

From 2010.igem.org

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<br><br><b>BBa_K415147 - K415152</b>
<br><br><b>BBa_K415147 - K415152</b>
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Components: PluxR/cI : RBS : mCherry : Term : PtetR : RBS : LuxR:Term : Plux/cI : RBS : M13 PVIII-Zipper/li>
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Components: PluxR/cI : RBS : mCherry : Term : PtetR : RBS : LuxR:Term : Plux/cI : RBS : M13 PVIII-Zipper</li>
<li>Explanation: These parts are K415010 + RBS + P8-fusion protein.  As is, this part produces mCherry and the p8-fusion protein 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, along for the zippers to 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.
<li>Explanation: These parts are K415010 + RBS + P8-fusion protein.  As is, this part produces mCherry and the p8-fusion protein 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, along for the zippers to 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.
</li></ul>
</li></ul>

Revision as of 23:41, 24 October 2010

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 protein 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, along for the zippers to 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.

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