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Team:MIT phage design
From 2010.igem.org
iGEM 2010
hairy cells and polymerizing phage - design |
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P8-FUSION DESIGN 
The above shows the RBS + p8-fusion genetic design. We chose p8 because of the large number of copies, increasing the potential for incorporation, and hopefully allowing for large-scale polymerization instead of small-scale clumping which might happen with p9 (due to the small number of p9 produced). This genetic fusion is designed to allow the expression of a leucine zipper (coil in the diagram) on the p8 phage coat protein. There are six total p8 fusions for the six different zippers. To assist in proper cellular trafficking to the membrane, a leader sequence was introduced. Additionally, HA and Myc tags were introduced (one for each half of a zipper pair), which were used in western blotting experiments to test for the expression of the p8-fusion genes. A special version of p8 has been used, called opti-p8, which displays proteins better (see Weiss et al. Mutational analysis of the major coat protein of M13 identifies residues that control protein display. 2000.) PHAGE-RELEVANT CIRCUITRY DESIGN 
P8-FUSION DESIGN IN DETAIL Below is a PDF document detailing all aspects of the fusion designs, including some design notes and references. |
