Team:MIT phage design

From 2010.igem.org

(Difference between revisions)
Line 58: Line 58:
<div id="unique" style="padding:0px; font-size: 14px; border: 1px solid black; margin:0px; background-color:transparent;">
<div id="unique" style="padding:0px; font-size: 14px; border: 1px solid black; margin:0px; background-color:transparent;">
-
<table width=650px style="background-color: white; margin-top:5px; padding: 10px;"><tr><td><div class="bodybaby">hairy cells and polymerizing phage</div></td>
+
<table width=650px style="background-color: white; margin-top:5px; padding: 10px;"><tr><td><div class="bodybaby">hairy cells and polymerizing phage - design</div></td>
<tr><td>
<tr><td>
<br>
<br>
-
<b>DESIGN</b>
+
<b>P8-FUSION DESIGN</b>
<br>
<br>
<img src="https://static.igem.org/mediawiki/2010/f/fa/Phage_design.png" width=630px>
<img src="https://static.igem.org/mediawiki/2010/f/fa/Phage_design.png" width=630px>

Revision as of 18:20, 24 October 2010

hairy cells and polymerizing phage - design

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.)