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Team:Berkeley Wetlab/Passenger: Leucine Zippers
From 2009.igem.org
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EILD, leucine zipper (14)<br> | EILD, leucine zipper (14)<br> | ||
KILR, leucine zipper (14)<br> | KILR, leucine zipper (14)<br> | ||
| + | EILD + KILR (14) (EILD can heterodimerize with KILR)<br> | ||
1363 negative control (1) (Periplasm targeted strepavidin binding protein)<br> | 1363 negative control (1) (Periplasm targeted strepavidin binding protein)<br> | ||
CPG L2 positive control(1) (Leucine zipper previously shown to cause cell-cell adhesion<br> | CPG L2 positive control(1) (Leucine zipper previously shown to cause cell-cell adhesion<br> | ||
Revision as of 04:39, 21 October 2009
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Contents |
Leucine Zippers
One potential use of cell surface display is to adhere cells together selectively such that there is structure. However, such a feat is impossible to do in one step. In order to work up to such a goal, the first step of such an endeavor is create a basic system of adhere 2 cells types together. We propse using dimeric leucine zippers to adhere cells to one another. Leucine zippers are short peptides which form alpha helices that compliment each other. Using a GNC4 leucine zipper heterodimers KILR and EILD, we hoped to adhere cells to one another.
Functional Assay: Cell-Cell Adhesion
This assay tests for the functionality of displayed leucine zippers via the leucine zipper's ability to dimerized and, as a result, pull the cells out of solution into a clump.
Constructs:
EILD, leucine zipper (14)
KILR, leucine zipper (14)
EILD + KILR (14) (EILD can heterodimerize with KILR)
1363 negative control (1) (Periplasm targeted strepavidin binding protein)
CPG L2 positive control(1) (Leucine zipper previously shown to cause cell-cell adhesion
All experiments are done in triplicate
cell growth and induction
