Team:Berkeley Wetlab/Passenger: Leucine Zippers

Contents 1 Leucine Zippers 2 Functional Assay: Cell-Cell Adhesion 2.1 cell growth and induction 2.2 vortex and take ODs 3 Results 3.1 Data 3.2 Conclusions

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

1. Grow a cells of each typein a liquid culture overnight to saturation.
2. Make a 1:1000 dilution the following day of the saturated cultures and innoculate with arabinose (100 μg/mL final concentration) and incubate for 24-48 hours. For the the EILD + KILR sample add both EILD and KILR to grow both cell types in the same tube.

vortex and take ODs

1.

Results

Data

Conclusions

• Recent changes
• What links here
• Related changes
• Special pages
• My preferences

• Printable version
• Permanent link
• Privacy policy
• Disclaimers