Summary and Outline of 2011 Yale iGEM (PPT)
In the past month, we have:
1) Successfully expressed four different antifreeze protein constructs in BL21 and Origami strain E. coli cells (verified by observing green pellets, fluorimetry, SDS/PAGE Gel electrophoresis, and Western blotting)
2) Demonstrated enhanced cold survivability at -20° C in cells expressing antifreeze protein (microbial plating assay)
3) Verified sequences of all antifreeze protein constructs
4) Cloned three antifreeze protein constructs into the standard iGEM BioBrick vector
We have received the synthetic novel Siberian beetle antifreeze protein gene that was sponsored by IDT and have begun cloning and subsequent experiments on this construct. We are also developing an ice recrystallization inhibition assay and finishing the cloning of several more BioBricks. Soon, we hope to begin optimizing expression of the antifreeze proteins using directed evolution technologies.
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