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Project Modules

[Under construction]

In order to successfully create a cold resistant strain of E. coli, we intend to express and combine the chaperones and AFPs listed above.

Since all action in a cell grinds to a halt at low temperatures due to reduced protein function it is essential for us to use a promoter that can effectively operate at low temperatures. Isoude. Even though some of our targets are documented to convey increased cold growth we will still have to confirm this. Moreover, our other genes of interest may have an influence on cold growth as well and, as such, will have to be characterized in the same way. Similarly, the effect of our targets conveying resistance to repeated freezing and thawing will have to be validated as well. For this experiment we will once again study all of our targets to see if any undocumented effects emerge.

The Project was divvied up into several project modules. Note that the first module has to be executed independently, allowing for the second and third to be executed in tandem, modules 4 and five can be started as soon as the first three modules are done. The final item can be performed based on the information gained from the fourth point.

Characterizing and modeling promoters

1. • assemble a number of promoters with a strong Ribosome binding site and either a GFP or B-gal reporter
   • Isoude

Characterizing growth rate of individual Growth rate enhancing, antifreeze & freeze resistance proteins

1. • Use a promoter that has been shown to effectively (over?)express GFP or B-gal
   • culture induced cells at 37°C and below, measuring OD600 at set intervals

Characterizing freeze resistance proteins on cell viability

1. • Bas

Combining BioBricks based on their characterization in order to maximize growth rate

1. • Assemble combinations of two working BioBricks into a single plasmid
   • culture induced cells at 37°C and below, measuring OD600 at set intervals
2. Combining Growth rate enhancing with antifreeze for subzero characterization
   • Assemble combinations of two working BioBricks into a single plasmid; a growth-rate enhancer that has been shown to allow growth at 4°C with an antifreeze protein
3. Make and test a cold resistance backbone
   • Bas