Team:Yale/Project Validate
From 2013.igem.org
Project Overview | Validate PLA synthesis | Develop bioassay | Apply MAGE | Introduce export system | Make a bioplastic |
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Aims for the Project
- Engineer strains of E. coli to validate PLA synthesis
- Develop bioassay to screen PLA production
- Apply MAGE to optimize PLA production, guided by FBA
- Introduce type 1 secretion system to export and extract PLA
- Make a bioplastic
Engineer strains of E. coli to validate PLA synthesis
- In order to reproduce the results of the Lee group we needed to insert the two heterologous genes.
- Clostridum propionicum propionate CoA transferase (denoted PCT)
- Pseudomonas resinovorans polyhydroxyalkanoate synthase (denoted PHA)
Here is a schematic of what the entire construct looks like with both promoter, genes and terminators Each fragment was amplified using PCR (according to the protocol here!!!!!!!!!!), to give homology to adjacent fragments. Using gibson assembly our plan was to assemble all 10 fragments into one construct.