Experimental Design
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In order to appropriate enhanced IPP and subsequently latex precursor production, we took advantage of E. coli's endogenous MEP pathway, and implemented an HMG-CoA reductase pathway for enhanced IPP production. Both pathways result in the production of IPP and its isomer, DMAPP. However, the HMG-CoA pathway is exogenous to E. coli and was needed for enhanced production of IPP. While the MEP pathway converts pyruvate and G3P into IPP/DMAPP through a seven step process, the HMG-CoA (MVA) pathway converts acetyl CoA into IPP/DMAPP through a six step process. Both pathways employ the use of metabolic products produced and used by glycolysis and cellular respiration (calvin cycle); hence enhanced expression of these pathways is expected to affect cellular growth and development.
Genetic Constructs
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