Team:Qdai/Engineering

Engineering

Engineering Success

We have not been able to perform experiments because access to the laboratory was almost impossible until recently due to COVID-19. Therefore, we assume that the experiments will be carried out when access to the laboratory is available.

We plan to introduce three genes, ppk, relA, and phnCDE. Based on our previous research, they may not lead to the accumulation of polyphosphate as desired.

ppk

ppk uses ATP to synthesize polyphosphate. If we introduce ppk into a plasmid with high expression, ATP, which is necessary to maintain the organism, is only used up for the synthesis of polyphosphate, and the growth of E. coli itself may be significantly inhibited. It will be possible to estimate the impact of ppk on growth by changing the expression levels of plasmids containing only ppk and observing their variation.

phnCDE

phnCDE is a transporter of phosphoric acid, but since the lyase is located next to the transporter, it is considered to transport the organic phosphorus mainly. Therefore, if only orthophosphate is added when creating a phosphorus-rich environment, it may not be very effective. The effect of phnCDE should be properly reflected by adding organophosphorus in the environment.

Dual Plasmid System

By introducing ppk and relA into different plasmids using promoters pLlacO1 and pLtetO1 to create a dual plasmid system. The advantage of the system is that induction can be performed separately, allowing us to compare results to the modeling equations, making it easier to improve them.