Team:KAIT JAPAN/Design

Design

Overview

The objective of our project is to let the E.coli respond to the signal of the cell, and secrete the necessary amount of H₂S synthase (CTH) for the state of suspended animation to lower the metabolism. The secretion of CTH is regulated by RhlR . When the cells to be preserved become the state of suspended animation, the secretion of CTH from E.coli will stop, and most of the E.coli will also be suicided.

Fig.1 Overall Picture

From the Yeast (eukaryotic) side, RhlR used by Pseudomonas aeruginosa for quorum sensing is secreted. In this case, using a secretion vector, it prevents cells from expressing proteins other than the objective.

Fig.2 Securate of RhlR

RhlR to which CPP binds invades E. coli and binds to the promoter. The CTH of interest is placed downstream of the promoter activated by RhlR. By doing so, CTH is expressed in the presence of RhlR. When CTH is secreted from E. coli, a signal protein called HlyA and its corresponding membrane protein are used.

Fig.3 Securate of CTH

As with CPP-bound CTH, it penetrates into Yeast and reacts with L-lysine to produce H 2 S, just as it entered E. coli.

Fig.4 CTH penetrate to Eucaliotic Cells

As a result, H2S acts on mitochondria and Yeast falls into Suspended animation.

Fig.4 Eucaliotic Cells become state of suspended animation