Team:XHD-Wuhan-China/Engineering
Engineering
To achieve the purpose of further dissolving phosphate, we designed the Phosphate dissolving Plus system.
Phosphate dissolving Plus system
We design an enhanced Phosphate dissolving system (Phosphate dissolving Plus system) which has two plasmids: gcd-pRADK, gabY-pRADK. And we designed gabY gene circuit PGroES-RBS-gabY (Figure 1). In order to realize the function of gabY gene circuit, we constructed the following plasmid gabY-pRADK (Figure 2).
Figure 1. Constitution of PGroES-RBS-gabY gene circuits.
Figure 2. Design of Phosphate dissolving Plus plasmid (gabY-pRADK).
Usage and Biology
We verified the length of the recombinant plasmid to ensure the success of the recombinant plasmid through enzyme digestion (Figure 3). Theoretically, after transforming gabY-pRADK to DR containing gcd-pRADK, the effective concentration of the holoenzyme formed by GDH and PQQ coenzymes will increase, and the catalytic efficiency will be higher. We cultured DR containing gcd-pRADK, gabY-pRADK; DR containing gcd-pRADK; and wild-type DR in TGY liquid medium, and measured pH every 1 hour. The results are shown in the (Figure 4). In addition, DR containing gcd-pRADK, gabY-pRADK; DR containing gcd-pRADK; and wild-type DR were cultured in PKO solid medium, and the results of the phosphate ring were observed, as shown in the (Figure 5).
Figure 3. ElectropHoresis of plasmid gabY-pRADK with enzyme digestions.
Figure 4. Changes in pH value of TGY liquid medium culturing DR R1, DR containing gcd-pRADK and DR containing pRADK2 respectively.
Figure 5. Phosphate ring in PKO solid medium culturing DR R1, DR containing gcd-pRADK and DR containing pRADK2 respectively. Left: DR R1; Middle: DR containing gcd-pRADK; Right: DR containing pRADK2.
Conclusion
We constructed a phosphate dissolving Plus system in Deinococcus radiodurans and successfully verified the reliability of the phosphate dissolving Plus system. At the same time, we also compare the differences of different systems and those can improve the Martian soil in a targeted manner. Our project can provide new solutions to improve the Martian soil and increase the possibility of growing crops on Mars.
