Team:LCG-UNAM-Mexico/Experiments

  The main goal of the experimental work is to test our system's performance. We expect to achieve this by assembling two devices which will carry the system components. Once we have them we will proceed to test the system's behavior.

  The first step is to assemble both devices. One of them includes LuxR, cI+ALV and aiiA; the other one carries the cI promoter, RcnR operator and the RcnA gene. We will use classic techniques (PCR amplification, restriction digest, cohesive end ligation, heat-shock transformation), to assemble all parts and transform the final receptor cell. In our approach we considered to use pJet1.2, (blunt-ending plasmid, high copy number), as an intermediate to clone the amplified parts. The final plasmids will be pRK415 and pBBR1MCS-5 respectively.

  As a second step, we will proceed to test our system's behavior. Once we get YohM- cells(YohM was the name that the ORF coding RcnA used to have) with the RcnA device (BBa_K119009), we will measure changes in the medium's resistivity in order to get the nickel's extrusion and internalization parameters. Our second device (BBa_K119010/BBa_K119011), will let us control our system. We expect to be manipulating the Nickel's concentration in the medium which will be shown through the measurements and the sound emision. Let bacteria sing!!!