Difference between revisions of "Part:BBa K1065001"

Figure 1 Effect of EFE on cell growth. Cell density was measured at different time points to determine the effect of EFE expression. Cells were grown at 37 °C in LB until it was reached an OD of 0.6. The cells were then splitted in four samples of equal volume. Two samples were then induced with 5 mM Arabinose. Induced samples show a slowed growth rate, as espected (5mM arabinose is a strong induction that causes stress on cells). However, cell growth is not completely inhibited so EFE is not highly toxic

Figure 2 Ethylene detection through Micro GC. Cells were grown until O.D.600 reached 0.5. The cells were then splitted in two samples of equal volume (3 ml) and putted into an hermetically closed vial with a septum with a rubber cap. One of the two sample was induced with 5 mM Arabinose. The vials were left in the thermoshaker for 4 hours. After that, the vials were connected to a micro GC and a measure was taken. Panel A: induced sample (green and red curve) showed a characteristic peak corresponding to Ethylene. On the other hand, the not induced sample (blue curve) didn't show the peak. Panel B: picture of the vial connected to the micro GC.

Figure 3 Kinetic assay for Ethylene production. Cells were grown until an O.D. of 0.5 - 0.8 and then connected to the micro GC, while in agitation on a thermoblock at 37 °C for the entire duration of the experiment. Every 45/60 mins a meausure was taken for a total of about 8 hours. Samples were induced at two differents O.D.600 and this had big effect on the amount of ethylene produced. However, it seems that the Ethylene concentration in the air space reached saturation after only two hours. The red dashed line indicates the amount of ethylene detected with a culture left in the thermoshaker for the all duration of the experiment and subjected to only one measurement. As expected an higher value of ethylene was measured due to the minimal gas loss with this approach.

Figure 4 Acceleration of fruit ripening. Our system was exploited for the acceleration of fruit ripening. We designed an ermetically closed jam jar with a rubber hose connector. These jars contained our test-fruit and each one was connected to a flask. The flasks contained 300 ml of induced (or not) culture when his O.D.600 reached 0.8. The flasks contained also a stirrer.The cultures were maintained at 37 °C using a laboratory heating plate connected to a digital thermometer immersed in the culture. For four to six days, every morning the culture in the flasks was substituted with a new induced (or not) colture. Furthermore, canonical jam jars (i.e.: with no connector) were adopted to contain the negative control fruit samples (panel A). We tested many type of fruit, using different time of treatment. In this example (panel B) the tomatoe putted in the jar that was connected to an induced culture, shows a more advance stage of maturation after 4 days of threatment.